Patent application title: COMPOSITIONS AND METHODS FOR TREATING AND DIAGNOSING IRRITABLE BOWEL SYNDROME
Inventors:
Pankaj Pasricha (Houston, TX, US)
Mohan Shenoy (Galveston, TX, US)
John Winston (League City, TX, US)
Assignees:
The Board of Regents of the University of Texas System
IPC8 Class: AC40B3004FI
USPC Class:
506 9
Class name: Combinatorial chemistry technology: method, library, apparatus method of screening a library by measuring the ability to specifically bind a target molecule (e.g., antibody-antigen binding, receptor-ligand binding, etc.)
Publication date: 2012-05-10
Patent application number: 20120115746
Abstract:
Compositions and methods for diagnosing and treating CVH and
CVH-associated disorders are disclosed. Genes differentially expressed in
CVH tissues relative to normal tissues are identified. The genes and the
gene products (i.e., the polynucleotides transcribed from and
polypeptides encoded by the genes) can be used as markers of CVH. The
genes and the gene products can also be used to screen agents that
modulate the gene expression or the activities of the gene products.Claims:
1. A method for detecting CVH in a subject, said method comprising: (a)
contacting a biological sample with an agent that specifically binds to a
polypeptide encoded by a gene listed in Tables 3-8 or a homolog thereof;
(b) determining a level of binding of the agent to the polypeptide; (c)
comparing the level of binding of the agent in the biological sample to a
level of binding of the agent in a normal control sample; and (d)
producing a diagnosis based on a result from step (c).
2. The method of claim 1, wherein said polypeptide comprises an amino acid sequence recited in any one of SEQ ID NOS:34-66.
3. The method of claim 1, wherein the agent is an antibody directed against the polypeptide.
4. A method for detecting CVH in a subject, said method comprising the steps of: (a) determining a level of a transcribed polynucleotide in a biological sample obtained from the subject, wherein the transcribed polynucleotide is transcribed from a gene listed in Tables 3-8 or homolog thereof; (b) comparing the level of the transcribed polynucleotide in the biological sample to a normal level of the transcribed polynucleotide; and (c) producing a diagnosis based on a result from step (b).
5. The method of claim 4, wherein said transcribed polynucleotide comprises a nucleic acid sequence recited in any one of SEQ ID NOS:1-33, or a complement of any of the foregoing nucleic acid sequences.
6. The method of claim 4, wherein the transcribed polynucleotide is an mRNA.
7. A method for detecting CVH in a subject, said method comprising the steps of: (a) determining an expression pattern of polypeptides in a biological sample, said polypeptides are encoded by the genes listed in Tables 3-8 or homologs thereof; (b) comparing the expression pattern of polypeptides obtained in (a) to a normal expression pattern of polypeptides; and (c) producing a diagnosis based on a result from step (b).
8. The method of claim 7, wherein the polypeptides comprise polypeptide sequences recited in SEQ ID NOS:34-66.
9. The method of claim 7, wherein the expression pattern of polypeptides in the biological sample is determined using antibodies directed against the polypeptides.
10. The method of claim 7, wherein the expression pattern of polypeptides is determined by measuring expression levels of at least three polypeptides.
11. A method for detecting CVH in a subject, said method comprising the steps of: (a) determining an expression pattern of mRNAs in a biological sample, said mRNAs are transcribed from the genes listed in Tables 3-8 or homologs thereof; (b) comparing the expression pattern of mRNAs obtained in (a) to a normal expression pattern of mRNAs; and (c) producing a diagnosis based on a result from step (b).
12. The method of claim 11, wherein said mRNAs comprise polynucleotide sequences recited in SEQ ID NOS:1-33.
13. The method of claim 11, wherein the expression pattern of mRNAs is determined by measuring expression levels of at least three mRNAs.
14.-27. (canceled)
24. A biochip comprising at least one of: (a) a polynucleotide comprising a sequence that hybridizes to a gene listed in Tables 3-8 or a homolog thereof; (b) a polypeptide comprising at least a portion of a sequence encoded by a gene listed in Tables 3-8.
25. The biochip of claim 24, wherein said polynucleotide comprises a sequence that hybridizes to any one of SEQ ID NOS: 1-33, and wherein said polypeptide comprising any one of SEQ ID NOS:34-66.
26. A diagnostic kit for CVH, said kit comprising at least one of: a polynucleotide probe, wherein the probe specifically binds to a polynucleotide transcribed from a gene listed in Tables 3-8 or a homolog thereof; and an antibody capable of immunospecific binding to a polypeptide encoded by a gene listed in tables 3-8.
27. The diagnostic kit of claim 26, wherein the probe specifically binds to a transcribed polynucleotide comprising any one of SEQ ID NOS:1-33.
28. The diagnostic kit of claim 26, wherein the antibody specifically binds to a polypeptide comprising any one of SEQ ID NOS:34-66.
29.-31. (canceled)
Description:
[0001] This application claims priority from U.S. Provisional Application
Ser. No. 60/496,716 filed Aug. 21, 2003. The entirety of that provisional
application is incorporated herein by reference.
TECHNICAL FIELD
[0002] The present invention relates generally to the diagnosis and treatment of disorders associated with chromic visceral hypersensitivity (CVH), and in particular irritable bowel syndrome (IBS). The invention also relates to genes associated with CVH, polynucleotides transcribed from these genes and polypeptides encoded by these genes. Such polynucleotides and polypeptides can be used for the diagnosis and treatment of CVH.
BACKGROUND OF THE INVENTION
[0003] Irritable Bowel Syndrome (IBS) is a functional bowel disorder of unknown etiology. A functional disorder refers to a disorder or disease where the primary abnormality is an altered physiological function, rather than an identifiable structural or biochemical cause. IBS is characterized by a group of symptoms including intermittent abdominal pain and discomfort and alterations in bowel habits, such as loose or more frequent bowel movements, diarrhea, and/or constipation that occur in the absence of detectable ongoing organic disease.
[0004] IBS affects approximately 10-20% of the general population. It is the most common disease diagnosed by gastroenterologists and one of the most common disorders seen by primary care physicians. IBS is understood as a multi-faceted disorder. In people with IBS, symptoms result from what appears to be a disturbance in the interaction between the gut or intestines, the brain, and the autonomic nervous system that alters regulation of bowel motility (motor function) or sensory function.
[0005] Human studies demonstrate that IBS is associated with a state of chronic visceral hypersensitivity (CVH) suggesting that processing of visceral sensory information is altered. However, little is known about how the afferent nervous system is changed in this syndrome. A hallmark of TBS is increased visceral hypersensitivity, but the molecular changes underlying the development and maintenance of chronic visceral hypersensitivity in IBS are not known. Current medical treatments for IBS primarily target peripheral symptoms rather than the underlying causes, and therapeutic gains from drug treatments are usually modest and the placebo responses are high (Mertz et al., Gastroenterology, 109:40-52, 1995). Defining the underlying neurological and molecular defects is therefore important to the design of more successful therapeutic strategies. Moreover, there is a need in the art for improved methods for screening, diagnosing, and treating MS and other CVH-related disorders.
[0006] CNI-1493 is a MAPK and TNF inhibitor with anti-inflammatory and possible analgesic actions. CNI-1493 inhibits signal transduction pathways by preventing phosphorylation of p38 MAP kinase and JNK, and inhibits production of the proinflammatory cytokines such as TNF-alpha, IL-1, MIP-1 alpha, and MIP-1 beta. In animal models, CNI-1493 has shown protective activity against a wide variety of conditions, ranging from stroke to inflammatory bowel disease. However, CNI-1493 has never been tested for its anti-nociceptive activity in the absence of inflammation. Recently, an animal model of chronic visceral hypersensitivity was created using mechanical and chemical irritation of the colon of neonatal rats (Al-Chaer et al., Gastroenterology, 119:1276-1285, 2000). The animal model provides an ideal platform for studying IBS, validating the neurogenic components of functional abdominal pain, and testing agents that may reduce visceral hypersensitivity.
SUMMARY OF THE INVENTION
[0007] One aspect of the present invention relates to the treatment for disorders associated with CVH using guanylhydrazone. In one embodiment, the present invention provides a treatment for IBS using CNI1493.
[0008] Another aspect of the present invention relates to CVH-related genes (CVHGs) and the gene products, which include the polynucleotides transcribed from the CVHGs (CHVPNs) and the polypeptides encoded by the CVHGs (CHVPPs).
[0009] In one embodiment, the present invention provides methods for diagnosing and monitoring CVH and CVH-related disorders by comparing the expression levels of one or more CVHGs at the nucleotide or protein level in biological samples from a subject to control samples.
[0010] In another embodiment, the present invention provides pharmaceutical compositions for the treatment of CVH and CVH-related disorders. The pharmaceutical compositions comprise a pharmaceutically acceptable carrier and at least one of the following: (1) a CVHG product; (2) an agent that modulates an activity of a CVHG product; and (3) an agent that modulates the expression of a CVHG.
[0011] In another embodiment, the present invention provides methods for treating CVH and CVH-related disorders in a patient with the pharmaceutical compositions described above. The patient may be afflicted with CVH, in which case the methods provide treatment for the disease. The patient may also be considered at risk for CVH, in which case the methods provide prevention for disease development.
[0012] In another embodiment, the present invention provides methods for screening anti-CVH agents based on the agents' interaction with CVHPPs, or the agents' effect on the activity or expression of CVHPPs.
[0013] In another embodiment, the present invention provides biochips for diagnosing CVH and CHH-related disorders, and for screening agents that inhibit CVH. The biochips comprise at least one of the following (1) a CVHPP or its variant, (2) a portion of a CVHPP or its variant (3) a CVHPN or its variant, and (4) a portion of a CVHPN or its variant.
[0014] In another embodiment, the present invention provides a kit for diagnosing CVH and CVH-related disorders. The kit comprises at least one of the following (1) polynucleotide probe that specifically hybridizes to a CVHPN, and (2) an antibody capable of specific binding to a CVHPP.
BRIEF DESCRIPTION OF FIGURES
[0015] FIG. 1. Increased sensitivity to CRD in adult rats treated with acetic acid on P10 (n=8). Data was analyzed by two-way repeated measures ANOVA with distention pressure as the repeated factor and P10 treatment as a between group factor. There was a significant effect of P10 treatment (F 1, 12.98) p<0.003, of distention pressure (F 7, 89.9) p<0.001, and there was a significant interaction between distention pressure and P10 treatment (F 7, 4.04) p<0.001. Means were compared with a Tukey test. Significant differences between acetic acid treated and controls were found at distention pressures of 30 (p=0.004), 40 (p<0.001), 50 (p<0.001), 60 (p=0.001) and 70 (p=0.035) mm Hg.
[0016] FIG. 2. Effect of CNI-1493 on the response of sensitized rats to graded CRD (n=8). Data was analyzed by two-way repeated measures ANOVA with distention pressure as the repeated factor and drug treatment as a between group factor. There was a significant effect of CNI1493 treatment (F 1, 16.96) p=0.001, of distention pressure (F 7, 28.55) p<0.001, but there was no significant interaction between distention pressure and CNI1493 treatment (F 7, 1.94) p=0.071. Means were compared with a Tukey test. Significant differences between CNI-1493 treated and controls were found at distention pressures of 20 (P=0.001), 30 (p=0.003), 40 (p<0.001), 50 (p=0.001), 60 (p<0.001) and 70 (p=0.015) and 80 (p=0.007) mm Hg.
[0017] FIG. 3. Colon histology and MPO activity. H&E stained colon sections from control, vehicle (A); control, cni-1493 (B); sensitized, vehicle (C); and sensitized, CNI1493 (D). Histogram showing MPO activity in colons (E).
DETAILED DESCRIPTION OF THE INVENTION
[0018] The preferred embodiments of the invention are described below. Unless specifically noted, it is intended that the words and phrases in the specification and claims be given the ordinary and accustomed meaning to those of ordinary skill in the applicable art or arts. If any other meaning is intended, the specification will specifically state that a special meaning is being applied to a word or phrase.
[0019] It is further intended that the inventions not be limited only to the specific structure, material or acts that are described in the preferred embodiments, but in addition, include any and all structures, materials or acts that perform the claimed function, along with any and all known or later-developed equivalent structures, materials or acts for performing the claimed function.
[0020] Further examples exist throughout the disclosure, and it is not applicant's intention to exclude from the scope of his invention the use of structures, materials, methods, or acts that are not expressly identified in the specification, but nonetheless are capable of performing a claimed function.
[0021] The present invention is generally directed to compositions and methods for the diagnosis, treatment, and prevention of CVH and CVH-related disorders; and to the identification of novel therapeutic agents for CVH and CVH-related disorders. The present invention is based on the finding that guanylhydrazone is capable of ameliorating CVH in a rat model of IBS and the discovery of transcribed polynucleotides that are differentially expressed in the colon tissue of rats with chemically induced CVH relative to control animals.
DEFINITIONS AND TERMS
[0022] To facilitate an understanding of the present invention, a number of terms and phrases are defined below:
[0023] As used herein, the terms "a differentially expressed gene" refer to a gene that meets all of the following criteria during an Affymetrix microarray analysis: (1) the average expression of the gene shows a fold change of two or greater compared with controls and (2) significant changes in gene expression were detected by analyzing signal intensity values by two-way ANOVA with 99% confidence. The differentially expressed genes identified in the colon tissue samples of CVH and CNI1493-treated rats are designated as CVH-related genes (CVHGs). CVHGs generally refer to the genes listed in Tables 3-8.
[0024] As used herein, the terms "CVH-related polynucleotide (CVHPN)" and "CVHG polynucleotide" are used interchangeably. The terms include a transcribed polynucleotide (e.g., DNA, cDNA or mRNA) that comprises one of the CVHG sequences or a portion thereof.
[0025] As used herein, the terms "CVH-related polypeptide (CVHPP)" and "CVHG protein" are used interchangeably. The terms include polypeptides encoded by an CVHG, an CVHPN, or a portion of an CVHG or CVHPN.
[0026] As used herein, a "CVHG product" includes a nucleic acid sequence and an amino acid sequence (e.g., a polynucleotide or polypeptide) generated when an CVHG is transcribed and/or translated. Specifically, CVHG products include CVHPNs and CVHPPs.
[0027] As used herein, a "variant of a polynucleotide" includes a polynucleotide that differs from the original polynucleotide by one or more substitutions, additions, deletions and/or insertions such that the activity of the encoded polypeptide is not substantially changed (e.g., the activity may be diminished or enhanced, by less than 50%, and preferably less than 20%) relative to the polypeptide encoded by the original polynucleotide.
[0028] A variant of a polynucleotide also includes polynucleotides that are capable of hybridizing under reduced stringency conditions, more preferably stringent conditions, and most preferably highly stringent conditions to the original polynucleotide (or a complementary sequence). Examples of conditions of different stringency are listed in Table 2.
[0029] It will be appreciated by those of ordinary skill in the art that, as a result of the degeneracy of the genetic code, there are many nucleotide sequences that encode a polypeptide as described herein. Some of these polynucleotides bear minimal homology to the nucleotide sequence of any native gene. Nonetheless, polynucleotides that vary due to differences in codon usage are specifically contemplated by the present invention.
[0030] As used herein, a "variant of a polypeptide" is a polypeptide that differs from a native polypeptide in one or more substitutions, deletions, additions and/or insertions, such that the bioactivity or immunogenicity of the native polypeptide is not substantially diminished. In other words, the bioactivity of a variant polypeptide or the ability of a variant polypeptide to react with antigen-specific antisera may be enhanced or diminished by less than 50%, and preferably less than 20%, relative to the native polypeptide. Variant polypeptides include those in which one or more portions, such as an N-terminal leader sequence or transmembrane domain, have been removed. Other preferred variants include variants in which a small portion (e.g., 1-30 amino acids, preferably 5-15 amino acids) has been removed from the N- and/or C-terminal of the mature protein.
[0031] Modifications and changes can be made in the structure of a polypeptide of the present invention and still obtain a molecule having biological activity and/or immunogenic properties. Because it is the interactive capacity and nature of a polypeptide that defines that polypeptide's biological activity, certain amino acid sequence substitutions can be made in a polypeptide sequence (or, of course, its underlying DNA coding sequence) and nevertheless obtain a polypeptide with like properties.
[0032] In making such changes, the hydropathic index of amino acids can be considered. The importance of the hydropathic amino acid index in conferring interactive biologic function on a polypeptide is generally understood in the art. It is believed that the relative hydropathic character of the amino acid residue determines the secondary and tertiary structure of the resultant polypeptide, which in turn defines the interaction of the polypeptide with other molecules, such as enzymes, substrates, receptors, antibodies, antigens, and the like. It is known in the art that an amino acid can be substituted by another amino acid having a similar hydropathic index and still obtain a functionally equivalent polypeptide. In such changes, the substitution of amino acids whose hydropathic indices are within +/-2 is preferred, those that are within +/-1 are particularly preferred, and those within +/-0.5 are even more particularly preferred.
[0033] Substitution of like amino acids can also be made on the basis of hydrophilicity, particularly where the biological functional equivalent polypeptide or polypeptide fragment, is intended for use in immunological embodiments. U.S. Pat. No. 4,554,101, incorporated hereinafter by reference, states that the greatest local average hydrophilicity of a polypeptide, as governed by the hydrophilicity of its adjacent amino acids, correlates with its immunogenicity and antigenicity, i.e. with a biological property of the polypeptide.
[0034] As detailed in U.S. Pat. No. 4,554,101, the following hydrophilicity values have been assigned to amino acid residues: arginine (+3.0); lysine (+3.0); aspartate (+3.0±1); glutamate (+3.0±1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); proline (-0.5±1); threonine (-0.4); alanine (-0.5); histidine (-0.5); cysteine (-1.0); methionine (-1.3); valine (-1.5); leucine (-1.8); isoleucine (-1.8); tyrosine (-2.3); phenylalanine (-2.5); tryptophan (-3.4). It is understood that an amino acid can be substituted for another having a similar hydrophilicity value and still obtain a biologically equivalent, and in particular, an immunologically equivalent polypeptide. In such changes, the substitution of amino acids whose hydrophilicity values are within ±2 is preferred, those that are within ±1 are particularly preferred, and those within ±0.5 are even more particularly preferred.
[0035] As outlined above, amino acid substitutions are generally therefore based on the relative similarity of the amino acid side-chain substituents, for example, their hydrophobicity, hydrophilicity, charge, size, and the like. Exemplary substitutions which take various of the foregoing characteristics into consideration are well known to those of skill in the art and include: arginine and lysine; glutamate and aspartate; serine and threonine; glutamine and asparagine; and valine, leucine and isoleucine (See Table 1, below). The present invention thus contemplates functional or biological equivalents of an CVHPP as set forth above.
TABLE-US-00001 TABLE 1 Amino Acid Substitutions Exemplary Residue Original Residue Substitution Ala Gly; Ser Arg Lys Asn Gln; His Asp Glu Cys Ser Gln Asn Glu Asp Gly Ala His Asn; Gln Ile Leu; Val Leu Ile; Val Lys Arg Met Leu; Tyr Ser Thr Thr Ser Trp Tyr Tyr Trp; Phe Val Ile; Leu
[0036] A variant may also, or alternatively, contain nonconservative changes. In a preferred embodiment, variant polypeptides differ from a native sequence by substitution, deletion or addition of five amino acids or fewer. Variants may also (or alternatively) be modified by, for example, the deletion or addition of amino acids that have minimal influence on the immunogenicity, secondary structure, tertiary structure, and hydropathic nature of the polypeptide.
[0037] Polypeptide variants preferably exhibit at least about 70%, more preferably at least about 90% and most preferably at least about 95% sequence homology to the original polypeptide.
[0038] A polypeptide variant also includes a polypeptide that is modified from the original polypeptide by either natural processes, such as post-translational processing, or by chemical modification techniques which are well known in the art. Modifications can occur anywhere in a polypeptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given polypeptide. Also, a given polypeptide may contain many types of modifications. Polypeptides may be branched, for example, as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched, and branched cyclic polypeptides may result from post-translation natural processes or may be made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a fluorophore or a chromophore, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination.
[0039] As used herein, a "biologically active portion" of a CVHPP includes a fragment of a CVHPP comprising amino acid sequences sufficiently homologous to or derived from the amino acid sequence of the CVHPP, which includes fewer amino acids than the full length CVHPP, and exhibits at least one activity of the CVHPP. Typically, a biologically active portion of a CVHPP comprises a domain or motif with at least one activity of the CVHPP. A biologically active portion of a CVHPP can be a polypeptide which is, for example, 10, 25, 50, 100, 200 or more amino acids in length. Biologically active portions of a CVHPP can be used as targets for developing agents which modulate a CVHPP-mediated activity.
[0040] As used herein, an "immunogenic portion," an "antigen," an "immunogen," or an "epitope" of a CVHPP includes a fragment of a CVHPP comprising an amino acid sequence sufficiently homologous to, or derived from, the amino acid sequence of the CVHPP, which includes fewer amino acids than the full length CVHPP and can be used to induce an anti-CVHPP humoral and/or cellular immune response.
[0041] As used herein, the term "modulation" includes, in its various grammatical forms (e.g., "modulated", "modulation", "modulating", etc.), up-regulation, induction, stimulation, potentiation, and/or relief of inhibition, as well as inhibition and/or down-regulation or suppression.
[0042] As used herein, the term "control sequences" or "regulatory sequences" refers to DNA sequences necessary for the expression of an operably linked coding sequence in a particular host organism. The term "control/regulatory sequence" is intended to include promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Control/regulatory sequences include those which direct constitutive expression of a nucleotide sequence in many types of host cells and those which direct expression of the nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences).
[0043] A nucleic acid sequence is "operably linked" to another nucleic acid sequence when the former is placed into a functional relationship with the latter. For example, a DNA for a presequence or secretory leader peptide is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide; a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation. Generally, "operably linked" means that the DNA sequences being linked are contiguous and, in the case of a secretory leader, contiguous and in reading phase. However, enhancers do not have to be contiguous. Linking is accomplished by ligation at convenient restriction sites. If such sites do not exist, synthetic oligonucleotide adaptors or linkers are used in accordance with conventional practice.
[0044] As used herein, the "stringency" of a hybridization reaction refers to the difficulty with which any two nucleic acid molecules will hybridize to one another. The present invention also includes polynucleotides capable of hybridizing under reduced stringency conditions, more preferably stringent conditions, and most preferably highly stringent conditions, to polynucleotides described herein. Examples of stringency conditions are shown in Table 2 below: highly stringent conditions are those that are at least as stringent as, for example, conditions A-F; stringent conditions are at least as stringent as, for example, conditions G-L; and reduced stringency conditions are at least as stringent as, for example, conditions M-R.
TABLE-US-00002 TABLE 2 Stringency Condition Poly- Stringency nucleotide Hybrid Hybridization Wash Temp. Condition Hybrid Length (bp)1 Temperature and BufferH and BufferH A DNA:DNA >50 65° C.; 1xSSC -or- 65° C.; 42° C.; 1xSSC, 50% formamide 0.3xSSC B DNA:DNA <50 TB*; 1xSSC TB*; 1xSSC C DNA:RNA >50 67° C.; 1xSSC -or- 67° C.; 45° C.; 1xSSC, 50% formamide 0.3xSSC D DNA:RNA <50 TD*; 1xSSC TD*; 1xSSC E RNA:RNA >50 70° C.; 1xSSC -or- 70° C.; 50° C.; 1xSSC, 50% formamide 0.3xSSC F RNA:RNA <50 TF*; 1xSSC TF*; 1xSSC G DNA:DNA >50 65° C.; 4xSSC -or- 65° C.; 42° C.; 4xSSC, 50% formamide 1xSSC H DNA:DNA <50 TH*; 4xSSC TH*; 4xSSC I DNA:RNA >50 67° C.; 4xSSC -or- 67° C.; 45° C.; 4xSSC, 50% formamide 1xSSC J DNA:RNA <50 TJ*; 4xSSC TJ*; 4xSSC K RNA:RNA >50 70° C.; 4xSSC -or- 67° C.; 50° C.; 4xSSC, 50% formamide 1xSSC L RNA:RNA <50 TL*; 2xSSC TL*; 2xSSC M DNA:DNA >50 50° C.; 4xSSC -or- 50° C.; 40° C.; 6xSSC, 50% formamide 2xSSC N DNA:DNA <50 TN*; 6xSSC TN*; 6xSSC O DNA:RNA >50 55° C.; 4xSSC -or- 55° C.; 42° C.; 6xSSC, 50% formamide 2xSSC P DNA:RNA <50 TP*; 6xSSC TP*; 6xSSC Q RNA:RNA >50 60° C.; 4xSSC -or- 60° C.; 45° C.; 6xSSC, 50% formamide 2xSSC R RNA:RNA <50 TR*; 4xSSC TR*; 4xSSC 1The hybrid length is that anticipated for the hybridized region(s) of the hybridizing polynucleotides. When hybridizing a polynucleotide to a target polynucleotide of unknown sequence, the hybrid length is assumed to be that of the hybridizing polynucleotide. When polynucleotides of known sequence are hybridized, the hybrid length can be determined by aligning the sequences of the polynucleotides and identifying the region or regions of optimal sequence complementarity. HSSPE (1xSSPE is 0.15M NaCl, 10 mM NaH2PO4, and 1.25 mM EDTA, pH 7.4) can be substituted for SSC (1xSSC is 0.15M NaCl and 15 mM sodium citrate) in the hybridization and wash buffers; washes are performed for 15 minutes after hybridization is complete. TB* - TR*: The hybridization temperature for hybrids anticipated to be less than 50 base pairs in length should be 5-10° C. less than the melting temperature (Tm) of the hybrid, where Tm is determined according to the following equations. For hybrids less than 18 base pairs in length, Tm(° C.) = 2(# of A + T bases) + 4(# of G + C bases). For hybrids between 18 and 49 base pairs in length, Tm(° C.) = 81.5 + 16.6(log10Na+) + 0.41(% G+C) - (600/N), where N is the number of bases in the hybrid, and Na+ is the concentration of sodium ions in the hybridization buffer (Na+ for 1xSSC = 0.165M).
[0045] As used herein, the terms "immunospecific binding" and "specifically bind to" refer to antibodies that bind to an antigen with a binding affinity or 105 Moles taken either from pre-CVH or from a subject who has not suffered CVH, or from a cell, tissue or sample that is not affected by CVH. Control samples of the present invention are taken from normal samples.
[0046] As used herein, the term "expression pattern" includes expression of a group of genes at RNA or protein level, the quantity or activity of each member of which is correlated with the incidence or risk of incidence of CVH and CVH associated diseases. An expression pattern comprises expression level of 2 or more CVHGs. An expression pattern may also comprise expression level of 2-5, 5-15, 15-35, 35-50, or more than 50 CVHGs.
[0047] As used herein, the terms "treating," "treatment," and "therapy" refer to curative therapy, prophylactic therapy, and preventative therapy.
[0048] Various aspects of the invention are described in further detail in the following subsections. The subsections below describe in more detail the present invention. The use of subsections is not meant to limit the invention; subsections may apply to any aspect of the invention.
[0049] One aspect of the present invention relates to a method for treating CVH and CVH-related disorders with compositions comprising a guanylhydrazone compound. In one embodiment, the CVH-related disorder is IBS and the guanylhydrazone compound is CNI1493.
[0050] Another aspect of the present invention relates to CVH-related genes. Briefly, new born rats were sensitized by infusion of acetic acid into the colon at P10. Control rats received saline. At eight weeks, the animals were divided into four groups: control+vehicle; control+CNI1493; sensitized+vehicle; and sensitized+CNI1493. Colon samples were obtained after CNI1493 treatment (5 mg/kg for four days, the vehicle group received vehicle only). Gene expression patterns in the colon tissue samples and S1 dorsal root ganglia (S1 DRG) were analyzed using Affymetrix rat genome 230A chips (Affymetrix, Santa Clara, Calif.). Single array analysis for each chip was performed by Affymetrix Microarray Suite (MAS) software to produce a detection call, present, absent or marginal and a signal intensity value for each gene that is a relative measure of abundance of the transcript. For comparison of signal intensity values between chips, all chips will be scaled to an average intensity of 500. Genes called "Absent" across all chips and genes without a Fold-change ≧2.0 in at least one of the pairwise comparisons of chips from different treatment groups are excluded. The probe sets with absolute call "Absent" across all chips and Fold-change <2.0 in all of the possible pairwise comparisons, are filtered out. ANOVA was then performed on the filtered data set. Significant changes in gene expression were detected by analyzing signal intensity values by two-way ANOVA with 99% confidence limits. The differentially regulated genes were subjected to cluster analysis to identify genes associated with sensitization and treatment.
[0051] Tables 3 and 4 provides a list of the genes that are differentially expressed in the colon and S1 DRG, respectively, of the sensitized animals, i.e., animals suffering from CVH. Tables 5 and 6 provide a list of the genes that are differentially expressed in the colon and S1 DRG, respectively, in CNI1493-treated animals. Since CNI1493 treatment ameliorates CVH in the sensitized animals, genes differentially regulated by CNI1493 may also be related to the etiology of CVH. Accordingly, genes listed in Tables 3-6 are designated as CVH-related genes (CVHGs).
TABLE-US-00003 TABLE 3 Genes differentially expressed in sensitized colon No. Acce. No. Symbol CTRL-VHL CTRL-CNI IBS-VHL IBS-CNI 73 NM_021769 Sult-n 1.00 0.85 2.98 1.85 57 NM_022531 Des 1.00 1.03 2.55 1.31 14 BF389682 zzN/A 1.00 1.53 2.49 2.04 16 BF419200 Cebpd 1.00 1.03 2.44 2.28 26 BI296437 zzN/A 1.00 1.14 2.43 1.21 25 NM_013002 Pcp4 1.00 0.88 2.29 1.14 55 NM_013122 Igfbp2 1.00 0.75 2.19 0.94 74 NM_024400 Adamts1 1.00 0.90 2.11 1.14 65 AI229029 Tubb3 1.00 0.93 2.05 1.53 43 BI282702 Acta2 1.00 0.95 2.03 0.99 44 BF399310 zzN/A 1.00 0.64 2.01 0.78 64 NM_031970 Hspb1 1.00 1.14 1.90 1.40 12 ILGFBP4 BC019836 1.00 1.41 1.88 2.00 58 BF290193 zzN/A 1.00 0.65 1.84 0.90 36 AA012755 1.00 0.72 1.83 0.79 45 AI103600 zzN/A 1.00 0.57 1.81 0.58 63 AA851939 Fxyd6 1.00 0.95 1.81 1.26 56 AA799832 zzN/A 1.00 0.78 1.80 0.95 50 NM_019904 Lgals1 1.00 0.74 1.79 0.96 37 NM_017148 Csrp1 1.00 0.63 1.72 0.71 35 BI283060 zzN/A 1.00 0.63 1.71 0.69 39 AA800892 zzN/A 1.00 0.57 1.71 0.68 29 BG373779 LOC308709 1.00 0.48 1.68 0.59 2 BI285494 Ifitm31 1.00 3.86 1.66 4.90 47 NM_053770 Argbp2 1.00 0.79 1.66 0.77 34 NM_012893 Actg2 1.00 0.48 1.66 0.56 46 NM_130403 Ppp1r14a 1.00 0.91 1.65 0.99 15 BG663107 zzN/A 1.00 1.06 1.65 1.23 30 D29960 Loc192245 1.00 0.51 1.64 0.54 48 BM386598 zzN/A 1.00 0.90 1.60 0.95 71 AA818342 zzN/A 1.00 0.88 1.58 1.25 5 AW523747 Amigo2 1.00 0.88 1.57 2.10 40 AA817802 zzN/A 1.00 0.55 1.57 0.69 75 BE112887 zzN/A 1.00 1.06 1.55 1.05 33 M23764 Tpm1 1.00 0.60 1.54 0.68 59 AW522471 Exo70 1.00 0.65 1.54 1.00 8 EST 1.00 0.93 1.52 2.14 31 AI044427 zzN/A 1.00 0.59 1.52 0.69 41 BI279044 zzN/A 1.00 0.49 1.51 0.52 9 EST 1.00 1.48 1.51 2.10 28 BI291848 zzN/A 1.00 0.62 1.51 0.81 24 AI411809 zzN/A 1.00 0.89 1.51 1.06 67 NM_053440 Stmn2 1.00 0.79 1.49 1.06 54 BI285456 zzN/A 1.00 0.83 1.49 1.06 32 NM_031549 Tagln 1.00 0.54 1.47 0.64 38 AI177055 zzN/A 1.00 0.79 1.47 0.82 52 BI279661 Wfdc1 1.00 0.66 1.46 0.94 20 AI229240 zzN/A 1.00 0.68 1.46 1.69 1 BI285346 Ppp4r1 1.00 1.34 1.46 1.51 53 NM_134449 Prkcdbp 1.00 0.87 1.45 1.01 49 BF555956 zzN/A 1.00 0.85 1.45 0.91 42 X16262 Myh11 1.00 0.54 1.44 0.60 51 BG666999 Slc25a4 1.00 0.79 1.44 0.94 70 AI177366 Itgb1 1.00 1.00 1.42 1.24 61 AA686007 Parva 1.00 0.80 1.41 0.99 68 BM389644 zzN/A 1.00 0.88 1.41 1.07 18 BG378721 zzN/A 1.00 0.95 1.40 1.21 60 AA893484 Fn1 1.00 0.51 1.40 0.88 27 NM_019304 Dgkb 1.00 0.67 1.31 0.86 7 EST 1.00 1.09 1.31 1.51 19 NM_017131 Casq2 1.00 0.71 1.30 1.19 6 U06434 Scya4 1.00 1.18 1.30 1.76 10 AB043636 Kcnj8 1.00 1.22 1.29 1.65 11 EST 1.00 0.82 1.29 1.55 66 AF081582 Evt1 1.00 0.85 1.29 1.03 3 NM_024145 Fgr 1.00 1.03 1.26 1.70 72 BI296312 zzN/A 1.00 0.80 1.26 1.04 13 AA859496 Gch 1.00 0.93 1.24 1.34 22 BF394235 zzN/A 1.00 0.55 1.23 0.99 69 BF551377 zzN/A 1.00 0.81 1.21 0.99 62 AI179984 Dmrs91 1.00 0.49 1.21 0.70 21 NM_133605 Camk2g 1.00 0.77 1.18 1.01 17 U27518 LOC286989 1.00 0.50 1.11 1.19 4 NM_022688 Porf1 1.00 0.80 1.08 1.42 23 AW254190 1.00 0.66 1.05 1.03 87 BM386844 zzN/A 1.00 1.40 1.00 0.84 94 BF390024 Ncor1 1.00 1.34 0.93 0.89 92 BE098713 zzN/A 1.00 1.11 0.87 0.82 86 BF404414 zzN/A 1.00 1.19 0.85 0.80 82 AA945828 zzN/A 1.00 1.54 0.84 1.07 88 AA849756 zzN/A 1.00 0.86 0.84 0.66 83 BM392373 Ceacam1 1.00 1.38 0.82 0.87 78 BM385170 zzN/A 1.00 1.30 0.81 1.11 77 BF411331 zzN/A 1.00 1.22 0.80 1.04 104 AI177513 zzN/A 1.00 0.90 0.80 0.70 107 AA964600 zzN/A 1.00 0.83 0.79 0.68 85 BI295141 zzN/A 1.00 1.07 0.78 0.82 108 AI227627 Cd9 1.00 0.74 0.77 0.61 90 NM_031762 Cdkn1b 1.00 1.02 0.74 0.45 89 AI180286 zzN/A 1.00 0.90 0.74 0.70 91 AI105205 Ctl1 1.00 0.93 0.73 0.57 103 BG380281 zzN/A 1.00 0.72 0.72 0.64 101 BI301490 zzN/A 1.00 0.68 0.69 0.66 113 BM384203 zzN/A 1.00 0.90 0.69 0.66 114 BG373555 zzN/A 1.00 0.98 0.68 0.74 105 BI288816 zzN/A 1.00 0.65 0.68 0.48 109 AA894262 zzN/A 1.00 1.07 0.68 0.59 106 AI228548 zzN/A 1.00 0.62 0.67 0.30 76 AW433971 Mvk 1.00 1.12 0.67 0.98 80 AA800750 zzN/A 1.00 1.13 0.65 0.92 84 AI171229 zzN/A 1.00 1.23 0.65 0.78 112 AI407835 Add3 1.00 0.74 0.65 0.65 97 NM_023989 LOC78973 1.00 0.84 0.65 0.70 81 AA893602 zzN/A 1.00 1.13 0.65 0.93 98 AA943165 zzN/A 1.00 0.71 0.64 0.52 93 NM_031347 Ppargc1 1.00 1.17 0.62 0.52 79 BE102350 zzN/A 1.00 1.64 0.61 1.18 110 AI408598 zzN/A 1.00 0.85 0.61 0.53 111 AI175820 zzN/A 1.00 0.64 0.61 0.57 99 AI179665 1.00 0.77 0.59 0.64 100 BI296591 zzN/A 1.00 0.53 0.54 0.55 102 AF189724 Cxcl12 1.00 0.73 0.50 0.61 96 BF417032 zzN/A 1.00 0.87 0.49 0.57 95 M58040 Tfrc 1.00 0.63 0.45 0.42
TABLE-US-00004 TABLE 4 Genes differentially expressed in sensitized S1 DRG No. Identifier Acce. No. CTRL-VHL CTRL-CNI IBS-VHL IBS-CNI 11 1376554_at BE121079 1 1.74 2.23 2.01 23 1369233_at AF196965 1 1.45 2.09 1.71 4 1374620_at BM392373 1 1.43 1.87 2.41 6 1390403_at BE108405 1 1.39 1.83 1.86 8 1379272_at AA963084 1 1.08 1.83 1.70 24 1369157_at NM_017229 1 1.05 1.78 1.09 19 1382915_at AI237079 1 1.26 1.76 1.38 25 1374802_at AI010721 1 0.90 1.72 0.96 5 1389222_at BI282847 1 1.30 1.69 1.66 22 1368379_at NM_054001 1 1.31 1.68 1.66 7 1368678_at X67108 1 1.38 1.66 1.72 10 1386218_at AI639301 1 1.35 1.65 1.58 26 1370301_at U65656 1 1.17 1.64 1.02 17 1389099_at AI600184 1 1.50 1.64 1.50 18 1383159_at AW434445 1 1.40 1.64 1.58 13 1384217_at BI276341 1 1.24 1.59 1.67 21 1389713_at AI602851 1 1.17 1.58 1.45 12 1372620_at AI008642 1 1.32 1.57 1.94 78 1385386_at BI302745 1 0.79 0.09 0.31 28 1387658_at U93849 1 1.60 0.43 0.82 47 1368703_at NM_053326 1 0.65 0.44 0.70 62 1377606_at AI500913 1 0.49 0.44 0.36 50 1375606_at AI235906 1 0.75 0.46 0.43 49 1369255_at NM_013123 1 0.80 0.46 0.58 40 1386999_at BG380730 1 0.78 0.47 0.68 42 1388589_at BG381046 1 0.74 0.47 0.61 41 1370666_at AF201839 1 0.66 0.49 0.44 52 1389864_at BF405086 1 0.69 0.50 0.58 72 1374283_at BF419505 1 0.82 0.51 0.59 34 1375469_at BE111847 1 0.67 0.52 0.67 38 1374002_at AI045904 1 0.71 0.55 0.79 65 1394114_at AA799434 1 0.67 0.56 0.63 67 1388101_at AF389425 1 0.73 0.56 0.71 57 1368279_at NM_053718 1 0.78 0.56 0.39 45 1376350_at BF396151 1 0.90 0.57 0.75 74 1376122_at BF401577 1 0.77 0.57 0.44 36 1369036_at NM_019309 1 0.72 0.58 0.65 66 1367814_at M14137 1 0.67 0.59 0.67 71 1372177_at AI180033 1 0.63 0.60 0.61 70 1373981_at BI299720 1 0.62 0.60 0.55 35 1383468_at BM958512 1 0.66 0.62 0.67 46 1376931_at BG380736 1 0.73 0.62 0.75 75 1371281_at M37568 1 0.97 0.63 0.39 44 1387818_at NM_053736 1 0.87 0.63 0.75 51 1369048_at NM_017289 1 0.71 0.64 0.56 37 1374084_at BE119993 1 0.68 0.64 0.59 30 1387327_at NM_133318 1 1.25 0.65 0.66 39 1373031_at BI275757 1 0.83 0.65 0.77 32 1375448_at BM391628 1 0.90 0.65 0.74 88 1369884_at NM_022182 1 0.90 0.66 0.54 59 1375294_at BF415950 1 0.75 0.66 0.42 90 1387404_at NM_078620 1 1.02 0.66 0.50 69 1386907_at NM_012949 1 0.75 0.67 0.67 63 1392500_at AA957990 1 0.59 0.67 0.58 68 1369000_at NM_021589 1 0.75 0.67 0.67
TABLE-US-00005 TABLE 5 Genes differentially expressed in CNI1493-treated colon No. Identifier Acce. No. Description 1 1370531_at U69550 phospholipase D gene 1 2 1369708_at NM_031017 cAMP response element binding protein 1 3 1368889_at NM_023101 SNARE Vti1a-beta protein 4 1387046_at NM_053792 selective LIM binding factor 5 1371192_at BF566236 neurofibromatosis 2 6 1369195_at NM_013068 Fatty acid binding protein 2 7 1398540_at BM386789 Rattus norvegicus transcribed sequences 8 1375464_at BI290815 Rattus norvegicus transcribed sequences 9 1387119_at AW433971 mevalonate kinase 10 1374034_at BG379410 Rattus norvegicus transcribed sequence with strong similarity to protein sp: P49589 (H. sapiens) SYC_HUMAN CYSTEINYL-TRNA SYNTHETASE (CYSTEINE--TRNA LIGASE) (CYSRS) 11 1392633_at AI045724 Rattus norvegicus transcribed sequences 12 1371027_at BF556820 Cas-Br-M (murine) ectropic retroviral transforming sequence b 13 1376708_at BM385170 Rattus norvegicus transcribed sequences 14 1387220_at NM_019323 mast cell protease 9 15 1377034_at BF411331 "Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_109591.1 (H. sapiens) serine (or cysteine) proteinase inhibitor, clade B (ovalbumin), member 1; protease inhibitor 2 (anti-elastase), monocyte/neutrophil; protease inhibitor 16 1374324_at AA945828 Rattus norvegicus transcribed sequences 17 1370510_a_at AB012600 aryl hydrocarbon receptor nuclear translocator-like 18 1387130_at NM_133315 "solute carrier family 39 (iron-regulated transporter), member 1" 19 1390562_s_at BE102350 Rattus norvegicus transcribed sequences 20 1390960_at AA893602 Rattus norvegicus transcribed sequences 21 1371925_at AA893621 Rattus norvegicus clone C201 intestinal epithelium proliferating cell-associated mRNA sequence 22 1370693_a_at M18630 cyclic nucleotide phosphodiesterase 1 23 1377036_at BE102350 Rattus norvegicus transcribed sequences 24 1392794_at AA893579 Rattus norvegicus transcribed sequences 25 1388694_at AI233121 MHC class I RT1.O type 149 processed pseudogene 26 1388754_at AI176839 Rattus norvegicus transcribed sequences 27 1368233_at NM_031042 "general transcription factor IIF, polypeptide 2 (30 kD subunit)" 28 1368679_a_at L14782 lyn protein non-receptor kinase 29 1372177_at AI180033 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_004522.1 (H. sapiens) molybdenum cofactor synthesis 2 [Homo sapiens] 30 1372868_at BF284295 Rattus norvegicus transcribed sequence with weak similarity to protein sp: O14657 (H. sapiens) TO1B_HUMAN Torsin B precursor 31 1380472_at AI639486 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_003860.1 (H. sapiens) carboxylesterase 2; intestinal carboxylesterase; liver carboxylesterase-2 [Homo sapiens] 32 1387994_at U89280 oxidative 17 beta hydroxysteroid dehydrogenase type 6 33 1390531_at BE098021 Rattus norvegicus transcribed sequences 34 1388055_at U39207 cytochrome P450 4F5 35 1368379_at NM_054001 "CD36 antigen (collagen type I receptor, thrombospondin receptor)-like 2" 36 1368101_at NM_012518 calmodulin 3 37 1389302_at BI289482 Rattus norvegicus transcribed sequence with weak similarity to protein pir: S30833 (S. cerevisiae) S30833 hypothetical protein YEL044w - yeast (Saccharomyces cerevisiae) 38 1389378_at AI599324 Rattus norvegicus transcribed sequence with weak similarity to protein pir: A42973 (H. sapiens) A42973 serum protein MSE55 - human 39 1387444_at NM_133592 brain-enriched membrane-associated protein tyrosine BEM-2 40 1374284_at AI227769 Rattus norvegicus transcribed sequences 41 1371239_s_at AF053361 "tropomyosin 3, gamma" 42 1387596_at NM_053897 "Proteinase-activated receptor-2, G protein-coupled receptor 11" 43 1389873_at BI282953 apoptosis-associated speck-like protein containing a CARD 44 1369773_at NM_017105 Bone morphogenetic protein 3 45 1374119_at BI279615 "Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_004424.1 (H. sapiens) E74- like factor 3 (ets domain transcription factor, epithelial- specific); E74-like factor 3 (ets domain transcription factor); ets domain transcription 46 1368128_at NM_031598 "phospholipase A2, group IIA (platelets, synovial fluid)" 47 1368270_at NM_012907 Apolipoprotein B editing protein 48 1367960_at NM_019186 ADP-ribos lation-like 4 49 1367942_at NM_019144 acid phosphatase 5 50 1392694_at AW526101 Rattus norvegicus transcribed sequences 51 1374452_at BF399743 phosphodiesterase 9A 52 1368727_at NM_053929 "solute carrier family 7 (cationic amino acid transporter, y+ system), member 9" 53 1376625_at BI296015 Rattus norvegicus transcribed sequences 54 1367768_at NM_031655 latexin 55 1369193_at AF474979 cyclin dependent kinase inhibitor 2B 56 1382714_at AA875186 Rattus norvegicus transcribed sequences 57 1376359_at BG375355 "Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_113645.1 (H. sapiens) membrane-spanning 4-domains, subfamily A, member 8B [Homo sapiens]" 58 1372064_at BI296385 Rattus norvegicus CDK104 mRNA 59 1388396_at BI275932 Rattus norvegicus transcribed sequence with strong similarity to protein sp: O00506 (H. sapiens) ST25_HUMAN Serine/threonine protein kinase 25 (Sterile 20/oxidant stress-response kinase 1) (Ste20/oxidant stress response kinase-1) (SOK-1) (Ste20- like kinase) 60 1384191_at BF387765 Rattus norvegicus transcribed sequences 61 1372255_at BF283284 Rattus norvegicus transcribed sequence with strong similarity to protein sp: P54136 (H. sapiens) SYR_HUMAN ARGINYL-TRNA SYNTHETASE (ARGININE--TRNA LIGASE) (ARGRS) 62 1369262_at NM_022277 caspase-8 63 1376117_at BI289103 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_079533.1 (H. sapiens) NG22 protein; choline transporter-like protein 4 [Homo sapiens] 64 1380262_at AA893436 Rattus norvegicus transcribed sequences 65 1370113_at NM_023987 inhibitor of apoptosis protein 1 66 1368437_at NM_019174 carbonic anhydrase 4 67 1390455_at AI013474 Rattus norvegicus transcribed sequence with strong similarity to protein sp: P08910 (H. sapiens) HPS1 HUMAN Protein PHPS1-2 68 1378658_at BI292185 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_036260.1 (H. sapiens) calcium activated chloride channel 4 [Homo sapiens] 69 1376976_at AI009823 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_002995.1 (H. sapiens) secreted and transmembrane 1 precursor; K12 protein perecursor; type 1a transmembrane protein [Homo sapiens] 70 1370706_a_at U39943 cytochrome P450 monooxygenase 71 1372997_at AI105243 Rattus norvegicus transcribed sequences 72 1369183_at NM_019231 mitogen activated protein kinase 13 73 1386917_at NM_012744 Pyruvate carboxylase 74 1390678_at AA955527 Rattus norvegicus transcribed sequences 75 1398847_at BG376935 diphosphoinositol polyphosphate phosphohydolase type II 76 1368073_at NM_012591 Interferon regulatory factor 1 77 1371394_x_at BG664827 "Rattus norvegicus endogenous retrovirus mRNA, partial sequence" 78 1370422_at AF036537 homocysteine respondent protein HCYP2 79 1375230_at AA800192 "Rattus norvegicus endogenous retrovirus mRNA, partial sequence" 80 1368007_at NM_022849 deleted in malignant brain tumors 1 81 1372671_at BI284293 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_060809.1 (H. sapiens) hypothetical protein FLJ11149 [Homo sapiens] 82 1368219_at NM_017137 chloride channel 2 83 1367760_at D13341 mitogen activated protein kinase kinase 1 84 1388006_at U89744 putative cell surface antigen 85 1367925_at NM_022715 major vault protein 86 1390128_at BF557618 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_065145.1 (H. sapiens) CHMP1.5 protein [Homo sapiens] 87 1388745_at AI228417 "Rattus norvegicus transcribed sequence with strong similarity to protein pdb: 1BGM (E. coli) O Chain O, Beta-Galactosidase (Chains I-P)" 88 1369940_at NM_031811 transaldolase 1 89 1370400_at L23128 "Rattus norvegicus MHC class I mRNA, complete cds" 90 1371970_at AA799328 Rattus norvegicus transcribed sequences 91 1386933_at NM_134418 secretory (zymogen) granule membrane glycoprotein GP2 92 1369100_at NM_134375 angiotensin/vasopressin receptor 93 1372619_at AI172185 "Rattus norvegicus Aa2-277 mRNA, complete cds" 94 1380129_at AA818937 Rattus norvegicus transcribed sequence with weak similarity to protein sp: P10266 (H. sapiens) POL1_HUMAN Endogenous retrovirus HERV-K10 putative pol polyprotein [Includes: Reverse transcriptase; Endonuclease] 95 1371078_at AI500830 RT1 class Ib gene 96 1367586_at NM_017025 lactate dehydrogenase A 97 1374033_at BG373505 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: I38135 (H. sapiens) I38135 multicatalytic endopeptidase complex (EC 3.4.99.46) beta chain MECL-1 - human 98 1373913_at BF282271 Rattus norvegicus transcribed sequence with strong similarity to protein pir: T50626 (H. sapiens) T50626 hypothetical protein DKFZp762K1914.1 - human (fragment) 99 1372665_at AI230228 "Rattus norvegicus phosphoserine aminotransferase mRNA, complete cds" 100 1376056_at BF291214 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_116178.1 (H. sapiens) hypothetical protein FLJ14464 [Homo sapiens] 101 1368066_at NM_053812 BCL2-antagonist/killer 1 102 1374838_at BI293504 Rattus norvegicus transcribed sequence with weak similarity to protein sp: Q13342 (H. sapiens) LY10_HUMAN LYSP100 protein (Lymphoid-restricted homolog of Sp100) (Nuclear autoantigen Sp-140) (Speckled 140 kDa) (Nuclear body protein Sp140) 103 1372816_at BE107319 Rattus norvegicus transcribed sequences 104 1388236_x_at M24026 RT1 class Ib gene 105 1371210_s_at AJ276126 RT1 class Ib gene 106 1390325_at BI289418 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_083693.1 (M. musculus) RIKEN cDNA 9030605E16 [Mus musculus] 107 1369279_at NM_130819 retinol dehydrogenase homolog 108 1390021_at BM391206 histone 2b 109 1368317_at NM_019157 aquaporin 7 110 1387100_at NM_031703 aquaporin 3 111 1368975_at NM_013127 CD38 antigen 112 1386908_at NM_022278 glutaredoxin 1 (thioltransferase) 113 1369427_at NM_022617 macrophage expressed gene 1 114 1369110_x_at NM_012645 RT1 class Ib gene 115 1369957_at NM_019341 regulator of G-protein signaling 5 116 1398985_at AI716480 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_004277.1 (H. sapiens) GTP binding protein 1 [Homo sapiens] 117 1368413_at NM_022935 Amiloride binding protein 1 118 1370960_at BE104060 insulin-like growth factor-binding protein 5 119 1368505_at NM_017214 regulator of G-protein signaling 4 120 1373386_at AI179953 Rattus norvegicus transcribed sequences 121 1387348_at BE113270 insulin-like growth factor-binding protein 5 122 1370638_at AF069525 ankyrin 3 (G) 123 1377112_at AA859352 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_001776.1 (H. sapiens) cytidine deaminase [Homo sapiens] 124 1389611_at AA849857 Very low density lipoprotein receptor 125 1369098_at NM_013155 Very low density lipoprotein receptor 126 1368294_at NM_053907 deoxyribonuclease I-like 3 127 1368342_at NM_031544 Adenosine monophosphate deaminase 3 128 1368965_at NM_030834 monocarboxylate transporter 129 1392819_at AW921478 "Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_071744.1 (H. sapiens) CD20- like precusor; membrane-spanning 4-domains, subfamily A, member 6 [Homo sapiens]" 130 1367774_at NM_031509 "glutathione S-transferase, alpha 1" 131 1387687_at NM_133542 "immunoglobulin superfamily, member 6" 132 1369173_at NM_032060 complement component 3a receptor 1 133 1372013_at BG380285 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_006426.1 (H. sapiens) interferon induced transmembrane protein 2 (1-8D);
interferon-inducible [Homo sapiens] 134 1373025_at AI411618 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: C1HUQC (H. sapiens) C1HUQC complement subcomponent C1q chain C precursor - human 135 1376652_at BF418957 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: C1HUQA (H. sapiens) C1HUQA complement subcomponent C1q chain A precursor - human 136 1368420_at NM_012532 ceruloplasmin 137 1373932_at BE098739 Rattus norvegicus transcribed sequences 138 1387893_at D88250 "complement component 1, s subcomponent" 139 1388557_at BF284922 Rattus norvegicus transcribed sequences 140 1374730_at AI102519 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_003323.1 (H. sapiens) TYRO protein tyrosine kinase binding protein; polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy [Homo sapiens] 141 1368000_at NM_016994 Complement component 3 142 1368558_s_at NM_017196 allograft inflammatory factor 1 143 1373523_at AI011757 Rattus norvegicus transcribed sequence with weak similarity to protein sp: P08637 (H. sapiens) FC3A_HUMAN Low affinity immunoglobulin gamma FC region receptor III-A precursor (IGG FC receptor III- 2) (FC-gamma RIII-alpha) (FC-gamma RIIIA) (FCRIIIA) (FC-gamm 144 1387011_at NM_130741 lipocalin 2 145 1370885_at AA849399 cathepsin Y 146 1389470_at AI639117 Complement component 2 147 1368430_at AF154349 "protease, cysteine, 1 (legumain)" 148 1375010_at AI177761 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_001242.1 (H. sapiens) CD68 antigen; Macrophage antigen CD68 (microsialin) [Homo sapiens] 149 1389006_at AI170394 Rattus norvegicus transcribed sequences 150 1373575_at BE111722 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: A35241 (H. sapiens) A35241 IgE Fc receptor gamma chain precursor - human 151 1390510_at BI294706 "Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_071744.1 (H. sapiens) CD20- like precusor; membrane-spanning 4-domains, subfamily A, member 6 [Homo sapiens]" 152 1376390_at BF395317 "Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_071744.1 (H. sapiens) CD20- like precusor; membrane-spanning 4-domains, subfamily A, member 6 [Homo sapiens]" 153 1368187_at NM_133298 glycoprotein (transmembrane) nmb 154 1389553_at BF393825 "Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_057268.1 (H. sapiens) C-type (calcium dependent, carbohydrate-recognition domain) lectin, superfamily member 6; dendritic cell immunoreceptor; C-type lectin [Homo sapiens]" 155 1367568_a_at NM_012862 matrix Gla protein 156 1370628_at M34097 granzyme B 157 1379604_at BF284937 "Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_085146.1 (H. sapiens) apolipoprotein L, 4 [Homo sapiens]" 158 1368464_at NM_022393 macrophage galactose N-acetyl-galactosamine specific lectin 159 1379766_at AI500952 Rattus norvegicus transcribed sequences 160 1370516_at AB026665 peptide/histidine transporter PHT2 161 1373071_at AI103101 Rattus norvegicus transcribed sequence with strong similarity to protein pir: T00702 (H. sapiens) T00702 hypothetical protein F25965 1 - human (fragment) 162 1393224_at AW529774 Rattus norvegicus transcribed sequences 163 1390312_at BG670441 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_060124.1 (H. sapiens) hypothetical protein FLJ20073 [Homo sapiens] 164 1377412_at AI146237 Rattus norvegicus transcribed sequences 165 1399125_at BI275516 Rattus norvegicus transcribed sequence with weak similarity to protein sp: P49441 (H. sapiens) INPP_HUMAN Inositol polyphosphate 1-phosphatase (IPPase) (IPP) 166 1386986_at NM_053340 opioid growth factor receptor 167 1374731_at BI275929 "Rattus norvegicus transcribed sequence with moderate similarity to protein pdb: 1LBG (E. coli) B Chain B, Lactose Operon Repressor Bound To 21-Base Pair Symmetric Operator Dna, Alpha Carbons Only" 168 1388880_at BI278962 Lysosomal associated membrane protein 1 (120 kDa) 169 1376075_at BM385544 Rattus norvegicus transcribed sequences 170 1369559_a_at NM_019195 integrin-associated protein 171 1376972_at AI407028 "solute carrier family 39 (iron-regulated transporter), member 1" 172 1388776_at AI169176 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_057563.1 (H. sapiens) hypothetical protein LOC51246 [Homo sapiens] 173 1376029_at BI295991 Rattus norvegicus transcribed sequences 174 1388164_at AF029241 "Rat MHC class I RT1.C/E mRNA, 3' end" 175 1389734_x_at BI282965 RT1 class Ib gene 176 1370429_at L40362 RT1 class Ib gene 177 1388900_at BG381414 Rattus norvegicus transcribed sequences 178 1375955_at BI289415 Rattus norvegicus transcribed sequences 179 1389011_at BG374333 Rattus norvegicus transcribed sequences 180 1389387_at BF561377 hydroxyindole-O-methyltransferase 181 1387206_at NM_031740 "UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase, polypeptide 6" 182 1388071_x_at M24024 RT1 class Ib gene 183 1398925_at BI274664 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_060818.1 (H. sapiens) hypothetical protein FLJ11171 [Homo sapiens] 184 1368224_at NM_031531 Serine protease inhibitor 185 1374718_at AA945915 Rattus norvegicus transcribed sequences 186 1394340_at BF523172 Rattus norvegicus transcribed sequence with weak similarity to protein sp: P49441 (H. sapiens) INPP_HUMAN Inositol polyphosphate 1-phosphatase (IPPase) (IPP) 187 1376022_at BI292196 Rattus norvegicus transcribed sequences 188 1368732_at NM_032056 "transporter 2, ATP-binding cassette, sub-family B (MDR/TAP)" 189 1367710_at NM_017257 "protease (prosome, macropain) 28 subunit, beta" 190 1367786_at NM_080767 "proteasome (prosome, macropain) subunit, beta type, 8 (low molecular mass polypeptide 7)" 191 1389170_at BF283754 Rattus norvegicus transcribed sequences 192 1375853_at BF284358 Rattus norvegicus transcribed sequences 193 1373757_at AW529298 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_006691.1 (H. sapiens) FLN29 gene product [Homo sapiens] 194 1388149_at X57523 "transporter 1, ATP-binding cassette, sub-family B (MDR/TAP)" 195 1372604_at BI289459 "Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_055164.1 (H. sapiens) apolipoprotein L, 3; TNF-inducible protein CG12-1 [Homo sapiens]" 196 1387027_a_at U72741 "Lectin, galactose binding, soluble 9 (Galectin-9)" 197 1372034_at BF284106 Rattus norvegicus transcribed sequences 198 1370186_at AI599350 "proteosome (prosome, macropain) subunit, beta type 9 (large multifunctional protease 2)" 199 1388212_a_at AJ243974 "Rat MHC class I RT1.C/E mRNA, 3' end" 200 1388213_a_at AJ243973 "Rat MHC class I RT1.C/E mRNA, 3' end" 201 1371123_x_at AJ243973 "Rat MHC class I RT1.C/E mRNA, 3' end" 202 1371152_a_at Z18877 25 oligoadenylate synthetase 203 1372930_at AI411381 "Rattus norvegicus cDNA, clone: aC10, differentially expressed in pylorus" 204 1388791_at BI275911 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: T14738 (H. sapiens) T14738 hypothetical protein DKFZp564A2416.1 - human (fragment) 205 1369716_s_at NM_012976 "Lectin, galactose binding, soluble 5 (Galectin-5)" 206 1376496_at AI717736 "Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_112092.1 (H. sapiens) apolipoprotein L, 2 [Homo sapiens]" 207 1374337_at AI408954 "Rattus norvegicus transcribed sequence with moderate similarity to protein pdb: 1LBG (E. coli) B Chain B, Lactose Operon Repressor Bound To 21-Base Pair Symmetric Operator Dna, Alpha Carbons Only" 208 1387242_at NM_019335 "Protein kinase, interferon-inducible double stranded RNA dependent" 209 1376144_at AA819679 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_113646.1 (H. sapiens) B aggressive lymphoma gene [Homo sapiens] 210 1368835_at AW434718 signal transducer and activator of transcription 1 211 1387946_at AF065438 peptidylprolyl isomerase C-associated protein 212 1383564_at BF411036 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_004022.1 (H. sapiens) interferon regulatory factor 7 isoform d [Homo sapiens] 213 1389034_at BI295179 Rattus norvegicus transcribed sequence with weak similarity to protein sp: Q9UMW8 (H. sapiens) UBPI_HUMAN Ubiquitin carboxyl-terminal hydrolase 18 (Ubiquitin thiolesterase 18) (Ubiquitin-specific processing protease 18) (Deubiquitinating enzyme 18) (Ubiqui 214 1388347_at AI233210 Rattus norvegicus transcribed sequence with weak similarity to protein sp: Q16553 (H. sapiens) LY6E_HUMAN Lymphocyte antigen Ly-6E precursor (Retinoic acid-induced gene E protein) (RIG-E) (Thymic shared antigen-1) (TSA-1) (Stem cell antigen 2) (SCA-2) 215 1376845_at AA819034 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_114425.1 (H. sapiens) TLH29 protein precursor [Homo sapiens] 216 1376693_at AA998964 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_060124.1 (H. sapiens) hypothetical protein FLJ20073 [Homo sapiens] 217 1387770_at NM_130743 "interferon, alpha-inducible protein 27-like" 218 1376920_at BF408536 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_060124.1 (H. sapiens) hypothetical protein FLJ20073 [Homo sapiens] 219 1387354_at NM_032612 signal transducer and activator of transcription 1 220 1373037_at BI279216 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_004214.1 (H. sapiens) ubiquitin-conjugating enzyme E2L 6 [Homo sapiens] 221 1387995_a_at BI285494 interferon induced transmembrane protein 3-like 222 1377497_at BF419319 "Rattus norvegicus transcribed sequence with moderate similarity to protein pir: I60307 (E. coli) I60307 beta- galactosidase, alpha peptide - Escherichia coli" 223 1368227_at NM_031664 "solute carrier family 28, member 2" 224 1390507_at BI296097 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_002192.2 (H. sapiens) interferon stimulated gene (20 kD) [Homo sapiens] 225 1371440_at AW916647 Prostaglandin F receptor 226 1369590_a_at NM_024134 DNA-damage inducible transcript 3 227 1369202_at NM_017028 myxovirus (influenza virus) resistance 2 228 1374627_at BF283727 Rattus norvegicus transcribed sequences 229 1374551_at BM388891 Rattus norvegicus transcribed sequence with weak similarity to protein pir: JC5262 (H. sapiens) JC5262 leucine zipper protein IFP35 - human 230 1387134_at NM_053687 schlafen 4 231 1373514_at AA899109 Rattus norvegicus transcribed sequences 232 1372585_at BM388445 Rattus norvegicus transcribed sequences 233 1369031_at NM_053374 interferon gamma inducing factor binding protein 234 1371070_at AJ302054 tumor stroma and activated macrophage protein DLM-1 235 1367595_s_at NM_012512 Beta-2-microglobulin 236 1367663_at NM_017264 "protease (prosome, macropain) 28 subunit, alpha" 237 1390042_at AI071166 Rattus norvegicus transcribed sequences 238 1369186_at D85899 caspase 1 239 1389571_at BG666368 Rattus norvegicus transcribed sequence with moderate similarity to protein sp: P52630 (H. sapiens) STA2_HUMAN Signal transducer and activator of transcription 2 (p113) 240 1370913_at AI409634 Best5 protein 241 1387969_at U22520 chemokine (C-X-C motif) ligand 10 242 1376908_at AW531805 Rattus norvegicus transcribed sequence with weak similarity to protein sp: O14879 (H. sapiens) IFT4_HUMAN Interferon-induced protein with tetratricopeptide repeats 4 (IFIT-4) (Interferon-induced 60 kDa protein) (IFI-60K) (ISG-60) (CIG49) (Retinoic acid-ind 243 1369973_at NM_017154 xanthine dehydrogenase 244 1368332_at NM_133624 "guanylate binding protein 2, interferon-inducible" 245 1372254_at AW915763 Rattus norvegicus transcribed sequence with moderate similarity to protein sp: P05155 (H. sapiens) IC1_HUMAN
Plasma protease C1 inhibitor precursor (C1 Inh) (C1Inh) 246 1373197_at AI578263 Rattus norvegicus transcribed sequences 247 1370892_at BI285347 complement component 4a 248 1372757_at BM386875 signal transducer and activator of transcription 1 249 1376151_a_at AI407953 Rattus norvegicus transcribed sequences 250 1387283_at NM_134350 myxovirus (influenza virus) resistance 2 251 1371015_at X52711 myxovirus (influenza virus) resistance 252 1373992_at AI408440 "Rattus norvegicus cDNA, clone: aD9, differentially expressed in pylorus" 253 1390738_at BM385476 Rattus norvegicus mRNA for DAMP-1 protein 254 1388056_at AF068268 2'5' oligoadenylate synthetase-2 255 1389365_at AI228291 Rattus norvegicus transcribed sequences 256 1369726_at NM_033098 TAP binding protein 257 1370172_at AA892254 superoxide dismutase 2 258 1386893_at NM_017113 granulin 259 1375796_at BI300770 "Rattus norvegicus Ac2-233 mRNA, complete cds" 260 1389014_at BI297612 pre-B-cell colony-enhancing factor 261 1371209_at AJ243338 RT1 class Ib gene 262 1388255_x_at AJ243338 RT1 class Ib gene 263 1373406_at BM384991 Rattus norvegicus transcribed sequences 264 1375006_at BE121050 Rattus norvegicus transcribed sequences 265 1387897_at L16532 cyclic nucleotide phosphodiesterase 1 266 1369456_at NM_017250 5-hydroxytryptamine (serotonin) receptor 2B 267 1374141_at BG372419 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_071387.1 (H. sapiens) chromosome 20 open reading frame 67 [Homo sapiens] 268 1374678_at BE109578 Rattus norvegicus transcribed sequences 269 1372968_at BM385950 Rattus norvegicus transcribed sequence with weak similarity to protein pir: S37032 (R. norvegicus) S37032 gene LL5 protein - rat 270 1371832_at AW526333 Rattus norvegicus transcribed sequences 271 1390117_at BG372455 Rattus norvegicus transcribed sequences 272 1369381_a_at D50306 "solute carrier family 15 (oligopeptide transporter), member 1" 273 1388103_at AF361355 voltage-dependent calcium channel gamma subunit-like protein 274 1386943_at NM_022533 plasmolipin 275 1373060_at AI406281 Rattus norvegicus transcribed sequence with strong similarity to protein pir: T12468 (H. sapiens) T12468 hypothetical protein DKFZp564O123.1 - human 276 1375911_at AI171772 "Rattus norvegicus hypothetical protein LK44 mRNA, complete cds" 277 1370071_at NM_130399 Adenosine deaminase 278 1368762_at NM_053299 ubiquitin D 279 1369712_at NM_031735 serine/threonine kinase 3 280 1368419_at AF202115 ceruloplasmin 281 1369029_at NM_057194 phospholipid scramblase 1 282 1369888_at NM_012707 glucagon 283 1398879_at BE329031 arrestin-E 284 1369942_at NM_031675 actinin alpha 4 285 1370807_at AF411216 vacuole Membrane Protein 1 286 1377124_at AA964824 Rattus norvegicus transcribed sequence with weak similarity to protein pir: S48059 (H. sapiens) S48059 metal-regulatory transcription factor - human 287 1390221_at BM385216 Rattus norvegicus transcribed sequences 288 1390604_s_at BM387863 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_055103.1 (H. sapiens) integrin beta 3 binding protein (beta3-endonexin); beta 3 endonexin [Homo sapiens] 289 1398916_at AI104388 heat shock 27 kDa protein 1 290 1374600_at BM986536 germinal histone H4 gene 291 1370307_at M64780 Agrin 292 1371888_at AA892843 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_078816.1 (H. sapiens) hypothetical protein FLJ20917 [Homo sapiens] 293 1373603_at BG673166 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: T45061 (H. sapiens) T45061 hypothetical protein c316G12.2 [imported]- human 294 1388408_at AA800199 Rattus norvegicus transcribed sequence with weak similarity to protein pir: T34520 (H. sapiens) T34520 hypothetical protein DKFZp564J157.1 - human (fragment) 295 1368471_at NM_013118 guanylate cyclase activator 2A 296 1374401_at AI549249 Rattus norvegicus transcribed sequence with strong similarity to protein sp: O60749 (H. sapiens) SNX2_HUMAN Sorting nexin 2 297 1371267_at M64795 zzN/A 298 1370832_at U06434 small inducible cytokine A4 299 1383241_at BI292425 Rattus norvegicus transcribed sequence with moderate similarity to protein sp: P00736 (H. sapiens) C1R_HUMAN Complement C1r component precursor 300 1387922_at AF109674 late gestation lung protein 1 301 1376447_at AI706850 Rattus norvegicus transcribed sequence with weak similarity to protein sp: P10163 (H. sapiens) PRB4_HUMAN Salivary proline-rich protein PO precursor (Allele S) 302 1367740_at M14400 "creatine kinase, brain" 303 1387036_at NM_024360 hairy and enhancer of split 1 (Drosophila) 304 1370360_at AF146738 testis specific protein 305 1389111_at BF396316 Rattus norvegicus transcribed sequences 306 1374454_at BM388557 Rattus norvegicus transcribed sequences 307 1367764_at NM_012923 Cyclin G1 308 1368038_at AF260258 synaptojanin 2 binding protein 309 1372635_at BF282163 Rattus norvegicus transcribed sequences 310 1373571_at AI170276 Rattus norvegicus transcribed sequences 311 1375872_at AI144944 Rattus norvegicus transcribed sequences 312 1383767_at AW524430 Rattus norvegicus transcribed sequences 313 1376325_at BM388975 Rattus norvegicus transcribed sequence with strong similarity to protein pir: S52863 (H. sapiens) S52863 DNA-binding protein R kappa B - human 314 1388485_at BG380414 Rattus norvegicus transcribed sequences 315 1368782_at NM_019348 somatostatin receptor 2 316 1375867_at AW524493 Rattus norvegicus transcribed sequences 317 1375138_at AA893169 Tissue inhibitor of metalloproteinase 3 318 1390112_at BF284634 "Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_004096.2 (H. sapiens) EGF- containing fibulin-like extracellular matrix protein 1 precursor, isoform a precursor; fibrillin-like [Homo sapiens]" 319 1372935_at AI598550 Rattus norvegicus transcribed sequences 320 1368322_at NM_012880 superoxide dismutase 3 321 1372104_at BF289002 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_003106.1 (H. sapiens) UDP- N-acteylglucosamine pyrophosphorylase 1; AgX; sperm associated antigen 2; UDP-N-acteylglucosamine pyrophosphorylase 1; Sperm associated antigen 2 [Hom 322 1367687_a_at M25719 Peptidylglycine alpha-amidating monooxygenase 323 1372455_at AI410264 Rattus norvegicus transcribed sequence with strong similarity to protein sp: O95859 (H. sapiens) TNE2_HUMAN Tetraspan NET-2 324 1372940_at BM389329 Rattus norvegicus transcribed sequences 325 1367631_at NM_022266 connective tissue growth factor 326 1376344_at AI010267 Rattus norvegicus transcribed sequences 327 1370408_at AF313411 putative small membrane protein NID67 328 1390283_at BI274636 Rattus norvegicus transcribed sequences 329 1372327_at BF552877 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_057216.1 (H. sapiens) myelin gene expression factor 2 [Homo sapiens] 330 1376175_at BF283433 Rattus norvegicus transcribed sequences 331 1373966_at BF406242 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: T43490 (H. sapiens) T43490 hypothetical protein DKFZp434A139.1 - human (fragments) 332 1376749_at AA945955 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: B35272 (H. sapiens) B35272 osteoinductive factor - human 333 1367604_at NM_022501 cysteine-rich protein 2 334 1398345_at BM389225 angiopoietin-like 2 335 1389836_a_at AI599265 Tissue inhibitor of metalloproteinase 3 336 1389256_at BG381256 Rattus norvegicus transcribed sequences 337 1371500_at BG375362 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_003564.1 (H. sapiens) latent transforming growth factor beta binding protein 4 [Homo sapiens] 338 1371629_at AI105444 "Rattus norvegicus transcribed sequence with moderate similarity to protein pir: I60307 (E. coli) I60307 beta- galactosidase, alpha peptide - Escherichia coli" 339 1387707_at NM_017102 "solute carrier family 2, member 2" 340 1387505_at NM_013145 "Guanine nucleotide binding protein, alpha inhibiting 1" 341 1388890_at BM390316 Rattus norvegicus transcribed sequences 342 1373506_at AA944568 Rattus norvegicus transcribed sequences 343 1369415_at NM_053328 "basic helix-loop-helix domain containing, class B2" 344 1372264_at BI277460 "Rattus norvegicus transcribed sequence with strong similarity to protein sp: P35558 (H. sapiens) PPCC_HUMAN Phosphoenolpyruvate carboxykinase, cytosolic [GTP] (Phosphoenolpyruvate carboxylase) (PEPCK-C)" 345 1374105_at H31665 Rattus norvegicus transcribed sequences 346 1387156_at NM_024391 17-beta hydroxysteroid dehydrogenase type 2 347 1376569_at BM385790 Rattus norvegicus transcribed sequence with strong similarity to protein sp: Q9Y5W3 (H. sapiens) KLF2_HUMAN Kruppel-like factor 2 (Lung kruppel-like factor) 348 1387028_a_at M86708 "Inhibitor of DNA binding 1, helix-loop-helix protein (splice variation)" 349 1376661_at AI556122 Rattus norvegicus transcribed sequences 350 1377287_at AA957673 Rattus norvegicus transcribed sequences 351 1375910_at AA874943 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: T46465 (H. sapiens) T46465 hypothetical protein DKFZp434A0530.1 - human 352 1388447_at AA800701 Rattus norvegicus transcribed sequences 353 1370912_at BI278231 R. norvegicus hsp70.2 mRNA for heat shock protein 70 354 1368883_at NM_030868 NOV protein 355 1388945_at BM385779 "Rattus norvegicus transcribed sequence with moderate similarity to protein pir: I60307 (E. coli) I60307 beta- galactosidase, alpha peptide - Escherichia coli" 356 1389514_at AI711152 Rattus norvegicus transcribed sequences 357 1377086_at AI233530 Rattus norvegicus transcribed sequences 358 1373947_at BI278545 Rattus norvegicus transcribed sequence with strong similarity to protein pir: A47220 (H. sapiens) A47220 dermatopontin precursor - human 359 1374171_at AI170507 "ATP-binding cassette, sub-family C (CFTR/MRP), member 9" 360 1372490_at BF283759 Rattus norvegicus transcribed sequences 361 1390257_at BG376956 Rattus norvegicus transcribed sequences 362 1369651_at NM_012673 Thymus cell surface antigen 363 1388866_at AA799392 "Rattus norvegicus transcribed sequence with moderate similarity to protein pdb: 1LBG (E. coli) B Chain B, Lactose Operon Repressor Bound To 21-Base Pair Symmetric Operator Dna, Alpha Carbons Only" 364 1368303_at NM_031678 period homolog 2 365 1370399_at M29853 "cytochrome P450, subfamily 4B, polypeptide 1" 366 1385606_at BF559626 Rattus norvegicus transcribed sequences 367 1376170_at BI290821 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: S46657 (H. sapiens) S46657 collagen alpha 1(XIV) chain - human (fragments) 368 1376832_at AA800763 Rattus norvegicus transcribed sequences 369 1376049_at AA925805 Rattus norvegicus transcribed sequences 370 1398365_at AI171466 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_057048.1 (H. sapiens) CGI- 38 protein [Homo sapiens] 371 1372847_at AW524458 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_064571.1 (H. sapiens) DC11 protein [Homo sapiens] 372 1372872_at BI291271 Rattus norvegicus transcribed sequences 373 1376177_at AI179609 Rattus norvegicus transcribed sequences 374 1387039_at NM_030828 glypican 1 375 1372208_at AA942959 "protein phosphatase 1, regulatory (inhibitor) subunit 1B" 376 1376933_at AI170377 Rattus norvegicus transcribed sequences 377 1388034_at AB070355 kinesin family member 1B 378 1371794_at BM391449 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: T14273 (M. musculus) T14273 zinc finger protein 106 - mouse 379 1393464_at BM383378 Rattus norvegicus transcribed sequences 380 1389007_at AI231799 Rattus norvegicus transcribed sequences
381 1371508_at AI412098 "protein tyrosine phosphatase type IVA, member 2" 382 1374694_at BF284602 Rattus norvegicus transcribed sequences 383 1374558_at AI010316 Rattus norvegicus transcribed sequences 384 1399028_at AI171954 Rattus norvegicus transcribed sequences 385 1368550_at NM_022858 HNF-3/forkhead homolog-1 386 1370211_at BE106940 neurogranin 387 1387090_a_at NM_024135 LIM motif-containing protein kinase 2 388 1370072_at NM_012608 membrane metallo endopeptidase 389 1371703_at AI407114 Complement component 3 390 1390319_at BG378301 Rattus norvegicus transcribed sequence 391 1387008_at NM_022948 tricarboxylate carrier-like protein 392 1389561_at BE110624 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_079740.1 (M. musculus) RIKEN cDNA 1810021J13 [Mus musculus] 393 1376023_at BM386555 Rattus norvegicus transcribed sequences 394 1386979_at NM_133395 developmentally regulated protein TPO1 395 1368163_at J02997 Dipeptidyl peptidase 4 396 1374935_at AI412099 "Rattus norvegicus transcribed sequence with moderate similarity to protein pir: I60307 (E. coli) I60307 beta- galactosidase, alpha peptide - Escherichia coli" 397 1387084_at NM_012789 Dipeptidyl peptidase 4 398 1369770_at NM_012719 somatostatin receptor 1 399 1389601_at BI293610 "Rattus norvegicus transcribed sequence with moderate similarity to protein pir: I60307 (E. coli) I60307 beta- galactosidase, alpha peptide - Escherichia coli" 400 1368295_at NM_080786 "solute carrier family 21 (organic anion transporter), member 9" 401 1376242_at BF402365 Rattus norvegicus transcribed sequences 402 1387059_at NM_019362 "serine threonine kinase 39 (STE20/SPS1 homolog, yeast)" 403 1389310_at BF400694 Rattus norvegicus transcribed sequences 404 1387169_at NM_053400 "transducin-like enhancer of split 3, homolog of Drosophila" 405 1387023_at NM_031154 "glutathione S-transferase, mu type 3 (Yb3)" 406 1398430_at AW524711 Rattus norvegicus transcribed sequences 407 1369249_at NM_053714 progressive ankylosis 408 1387913_at U48220 cytochrome P450 2D18 409 1370320_at AY083160 MAWD binding protein 410 1376047_at BI285321 Rattus norvegicus transcribed sequences 411 1387315_at NM_012878 "surfactant, pulmonary-associated protein D" 412 1387239_a_at AB008803 "peptidyl arginine deiminase, type 4" 413 1371477_at BG380735 Rattus norvegicus transcribed sequences 414 1386974_at NM_134397 LL5 protein 415 1370930_at BF417285 kinesin 1C 416 1371689_at BE107334 eukaryotic translation elongation factor 1 alpha 1 417 1389040_at AI170825 Rattus norvegicus transcribed sequences 418 1390710_x_at AA850618 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_003096.1 (H. sapiens) sortilin-related receptor containing LDLR class A repeats preproprotein; sorting protein-related receptor containing LDLR class A repeats; low-density li 419 1367700_at NM_080698 fibromodulin 420 1388317_at BE110655 Rattus norvegicus transcribed sequences 421 1387184_at NM_024355 axin 2 422 1390399_at BE102391 Rattus norvegicus transcribed sequences 423 1375590_at AA894335 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: T00368 (H. sapiens) T00368 hypothetical protein KIAA0663 - human 424 1387901_at L19180 "protein tyrosine phosphatase, receptor type, D" 425 1388768_at BI285601 Rattus norvegicus transcribed sequences 426 1373434_at AA944179 Rattus norvegicus transcribed sequences 427 1376784_at BI274481 Rattus norvegicus transcribed sequences 428 1390311_at AW528602 Rattus norvegicus transcribed sequences 429 1389150_at AW524559 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_076933.1 (H. sapiens) hypothetical protein MGC3265 [Homo sapiens] 430 1372728_at BE103745 Rattus norvegicus transcribed sequence with moderate similarity to protein sp: Q99523 (H. sapiens) SORT_HUMAN Sortilin precursor (Glycoprotein 95) (Gp95) (Neurotensin receptor 3) (NT3) (100 kDa NT receptor) 431 1377325_a_at AW531278 Rattus norvegicus transcribed sequences 432 1389288_at BI279838 "Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_002479.1 (H. sapiens) NADH dehydrogenase (ubiquinone) 1 alpha subcomplex, 2 (8 kD, B8) [Homo sapiens]" 433 1375951_at AA818521 thrombomodulin 434 1367880_at NM_012974 Laminin chain beta 2 435 1399109_at BI281673 Rattus norvegicus transcribed sequences 436 1369926_at NM_022525 glutathione peroxidase 3 437 1388145_at BM390128 Tenascin X 438 1398973_at BI296499 Rattus norvegicus transcribed sequence with strong similarity to protein sp: Q9HD45 (H. sapiens) T9S3_HUMAN Transmembrane 9 superfamily protein member 3 precursor (SM-11044 binding protein) (EP70- P-iso) 439 1398990_at BI281754 Rattus norvegicus transcribed sequences 440 1371245_a_at BI287300 hemoglobin beta chain complex 441 1392890_at BG663460 Rattus norvegicus transcribed sequences 442 1371102_x_at X05080 hemoglobin beta chain complex 443 1370240_x_at AI179404 "hemoglobin, alpha 1" 444 1367553_x_at NM_033234 hemoglobin beta chain complex 445 1370239_at AI179404 "hemoglobin, alpha 1" 446 1388608_x_at AI577319 "hemoglobin, alpha 1" 447 1388848_at AA891760 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_071934.1 (H. sapiens) hypothetical protein FLJ22056 [Homo sapiens] 448 1389186_at AA944175 "Rattus norvegicus transcribed sequence with moderate similarity to protein pir: I60307 (E. coli) I60307 beta- galactosidase, alpha peptide - Escherichia coli" 449 1371972_at BM388888 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: T00371 (H. sapiens) T00371 hypothetical protein KIAA0668 - human (fragment) 450 1372640_at BI277758 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_076223.1 (M. musculus) RIKEN cDNA 1200009H11 [Mus musculus] 451 1373162_at AI600085 "Rattus norvegicus NYGGF3 mRNA, partial cds" 452 1387310_at NM_134462 putative secretory pathway Ca-ATPase SPCA2 453 1388698_at AI407838 extracellular matrix protein 1 454 1376105_at AI599143 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: S46657 (H. sapiens) S46657 collagen alpha 1(XIV) chain - human (fragments) 455 1373674_at BI283094 Rattus norvegicus transcribed sequence with moderate similarity to protein sp: Q13361 (H. sapiens) MGP2_HUMAN Microfibril-associated glycoprotein 2 precursor (MAGP-2) (MP25) 456 1373952_at AI409841 Rattus norvegicus transcribed sequences 457 1387625_at NM_013104 zzN/A 458 1374674_at AW528792 Rattus norvegicus transcribed sequences 459 1375844_at AI406370 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_067013.1 (H. sapiens) polypyrimidine tract binding protein 2; neural polypyrimidine tract binding protein; PTB-like protein [Homo sapiens] 460 1368407_at NM_022605 heparanase 461 1377076_at AI716131 Rattus norvegicus transcribed sequences 462 1375412_at AI101331 Rattus norvegicus transcribed sequences 463 1376398_at BF417784 Rattus norvegicus transcribed sequences 464 1399054_at BG375798 Rattus norvegicus transcribed sequences 465 1374126_at BG374261 Rattus norvegicus transcribed sequences 466 1368099_at NM_053722 CLIP-associating protein 2 467 1387182_at NM_057201 G protein-coupled receptor 37 (endothelin receptor type B-like) 468 1372814_at BF283084 Rattus norvegicus transcribed sequences 469 1376115_at AA964244 Rattus norvegicus transcribed sequences 470 1369373_at NM_053429 fibroblast growth factor receptor 3 471 1376781_at BI286116 Rattus norvegicus transcribed sequences 472 1369638_at NM_012947 Eukaryotic elongation factor 2 kinase 473 1389003_at BI282008 Rattus norvegicus transcribed sequences 474 1376089_at BI294974 Low density lipoprotein receptor 475 1367932_at NM_017268 3-hydroxy-3-methylglutaryl-Coenzyme A synthase 1 476 1387809_at NM_053703 mitogen-activated protein kinase kinase 6 477 1375549_at AI407719 Rattus norvegicus transcribed sequences 478 1370209_at BE101336 Kruppel-like factor 9 479 1389311_at AW915161 Rattus norvegicus transcribed sequences 480 1390164_at BE099850 Rattus norvegicus transcribed sequences 481 1376989_at BE099568 Rattus norvegicus transcribed sequences 482 1368656_at NM_053856 secretogranin III 483 1373415_at AI407050 Rattus norvegicus transcribed sequences 484 1374819_at AI599945 Rattus norvegicus transcribed sequences 485 1367594_at NM_017087 biglycan 486 1372624_at BF551377 Rattus norvegicus transcribed sequences 487 1376425_at BF420705 "transforming growth factor, beta 2" 488 1389138_at AA945574 Rattus norvegicus transcribed sequences 489 1367940_at NM_053352 chemokine orphan receptor 1 490 1371747_at AI406304 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_077275.1 (H. sapiens) chromosome 20 open reading frame 149 [Homo sapiens] 491 1372978_at BI291218 Rattus norvegicus transcribed sequences 492 1387812_at NM_012999 Subtilisin - like endoprotease 493 1389794_at AI044984 Rattus norvegicus transcribed sequences 494 1388384_at AI407618 Rattus norvegicus transcribed sequences 495 1374557_at BF394235 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_079187.1 (H. sapiens) hypothetical protein FLJ23091 [Homo sapiens] 496 1370579_at U53513 "glycine-, glutamate-, thienylcyclohexylpiperidine- binding protein" 497 1367918_at NM_031066 protein kinase C-binding protein Zeta1 498 1368930_at NM_023021 intermediate conductance calcium-activated potassium channel 499 1387654_at NM_023092 unconventional myosin Myr2 I heavy chain 500 1367759_at NM_012578 Histone H1-0 501 1386957_at NM_053622 nuclear pore membrane glycoprotein 121 kD 502 1388709_at BF284695 Rattus norvegicus transcribed sequence with weak similarity to protein pir: A35804 (H. sapiens) A35804 nucleolin - human 503 1384371_at AW921429 zzN/A 504 1371940_at AW920000 Rattus norvegicus transcribed sequence with strong similarity to protein sp: Q9UPN3 (H. sapiens) ACF7_HUMAN Actin cross-linking family protein 7 (Macrophin) (Trabeculin-alpha) (620 kDa actin-binding protein) (ABP620) 505 1370131_at NM_031556 caveolin 506 1372825_at BI290551 Rattus norvegicus transcribed sequences 507 1367989_at NM_012751 "solute carrier family 2, member 4" 508 1376724_at AI170671 Rattus norvegicus transcribed sequences 509 1372967_at BI280323 Rattus norvegicus transcribed sequences 510 1370291_at AF002281 actinin alpha 2 associated LIM protein 511 1377281_at BM388545 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_065099.1 (H. sapiens) retinitis pigmentosa GTPase regulator interacting protein 1; RPGR-interacting protein [Homo sapiens] 512 1367930_at NM_017195 growth associated protein 43 513 1367628_at NM_019904 "lectin, galactose binding, soluble 1" 514 1388112_at BG666999 solute carrier family 25 (mitochondrial adenine nucleotide translocator) member 4 515 1387873_at BI279661 wap four-disulfide core domain 1 516 1389194_at AI406491 Rattus norvegicus transcribed sequences 517 1388928_at BF399310 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_068733.1 (H. sapiens) cofilin 2 (muscle) [Homo sapiens] 518 1370857_at BI282702 smooth muscle alpha-actin 519 1387018_at NM_053770 Arg/Abl-interacting protein ArgBP2 520 1398327_at BM386598 Rattus norvegicus transcribed sequence with strong similarity to protein pir: S69890 (H. sapiens) S69890 mitogen inducible gene mig-2 - human 521 1389189_at BF555956 Rattus norvegicus transcribed sequence with weak similarity to protein sp: Q9Z1P2 (R. norvegicus) AAC1_RAT Alpha-actinin 1 (Alpha-actinin cytoskeletal isoform) (Non-muscle alpha-actinin 1) (F-actin cross linking protein) 522 1371361_at BI278826 tensin 523 1375349_at BI295776 Rattus norvegicus transcribed sequence with strong similarity to protein pir: T17257 (H. sapiens) T17257 hypothetical protein DKFZp586P1422.1 - human 524 1376572_a_at AI045848 "Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_068506.1 (H. sapiens) supervillin, isoform 2; membrane-associated F-actin binding protein p205; archvillin [Homo sapiens]" 525 1367813_at NM_130403 "protein phosphatase 1, regulatory (inhibitor) subunit 14a"
526 1367648_at NM_013122 Insulin-like growth factor binding protein 2 527 1374969_at AA799832 Rattus norvegicus transcribed sequences 528 1371954_at BF290193 Rattus norvegicus transcribed sequences 529 1370347_at AF095585 enigma (LIM domain protein) 530 1388483_at BI296011 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_068733.1 (H. sapiens) cofilin 2 (muscle) [Homo sapiens] 531 1375890_at BI296994 Rattus norvegicus transcribed sequences 532 1368724_a_at NM_019131 "tropomyosin 1, alpha" 533 1370288_a_at AF372216 "tropomyosin 1, alpha" 534 1367785_at NM_031747 Calponin 1 535 1388422_at BI275904 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: JC2324 (H. sapiens) JC2324 LIM protein - human 536 1368988_at AW520914 calsequestrin 2 537 1370585_a_at X04440 "protein kinase C, beta 1" 538 1389187_at BI286421 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_115950.1 (H. sapiens) EVG1 protein [Homo sapiens] 539 1389050_at AI170797 Rattus norvegicus transcribed sequences 540 1372658_at BG373779 desmuslin 541 1387898_at D29960 heat shock 20-kDa protein 542 1373915_at AI044427 "Rattus norvegicus transcribed sequence with moderate similarity to protein pir: B49364 (H. sapiens) B49364 protein kinase (EC 2.7.1.37), myotonic dystrophy- associated - human" 543 1367570_at NM_031549 Transgelin (Smooth muscle 22 protein) 544 1370287_a_at M23764 "tropomyosin 1, alpha" 545 1386869_at NM_012893 "actin, gamma 2" 546 1388842_at BG380385 Rattus norvegicus transcribed sequences 547 1371382_at BI283060 "Rattus norvegicus transcribed sequence with strong similarity to protein pir: A37098 (H. sapiens) A37098 gelation factor ABP-280, long form - human" 548 1372219_at AA012755 "Rattus norvegicus transcribed sequence with strong similarity to protein sp: P06468 (H. sapiens) TPM2_HUMAN Tropomyosin beta chain, fibroblast and epithelial muscle-type (Tropomyosin 2, fibroblast and epithelial muscle-type) (TM36) (TME1) (TM1)" 549 1370057_at NM_017148 cysteine rich protein 1 550 1371541_at AI177055 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_444278.1 (H. sapiens) mitochondrial ribosomal protein L53 [Homo sapiens] 551 1388496_at AI103600 "Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_001449.1 (H. sapiens) gamma filamin; filamin C, gamma (actin-binding protein- 280); filamin 2; actin-binding protein 280 [Homo sapiens]" 552 1370896_a_at X16262 myosin heavy chain 11 553 1372015_at AI008689 Rattus norvegicus transcribed sequence with weak similarity to protein sp: O75410 (H. sapiens) TAC1_HUMAN Transforming acidic coiled-coil- containing protein 1 554 1372296_at AA800892 Rattus norvegicus transcribed sequence with weak similarity to protein sp: P55822 (H. sapiens) SH3B_HUMAN SH3 domain-binding glutamic acid-rich protein (SH3BGR protein) (21-glutamic acid-rich protein) (21-GARP) 555 1388451_at AA817802 Rattus norvegicus transcribed sequences 556 1388298_at BI279044 "Rattus norvegicus transcribed sequence with strong similarity to protein pir: A32031 (H. sapiens) A32031 myosin regulatory light chain, smooth muscle - human" 557 1371677_at BE113200 Rattus norvegicus transcribed sequences 558 1367691_at NM_134449 PKC-delta binding protein 559 1372159_at BI285456 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_067541.1 (M. musculus) junctophilin 2 [Mus musculus] 560 1371331_at BG665037 Rattus norvegicus transcribed sequences 561 1370234_at AA893484 Fibronectin 1 562 1372111_at BI285449 caveolin 563 1399065_at BM389543 Rattus norvegicus transcribed sequences 564 1371855_at BM390254 Rattus norvegicus transcribed sequences 565 1389394_at AI411809 Rattus norvegicus transcribed sequences 566 1368145_at NM_013002 Neuron specific protein PEP-19 (Purkinje cell protein 4) 567 1371566_at BI296437 Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_116264.1 (H. sapiens) hypothetical protein MGC15482 [Homo sapiens] 568 1368105_at AI228231 Tspan-2 protein 569 1372625_at AA851663 Rattus norvegicus transcribed sequences 570 1372537_at BI289642 Rattus norvegicus transcribed sequence with strong similarity to protein sp: O00423 (H. sapiens) EML1_HUMAN Echinoderm microtubule-associated protein-like 1 (EMAP-1) (HuEMAP-1) 571 1390430_at BF284190 "nuclear receptor subfamily 1, group D, member 2" 572 1371969_at BI291848 Rattus norvegicus transcribed sequences 573 1386860_at NM_012811 milk fat globule-EGF factor 8 protein 574 1371588_at AA686007 "parvin, alpha" 575 1387224_at NM_019304 "diacylglycerol kinase, beta" 576 1374237_at BI286025 "Rattus norvegicus transcribed sequence with strong similarity to protein sp: P29536 (H. sapiens) LMD1_HUMAN Leiomodin 1 (Leiomodin, muscle form) (64 kDa autoantigen D1) (64 kDa autoantigen 1D) (64 kDa autoantigen 1D3) (Thyroid-associated ophthalmopathy aut 577 1373911_at BM389026 Rattus norvegicus transcribed sequence with moderate similarity to protein pir: S36110 (H. sapiens) S36110 osteoblast-specific factor 2 - human 578 1371732_at BI285485 Rattus norvegicus transcribed sequences 579 1371700_at AI177059 Rattus norvegicus transcribed sequence with strong similarity to protein sp: P55083 (H. sapiens) MFA4_HUMAN Microfibril-associated glycoprotein 4 580 1376099_at AW254017 "collagen, type V, alpha 1" 581 1388935_at AI231814 Rattus norvegicus transcribed sequences 582 1379936_at AA875132 Rattus norvegicus transcribed sequences 583 1372342_at AI176583 Rattus norvegicus transcribed sequences 584 1398370_at AW522471 rexo70 585 1369652_at AI145313 Thymus cell surface antigen 586 1376640_at BF285466 Rattus norvegicus transcribed sequences 587 1369955_at NM_134452 "collagen, type V, alpha 1" 588 1373858_at BE109064 "Rattus norvegicus transcribed sequence with moderate similarity to protein pdb: 1LBG (E. coli) B Chain B, Lactose Operon Repressor Bound To 21-Base Pair Symmetric Operator Dna, Alpha Carbons Only" 589 1387854_at BI282748 "procollagen, type I, alpha 2" 590 1386862_at NM_013132 annexin 5 591 1388116_at BI285575 "collagen, type 1, alpha 1" 592 1370959_at BI275716 "collagen, type III, alpha 1" 593 1370155_at BM388837 "procollagen, type I, alpha 2" 594 1388569_at AI179984 alpha-2 antiplasmin 595 1373032_at AW251450 fracture callus protein MUSTANG 596 1376265_at AI411542 Rattus norvegicus transcribed sequences 597 1389367_at AI409747 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_055390.1 (H. sapiens) schwannomin interacting protein 1 [Homo sapiens] 598 1373957_at BF281544 Reelin 599 1370927_at BE108345 "procollagen, type XII, alpha 1" 600 1372305_at AA893634 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_057513.1 (H. sapiens) COPZ2 for nonclathrin coat protein zeta-COP [Homo sapiens] 601 1383822_at AI029990 "Rattus norvegicus transcribed sequence with weak similarity to protein ref: NP_001705.1 (H. sapiens) Bicaudal D homolog 1 (Drosophila); Bicaudal-D, Drosophila, homolog of, 1; Bicaudal D (Drosophila) homolog 1 [Homo sapiens]" 602 1374774_at BF552241 Rattus norvegicus transcribed sequences 603 1388312_at BI274487 tissue inhibitor of metalloproteinase 2 604 1376662_at BF390141 Rattus norvegicus transcribed sequences 605 1374971_at AA818954 Rattus norvegicus transcribed sequences 606 1389066_at BI274408 Rattus norvegicus transcribed sequences 607 1373102_at BI282750 Rattus norvegicus transcribed sequences 608 1388738_at AI411227 Rattus norvegicus transcribed sequences 609 1368842_at BG377130 Rattus norvegicus transcribed sequences 610 1390444_at AI044433 Rattus norvegicus transcribed sequences 611 1377087_at AA963029 Rattus norvegicus transcribed sequences 612 1387042_at NM_012828 "calcium channel, voltage-dependent, beta 3 subunit" 613 1369974_at NM_012663 vesicle-associated membrane protein 2 614 1374870_at BF286402 Rattus norvegicus transcribed sequences 615 1374087_at AI411088 Rattus norvegicus transcribed sequences 616 1374117_at BI279562 brain-specific angiogenesis inhibitor 1-associated protein 2 617 1390016_at BF415854 Rattus norvegicus transcribed sequences 618 1390341_at BF396709 Rattus norvegicus transcribed sequences 619 1374709_at AI406795 Rattus norvegicus transcribed sequences 620 1374726_at AI411941 Rattus norvegicus transcribed sequences 621 1389511_s_at BF403383 synaptogyrin 1 622 1390489_at BE108860 Rattus norvegicus transcribed sequences 623 1388598_at BI281230 Rattus norvegicus transcribed sequences 624 1370375_at J05499 liver mitochondrial glutaminase 625 1388821_at AI010430 Rattus norvegicus transcribed sequences 626 1368276_at NM_012664 Synaptophysin 627 1375862_at BM384701 Rattus norvegicus transcribed sequence with weak similarity to protein sp: P07202 (H. sapiens) PERT_HUMAN Thyroid peroxidase precursor (TPO) 628 1377136_at AW254190 Rattus norvegicus transcribed sequences 629 1392887_at AI575082 Rattus norvegicus transcribed sequences 630 1389157_at BI275583 Rattus norvegicus transcribed sequences 631 1377013_at AI639108 Rattus norvegicus transcribed sequences 632 1371389_at AI170668 Rattus norvegicus transcribed sequences 633 1370969_at BE107303 Rattus norvegicus transcribed sequence with moderate similarity to protein sp: P03845 (E. coli) YPA1_ECOLI HYPOTHETICAL PROTEIN 1 634 1373438_at BE329352 Rattus norvegicus transcribed sequence with weak similarity to protein pir: RGECDW (E. coli) RGECDW transcription activator of D-serine dehydratase - Escherichia coli 635 1388456_at AI228548 "Rattus norvegicus transcribed sequence with strong similarity to protein sp: P23297 (H. sapiens) S10A_HUMAN S-100 protein, alpha chain (S100 calcium-binding protein A1)" 636 1373427_at BI288816 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_067067.1 (H. sapiens) Rag D protein; hypothetical GTP-binding protein DKFZp761H171 [Homo sapiens] 637 1368085_at NM_133595 GTP cyclohydrolase I feedback regulatory protein 638 1369103_at NM_012755 Fyn proto-oncogene 639 1389456_at BI296591 Rattus norvegicus transcribed sequences 640 1370904_at BI301490 R. norvegicus mRNA for RT1.Ma 641 1371499_at AI227627 CD9 antigen (p24) 642 1371362_at BI285402 Rattus norvegicus transcribed sequence with strong similarity to protein sp: Q92841 (H. sapiens) DD17_HUMAN Probable RNA-dependent helicase p72 (DEAD-box protein p72) (DEAD-box protein 17) 643 1388920_at AI230985 bone morphogenetic protein 6 644 1368851_at NM_012555 v-ets erythroblastosis virus E26 oncogene homolog 1 (avian) 645 1373343_at AI175820 Rattus norvegicus transcribed sequences 646 1374888_at AI317837 Rattus norvegicus transcribed sequences 647 1376453_at BF419074 Rattus norvegicus transcribed sequences 648 1376062_at BG375315 syndecan 1 649 1371046_at AW920849 "Rattus norvegicus beta II spectrin-short isoform mRNA, partial cds" 650 1368080_at NM_054008 Rgc32 protein 651 1371931_at BI274753 general transcription factor II I 652 1388463_at AW252660 Rattus norvegicus transcribed sequence with moderate similarity to protein ref: NP_057010.1 (H. sapiens) putative secreted protein; similar to putative secreted protein (H. sapiens) [Homo sapiens] 653 1388639_at BF284148 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_060149.1 (H. sapiens) hypothetical protein FLJ20128 [Homo sapiens] 654 1379685_at AI502753 CTD-binding SR-like protein rA4 655 1389967_at AA892386 "Rattus norvegicus ADP-ribosylation-like factor 6- interacting protein mRNA, complete cds" 656 1368941_at NM_032076 prostaglandin E receptor 4 (subtype EP4) 657 1390457_at BM385762 Rattus norvegicus transcribed sequences 658 1390458_at BG666849 Rattus norvegicus transcribed sequences 659 1375254_at BE103444 Rattus norvegicus transcribed sequence with strong similarity to protein pir: JC5023 (H. sapiens) JC5023 CMP-sialic acid transporter - human 660 1375530_at BG374612 Rattus norvegicus transcribed sequence with strong similarity to protein ref: NP_062298.1 (M. musculus) glucosamine-phosphate N-acetyltransferase 1;
glucosamine-6-phosphate acetyltransferase; glucosamine- phosphate N-acetyltransferase [Mus musculus]
TABLE-US-00006 TABLE 6 Genes differentially expressed in CNI1493-treated S1 DRG No. Identifier Acce. No. CTRL-VHL CTRL-CNI IBS-VHL IBS-CNI Description 52 1373932_at BE098739 1.00 4.46 1.91 6.75 ESTs 48 1370913_at AI409634 1.00 1.70 0.98 3.44 Best5 protein 15 1369202_at NM_017028 1.00 2.20 0.35 3.04 myxovirus (influenza virus) resistance 2 62 1371152_a_at Z18877 1.00 1.49 0.48 2.48 25 oligoadenylate synthetase 53 1367850_at NM_053843 1.00 1.66 1.01 2.32 "Fc receptor, IgG, low affinity III" 54 1389006_at AI170394 1.00 1.53 1.03 2.28 ESTs 58 1383564_at BF411036 1.00 1.34 0.52 2.26 "ESTs, Highly similar to IRF7 MOUSE Interferon regulatory factor 7 (IRF-7) [M. musculus]" 67 1389034_at BI295179 1.00 1.49 0.59 2.24 "ESTs, Moderately similar to UBPI_MOUSE Ubiquitin carboxyl-terminal hydrolase 18 (Ubiquitin thiolesterase 18) (Ubiquitin-specific processing protease 18) (Deubiquitinating enzyme 18) (43 kDa ubiquitin- specific protease) [M. musculus]" 35 1386166_at AA892881 1.00 1.59 1.43 2.18 EST 69 1388164_at AF029241 1.00 1.52 0.97 2.09 "Rattus norvegicus partial mRNA for BM1k MHC class Ib antigen, strain SHR" 65 1370892_at BI285347 1.00 1.66 0.69 2.05 Complement component 4 73 1369186_at D85899 1.00 1.78 1.10 2.05 caspase 1 56 1372691_at BI292558 1.00 1.34 0.94 2.02 "ESTs, Highly similar to A57501 uridine phosphorylase (EC 2.4.2.3) I - mouse [M. musculus]" 70 1371015_at X52711 1.00 1.40 0.81 1.98 "Myxovirus (influenza) resistance, homolog of murine Mx (also interferon-inducible protein IFI78)" 74 1376151_a_at AI407953 1.00 1.51 0.92 1.95 ESTs 51 1374730_at AI102519 1.00 1.42 1.11 1.91 "ESTs, Highly similar to TYRO protein tyrosine kinase binding protein; killer cell activating receptor associated protein [Mus musculus] [M. musculus]" 30 1390730_at BM383911 1.00 1.69 1.33 1.86 ESTs 36 1375346_at BI290002 1.00 1.62 1.43 1.82 "ESTs, Weakly similar to hypothetical protein FLJ20010 [Homo sapiens] [H. sapiens]" 72 1390738_at BM385476 1.00 1.66 1.20 1.74 ESTs 47 1370186_at AI599350 1.00 1.32 1.05 1.74 "proteosome (prosome, macropain) subunit, beta type 9 (large multifunctional protease 2)" 80 1389553_at BF393825 1.00 1.16 0.85 1.73 "ESTs, Weakly similar to RIKEN cDNA 1810046124 [Mus musculus] [M. musculus]" 77 1372930_at AI411381 1.00 1.14 0.76 1.72 "ESTs, Weakly similar to DEAF-1 related transcriptional regulator (NUDR) [Rattus norvegicus] [R. norvegicus]" 59 1367786_at NM_080767 1.00 1.23 0.73 1.66 "proteasome (prosome, macropain) subunit, beta type, 8 (low molecular mass polypeptide 7)" 49 1388071_x_at M24024 1.00 1.28 1.05 1.66 RT1 class Ib gene 44 1380030_at AW523520 1.00 1.49 0.96 1.65 "ESTs, Highly similar to RIKEN cDNA 3110024A21 [Mus musculus] [M. musculus]" 57 1390510_at BI294706 1.00 1.21 0.83 1.65 "ESTs, Weakly similar to RIKEN cDNA 1810027D10 [Mus musculus] [M. musculus]" 42 1374944_at BI275829 1.00 1.26 0.65 1.65 "ESTs, Moderately similar to hypothetical protein LOC51058 [Homo sapiens] [H. sapiens]" 23 1384446_at BF402271 1.00 1.72 1.51 1.62 ESTs 34 1375988_at AW914928 1.00 1.39 1.38 1.62 ESTs 27 1373403_at AI230625 1.00 1.59 1.11 1.61 ESTs 66 1387995_a_at BI285494 1.00 1.40 0.84 1.60 interferon-inducible protein variant 10 39 1387969_at U22520 1.00 1.52 1.06 1.59 "small inducible cytokine B subfamily (Cys-X-Cys), member 10" 32 1384167_at AW522260 1.00 1.70 1.38 1.59 ESTs 37 1387566_at NM_133551 1.00 1.42 1.25 1.59 "phospholipase A2, group IVA (cytosolic, calcium- dependent)" 55 1372516_at AI317842 1.00 1.24 1.06 1.58 "ESTs, Weakly similar to S62328 kinesin-like DNA binding protein KID - human [H. sapiens]" 43 1374277_at BI289615 1.00 1.44 1.01 1.56 ESTs 28 1376264_at AW526697 1.00 1.56 1.51 1.56 ESTs 33 1388872_at BI290053 1.00 1.25 1.19 1.56 isopentenyl-diphosphate delta isomerase 14 1390312_at BG670441 1.00 1.45 0.77 1.55 "ESTs, Weakly similar to hypothetical protein FLJ20073 [Homo sapiens] [H. sapiens]" 64 1388149_at X57523 1.00 1.15 0.72 1.52 "Transporter 1, ABC (ATP binding cassette)" 71 1389571_at BG666368 1.00 1.20 0.95 1.52 "ESTs, Weakly similar to signal transducer and activator of transcription 1 [Rattus norvegicus] [R. norvegicus]" 68 1368835_at AW434718 1.00 1.33 0.87 1.51 signal transducer and activator of transcription 1 60 1388347_at AI233210 1.00 1.11 0.93 1.51 "ESTs, Moderately similar to I49013 thymic shared antigen- 1 - mouse [M. musculus]" 12 1376693_at AA998964 1.00 1.27 0.60 1.49 "ESTs, Moderately similar to hypothetical protein FLJ20073 [Homo sapiens] [H. sapiens]" 50 1373043_at BI275923 1.00 1.36 1.16 1.49 "ESTs, Highly similar to stromal cell-derived factor 2- like 1 [Mus musculus] [M. musculus]" 24 1374465_at AI237098 1.00 1.48 1.04 1.49 "ESTs, Highly similar to ubiquitously expressed transcript [Mus musculus] [M. musculus]" 63 1373025_at AI411618 1.00 1.15 0.61 1.47 "ESTs, Weakly similar to S49158 complement protein C1q beta chain precursor - rat [R. norvegicus]" 25 1369962_at NM_031014 1.00 1.58 1.36 1.47 5-aminoimidazole-4- carboxamide ribonucleotide formyltransferase/IMP 19 1377117_at BF387780 1.00 1.88 0.94 1.45 ESTs 16 1388754_at AI176839 1.00 1.49 0.88 1.45 ESTs 46 1376056_at BF291214 1.00 1.33 1.07 1.43 "ESTs, Moderately similar to hypothetical protein FLJ14464 [Homo sapiens] [H. sapiens]" 29 1399056_at AI716436 1.00 1.57 1.21 1.43 "ESTs, Moderately similar to candidate tumor suppressor protein [Homo sapiens] [H. sapiens]" 3 1376751_at AI044250 1.00 1.74 1.16 1.41 "ESTs, Highly similar to ST19_MOUSE Serine/threonine-protein kinase 19 (RP1 protein) [M. musculus]" 11 1367846_at NM_012618 1.00 1.59 0.95 1.41 S100 calcium-binding protein A4 45 1373504_at BF287967 1.00 1.32 0.96 1.40 "ESTs, Weakly similar to JE0204 testicular protein Tpx- 1 - rat [R. norvegicus]" 22 1372043_at BI282363 1.00 1.52 1.16 1.37 "ESTs, Moderately similar to ribosomal protein P0-like protein; 60S acidic ribosomal protein PO [Homo sapiens] [H. sapiens]" 38 1370214_at AI175539 1.00 1.18 0.84 1.34 Parvalbumin (calcium binding protein) 9 1390655_at BF420653 1.00 1.47 0.54 1.34 "ESTs, Highly similar to T14155 zinc finger protein Peg3 - mouse [M. musculus]" 26 1375669_at BI285619 1.00 1.39 1.03 1.32 "ESTs, Weakly similar to FK506 binding protein 2 (13 kDa) [Rattus norvegicus] [R. norvegicus]" 21 1398884_at BM384924 1.00 1.71 1.10 1.32 "ESTs, Highly similar to prefoldin 5; EIG-1; c-myc binding protein MM-1; DNA segment, Chr 15, ERATO Doi 697, expressed [Mus musculus] [M. musculus]" 1 1368465_at NM_012892 1.00 1.66 1.12 1.30 amiloride-sensitive cation channel 1 20 1371381_at AI007981 1.00 1.46 0.99 1.29 "ESTs, Moderately similar to UCRX_HUMAN Ubiquinol- cytochrome C reductase complex 7.2 kDa protein (Cytochrome C1, nonheme 7 kDa protein) (Complex III subunit X) (7.2 kDa cytochrome c1-associated protein subunit) (HSPC119) [H. sapiens]" 4 1372815_at BG673028 1.00 1.70 1.02 1.29 "ESTs, Highly similar to MGN_HUMAN Mago nashi protein homolog [M. musculus]" 41 1388628_at BI284430 1.00 1.30 0.84 1.29 "ESTs, Weakly similar to PSD7_MOUSE 26S proteasome non-ATPase regulatory subunit 7 (26S proteasome regulatory subunit S12) (Proteasome subunit p40) (Mov34 protein) [M. musculus]" 61 1373037_at BI279216 1.00 1.07 0.74 1.26 "ESTs, Weakly similar to ubiquitin-conjugating enzyme E2D 2 [Rattus norvegicus] [R. norvegicus]" 2 1374950_at BF400611 1.00 1.52 0.89 1.23 ESTs 10 1386425_at H33472 1.00 1.27 0.73 1.23 EST 31 1390065_at BE096685 1.00 1.53 0.86 1.22 ESTs 17 1390389_at BM385936 1.00 1.21 0.87 1.20 ESTs 7 1369970_at NM_031827 1.00 1.45 0.89 1.18 vesicle-associated membrane protein 8 (endobrevin) 13 1390562_s_at BE102350 1.00 1.09 0.68 1.15 ESTs 8 1390147_at AI549079 1.00 1.26 0.75 1.12 "ESTs, Highly similar to T17232 hypothetical protein DKFZp434I116.1 - human (fragment) [H. sapiens]" 95 1369157_at NM_017229 1.00 1.05 1.77 1.09 phosphodiesterase 3B 78 1387242_at NM_019335 1.00 0.96 2.62 0.95 "Protein kinase, interferon- inducible double stranded RNA dependent" 79 1377225_at BF400628 1.00 0.76 3.47 0.85 ESTs 18 1372137_at BI279854 1.00 1.13 0.73 1.03 "ESTs, Highly similar to GC5L_MOUSE GCN5-LIKE PROTEIN 1 [M. musculus]" 75 1393280_at AA874924 1.00 1.18 0.28 1.09 "ESTs, Moderately similar to lymphocyte antigen 86 [Mus musculus] [M. musculus]" 76 1372604_at BI289459 1.00 1.55 0.08 1.29 "ESTs, Weakly similar to apolipoprotein L, 3; TNF- inducible protein CG12-1 [Homo sapiens] [H. sapiens]" 102 1374802_at AI010721 1.00 0.90 1.72 0.96 "ESTs, Moderately similar to hypothetical protein FLJ20424 [Homo sapiens] [H. sapiens]" 40 1371171_at M10094 1.00 0.76 6.16 0.89 RT1 class Ib gene 88 1371725_at BM392410 1.00 0.56 1.01 0.95 ESTs 5 1368316_at NM_019158 1.00 1.22 0.71 0.91 aquaporin 8 89 1375084_at BF419780 1.00 0.64 1.12 0.90 ESTs 85 1398272_at NM_022860 1.00 0.66 0.99 0.90 beta-4N- acetylgalactosaminyltransferase 81 1374721_at AI178647 1.00 0.74 1.15 0.89 ESTs 90 1374398_at AI178787 1.00 0.68 1.10 0.86 "ESTs, Highly similar to suppressor of Ty 6 homolog (S. cerevisiae) [Mus musculus] [M. musculus]" 91 1369999_a_at NM_053601 1.00 0.68 0.95 0.84 neuronatin 6 1387658_at U93849 1.00 1.61 0.43 0.83 Eukaryotic elongation factor 2 kinase 129 1387349_at NM_013028 1.00 0.56 0.76 0.81 Short stature homeobox 2 94 1390347_at AW535909 1.00 0.72 1.02 0.80 ESTs 100 1376944_at AI407163 1.00 0.94 1.34 0.79 ESTs 96 1369052_at NM_133323 1.00 0.90 1.29 0.79 zinc finger protein 111 84 1376410_at BI291814 1.00 0.66 0.93 0.76 ESTs 118 1389193_at BM388083 1.00 0.60 0.83 0.76 ESTs 99 1376537_at AW435010 1.00 0.81 1.06 0.75 ESTs
136 1374655_at BG378095 1.00 0.57 0.90 0.75 ESTs 97 1375983_at AI234287 1.00 0.77 1.21 0.74 ESTs 92 1392890_at BG663460 1.00 0.59 1.24 0.74 ESTs 135 1369061_at NM_053906 1.00 0.25 0.81 0.74 glutathione reductase 101 1376610_a_at BE109163 1.00 0.77 1.60 0.74 "ESTs, Weakly similar to G02540 nucleobindin - human [H. sapiens]" 103 1377823_at AW531363 1.00 1.04 1.60 0.74 ESTs 82 1388116_at BI285575 1.00 0.56 1.13 0.73 "collagen, type 1, alpha 1" 121 1377070_at BE098910 1.00 0.59 0.72 0.73 "ESTs, Moderately similar to S3B2_HUMAN Splicing factor 3B subunit 2 (Spliceosome associated protein 145) (SAP 145) (SF3b150) (Pre-mRNA splicing factor SF3b 145 kDa subunit) [H. sapiens]" 131 1369958_at NM_022542 1.00 0.59 0.77 0.72 rhoB gene 130 1376288_at AI408455 1.00 0.64 0.78 0.71 ESTs 132 1391078_at BM391856 1.00 0.56 0.66 0.71 "ESTs, Moderately similar to A56284 differentiation- specific element binding protein - mouse [M. musculus]" 113 1374593_at AA799421 1.00 0.66 0.74 0.70 "ESTs, Highly similar to KPCE_RAT PROTEIN KINASE C, EPSILON TYPE (NPKC-EPSILON) [R. norvegicus]" 112 1372158_at BI295768 1.00 0.66 0.79 0.70 LRP16 protein 83 1370969_at BE107303 1.00 0.58 1.05 0.69 homeo box A5 93 1370583_s_at AY082609 1.00 0.73 0.98 0.69 P-glycoprotein/multidrug resistance 1 98 1370139_a_at AB051214 1.00 0.87 1.16 0.68 zzN/A 87 1389919_at BI296756 1.00 0.60 0.97 0.67 "ESTs, Weakly similar to actopaxin [Rattus norvegicus] [R. norvegicus]" 117 1368242_at NM_013186 1.00 0.60 0.72 0.66 "Potassium voltage gated channel, Shab-related subfamily, member 1" 106 1379747_at AA866443 1.00 0.83 0.97 0.66 ESTs 141 1390626_at BF398788 1.00 0.66 1.04 0.66 ESTs 133 1369113_at NM_019282 1.00 0.59 0.87 0.65 "cysteine knot superfamily 1, BMP antagonist 1" 137 1376320_at BF393051 1.00 0.75 1.08 0.65 ESTs 86 1387588_at NM_138890 1.00 0.47 1.13 0.65 zzN/A 148 1399138_at BG672805 1.00 0.71 0.87 0.65 ESTs 105 1398311_a_at AF313464 1.00 0.92 1.13 0.64 kinase D-interacting substance of 220 kDa 115 1375538_at AI230737 1.00 0.66 0.82 0.63 ESTs 140 1376204_at AW531412 1.00 0.71 0.94 0.63 ESTs 111 1390538_at BF414192 1.00 0.64 0.80 0.63 ESTs 151 1374909_at BI296626 1.00 0.69 0.90 0.62 "ESTs, Weakly similar to Gasz [Rattus norvegicus] [R. norvegicus]" 146 1368769_at NM_031760 1.00 0.82 1.01 0.62 "ATP-binding cassette, subfamily B (MDR/TAP), member 11" 107 1387929_at AB020504 1.00 0.76 0.82 0.61 PMF32 protein 149 1376464_at BE102505 1.00 0.79 0.79 0.60 ESTs 110 1375705_at AI103622 1.00 0.60 0.73 0.59 Guanine nucleotide-binding protein beta 1 128 1374084_at BE119993 1.00 0.68 0.64 0.59 ESTs 123 1390722_at AW531272 1.00 0.60 0.70 0.59 ESTs 122 1373008_x_at BF386649 1.00 0.44 0.57 0.58 reticulon 4 receptor 139 1376939_at BI284907 1.00 0.67 0.85 0.58 ESTs 119 1369059_at NM_053705 1.00 0.55 0.78 0.58 "transient receptor potential- related protein, ChaK" 126 1392500_at AA957990 1.00 0.59 0.67 0.58 ESTs 152 1369048_at NM_017289 1.00 0.71 0.64 0.56 "gamma-aminobutyric acid A receptor, delta" 124 1375508_at BE095963 1.00 0.50 0.61 0.56 "ESTs, Highly similar to KIF2_MOUSE KINESIN- LIKE PROTEIN KIF2 [M. musculus]" 142 1368429_at NM_133615 1.00 0.60 0.81 0.56 "TAF9-like RNA polymerase II, TATA box binding protein (TBP)-associated factor, 31 kD" 144 1373981_at BI299720 1.00 0.62 0.60 0.55 ESTs 114 1369285_at NM_031082 1.00 0.74 0.76 0.55 geranylgeranyltransferase type I (GGTase-I) 108 1372804_at AI175555 1.00 0.80 0.87 0.54 "ESTs, Highly similar to hypothetical brain protein similar to X96994 BR-1 protein (Helix pomatia) [Mus musculus] [M. musculus]" 134 1386299_at AI639471 1.00 0.50 0.86 0.53 EST 138 1377362_at BF390409 1.00 0.43 0.81 0.51 ESTs 125 1387131_at AF193015 1.00 0.44 0.55 0.50 "serine (or cysteine) proteinase inhibitor, clade I (neuroserpin), member 1" 147 1373778_at BE349670 1.00 0.83 0.92 0.46 ESTs 145 1377030_at BF390550 1.00 0.59 0.82 0.45 ESTs 143 1389436_at AI236615 1.00 0.75 0.77 0.45 ESTs 116 1386181_at AI639056 1.00 0.51 0.85 0.44 EST 150 1371042_at BG664160 1.00 0.63 0.70 0.42 mitogen-activated protein kinase kinase kinase kinase 3 109 1387453_at NM_024489 1.00 0.49 0.83 0.39 zinc finger protein RIN ZF 127 1390368_at AI716535 1.00 0.40 0.46 0.34 ESTs 104 1372640_at BI277758 1.00 0.85 1.80 0.33 ESTs 120 1369054_at NM_133518 1.00 0.48 0.60 0.25 rabphilin 3A
[0052] The human homologs of some of the rat CVHGs are provided in Table 7.
TABLE-US-00007 TABLE 7 Human homologs of rat CVHGs Nucleotide Amino acid Gene Locus ID sequence sequence desmin 1674 SEQ ID NO: 1 SEQ ID NO: 34 PEP-19 5121 SEQ ID NO: 2 SEQ ID NO: 35 IGFBP2 3485 SEQ ID NO: 3 SEQ ID NO: 36 ADAMTS1 9510 SEQ ID NO: 4 SEQ ID NO: 37 ARGBP2 8470 SEQ ID NO: 5 SEQ ID NO: 38 stathmin-like 2 11075 SEQ ID NO: 6 SEQ ID NO: 39 myxovirus resistance 2 4600 SEQ ID NO: 7 SEQ ID NO: 40 IRF7 3665 SEQ ID NO: 8 SEQ ID NO: 41 GBP2 2634 SEQ ID NO: 9 SEQ ID NO: 42 SLC28a2 9153 SEQ ID NO: 10 SEQ ID NO: 43 BDNF 627 SEQ ID NO: 11 SEQ ID NO: 44 phosphodiesterase 3B 5140 SEQ ID NO: 12 SEQ ID NO: 45 TREK2 54207 SEQ ID NO: 13 SEQ ID NO: 46 trkA 4914 SEQ ID NO: 14 SEQ ID NO: 47 IL1R1 3554 SEQ ID NO: 15 SEQ ID NO: 48 EEF2k 29904 SEQ ID NO: 16 SEQ ID NO: 49 actin, gamma 2 72 SEQ ID NO: 17 SEQ ID NO: 50 myosin heavy chain 11 4629 SEQ ID NO: 18 SEQ ID NO: 51 MRCL3 10627 SEQ ID NO: 19 SEQ ID NO: 52 MRLC2 103910 SEQ ID NO: 20 SEQ ID NO: 53 Rho family GTPase 1 27289 SEQ ID NO: 21 SEQ ID NO: 54 HSPB1 3315 SEQ ID NO: 22 SEQ ID NO: 55 RIPK4 54101 SEQ ID NO: 23 SEQ ID NO: 56 type I protein 80316 SEQ ID NO: 24 SEQ ID NO: 57 phosphatase inhibitor transgelin 6876 SEQ ID NO: 25 SEQ ID NO: 58 beta 1 integrin 3688 SEQ ID NO: 26 SEQ ID NO: 59 Desmuslin 23336 SEQ ID NO: 27 SEQ ID NO: 60 C/EBP delta 1052 SEQ ID NO: 28 SEQ ID NO: 61 FBXL22 283807 SEQ ID NO: 29 SEQ ID NO: 62 AF427491 SEQ ID NO: 30 SEQ ID NO: 63 cig5 91543 SEQ ID NO: 31 SEQ ID NO: 64 SAMD9 54809 SEQ ID NO: 32 SEQ ID NO: 65 IFI27 3429 SEQ ID NO: 33 SEQ ID NO: 66
CVHGs and CVHG Products as Markers for CVH and CVH-Related Disorders
[0053] In general, Table 3 and Table 5 provide CVHGs that are differentially expressed at in the CVH colon relative to controls. These genes may be a component in the disease mechanism and can be used as markers for diagnosing and monitoring CVH and CVH-related disorders. The CVHGs of Tables 3 and 5, as well as the corresponding CVHG products (CVHPN and CVHPP) may become novel therapeutic targets for the treatment and prevention of CVH and CVH-related disorders. Furthermore, the CVHG products themselves may be used for the treatment of CVH.
[0054] Accordingly, the present invention pertains to the use of the CVHGs listed in Tables 3 and 5, the transcribed polynucleotides (CVHPNs), and the encoded polypeptides (CVHPPs) as markers for CVH and CVH-related disorders. Moreover, the use of expression profiles of these genes can indicate the presence of or a risk of CVH and CVH-related disorders. These markers are further useful to correlate differences in levels of expression with a poor or favorable prognosis of CVH and CVH-related disorders. In particular, the present invention is directed to the use of CVHGs and panels of CVHGs set forth in Tables 3 and 5 or homologs thereof. For example, panels of the CVHGs can be conveniently arrayed on solid supports, i.e., biochips, such as the GeneChip®, for use in kits. The CVHGs can be used to assess the efficacy of a treatment or therapy of CVH and CVH-related disorders, or as a target for a treatment or therapeutic agent. The CVHGs can also be used to produce antibodies specific to CVHG products, and to construct gene therapy vectors that inhibit the development of CVH and CVH-related disorders. Therefore, without limitation as to mechanism, the invention is based in part on the principle that modulation of the expression of the CVHGs of the invention may ameliorate CVH and CVH-related disorders when they are expressed at levels similar or substantially similar to normal (non-diseased) tissue.
[0055] In one aspect, the invention provides CVHGs whose level of expression, which signifies their quantity or activity, is correlated with the presence of CVH and CVH-related disorders. In certain preferred embodiments, the invention is performed by detecting the presence of an CVHPN or a CVHPP.
[0056] In another aspect of the invention, the expression levels of the CVHGs are determined in a particular subject sample for which either diagnosis or prognosis information is desired. The level of expression of a number of CVHGs simultaneously provides an expression profile, which is essentially a "fingerprint" of the presence or activity of an CVHG or plurality of CVHGs that is unique to the state of the cell. In certain embodiments, comparison of relative levels of expression is indicative of the severity of CVH and CVH-related disorders, and as such permits for diagnostic and prognostic analysis. Moreover, by comparing relative expression profiles of CVHGs from tissue samples taken at different points in time, e.g., pre- and post-therapy and/or at different time points within a course of therapy, information regarding which genes are important in each of these stages is obtained. The identification of genes that are abnormally expressed in CVH versus normal tissue, as well as differentially expressed genes during CVH development, allows the use of this invention in a number of ways. For example, comparison of expression profiles of CVHGs at different stages of the disease progression provides a method for long-term prognosis. In another example mentioned above, the efficacy of a particular treatment regime may be evaluated, including whether a particular drug will act to improve the long-term prognosis in a particular patient.
[0057] Similarly, CVHGs listed in Tables 4, 6 and 7 can also be used as targets of CVH treatment and as markers to monitor the efficacy of CVH treatment. The gene products from CVHGs of Tables 4, 6 and 7 may also be used in the treatment of CVH and CVH-related disorders.
[0058] The discovery of these differential expression patterns for individual or panels of CVHGs allows for screening test compounds with the goal of modulating a particular expression pattern. For example, screening can be done for compounds that will convert an expression profile for a poor prognosis to one for a better prognosis. In certain embodiments, this may be done by making biochips comprising sets of the significant CVHGs, which can then be used in these screens. These methods can also be done on the protein level; that is, protein expression levels of the CVHGs can be evaluated for diagnostic and prognostic purposes or to screen test compounds. For example, in relation to these embodiments, significant CVHGs may comprise CVHGs which are determined to have modulated activity or expression in response to a therapy regime. Alternatively, the modulation of the activity or expression of a CVHG may be correlated with the diagnosis or prognosis of CVH and CVH-related disorders.
[0059] In addition, the CVHGs listed in Tables 3-8 can be administered for gene therapy purposes, including the administration of antisense nucleic acids and RNAi. The CVHG products (including CVHPPs and CVHPNs) and modulator of CVHG products (such as anti-CVHPP antibodies) can also be administered as therapeutic drugs.
[0060] For example, the CVHG desmin has significantly increased expression in CVH tissue samples, relative to control tissue samples. The presence of increased mRNA for this gene (or any other CVHGs set forth in Tables 3 and 5), and increased levels of the protein products of this gene (or any other CVHGs set forth in Tables 3 and 5) serve as markers for CVH. Accordingly, amelioration of CVH can be achieved by modulating up-regulated CVH markers, such as desmin, to normal levels.
[0061] In another embodiment of the invention, a product of CVHG, either in the form of a polynucleotide or a polypeptide, can be used as a therapeutic compound of the invention. In yet other embodiments, a modulator of CVHG expression or the activity of an CVHG product may be used as a therapeutic compound of the invention, or may be used in combination with one or more other therapeutic compositions of the invention. Formulation of such compounds into pharmaceutical compositions is described in subsections below. Administration of such a therapeutic may suppress bioactivity of CVHG product, and therefore may be used to ameliorate CVH.
Sources of CVHG Products
[0062] The CVHG products (CVHPNs and CVHPPs) of the invention may be isolated from any tissue or cell of a subject. It will be apparent to one skilled in the art that bodily fluids, such as blood or feces, may also serve as sources from which the CVHG product of the invention may be assessed. A biological sample of the invention is obtained as a blood sample, a urine or feces sample, a colon biopsy sample. A biological sample may comprise biological components such as blood plasma, serum, erythrocytes, leukocytes, blood platelets, lymphocytes, macrophages, fibroblast cells, mast cells, fat cells, neuronal cells, epithelial cells and the like. The tissue samples containing one or more of the CVHG product themselves may be useful in the methods of the invention, and one skilled in the art will be cognizant of the methods by which such samples may be conveniently obtained, stored and/or preserved.
Isolated Polynucleotides
[0063] One aspect of the invention pertains to isolated polynucleotides. Another aspect of the invention pertains to isolated polynucleotide fragments sufficient for use as hybridization probes to identify a CVHPN in a sample, as well as nucleotide fragments for use as PCR probes/primers of the amplification or mutation of the nucleic acid molecules which encode the CVHPP of the invention.
[0064] A CVHPN molecule of the present invention, e.g., a polynucleotide molecule having the nucleotide sequence of one of the CVHGs listed in Tables 3-8, or homologs thereof, or a portion thereof, can be isolated using standard molecular biology techniques and the sequence information provided herein, as well as sequence information known in the art. Using all or a portion of the polynucleotide sequence of one of the CVHGs listed Tables 3-8 (or a homolog thereof) as a hybridization probe, a CVHG of the invention or a CVHPN of the invention can be isolated using standard hybridization and cloning techniques.
[0065] A CVHPN of the invention can be amplified using cDNA, mRNA or alternatively, genomic DNA, as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques. The polynucleotide so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis. Furthermore, oligonucleotides corresponding to CVHG nucleotide sequences of the invention can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer.
[0066] Alternatively, there are numerous amplification techniques for obtaining a full length coding sequence from a partial cDNA sequence. Within such techniques, amplification is generally performed via PCR. Any of a variety of commercially available kits may be used to perform the amplification step. Primers may be designed using, for example, software well known in the art. Primers are preferably 22-30 nucleotides in length, have a GC content of at least 50% and anneal to the target sequence at temperatures of about 68° C. to 72° C. The amplified region may be sequenced as described above, and overlapping sequences assembled into a contiguous sequence.
[0067] One such amplification technique is inverse PCR, which uses restriction enzymes to generate a fragment in the known region of the gene. The fragment is then circularized by intramolecular ligation and used as a template for PCR with divergent primers derived from the known region. Within an alternative approach, sequences adjacent to a partial sequence may be retrieved by amplification with a primer to a linker sequence and a primer specific to a known region. The amplified sequences are typically subjected to a second round of amplification with the same linker primer and a second primer specific to the known region. A variation on this procedure, which employs two primers that initiate extension in opposite directions from the known sequence, is described in WO 96/38591.
[0068] Another such technique is known as "rapid amplification of cDNA ends" or RACE. This technique involves the use of an internal primer and an external primer, which hybridizes to a polyA region or vector sequence, to identify sequences that are 5' and 3' of a known sequence. Additional techniques include capture PCR (Lagerstrom et al., PCR Methods Applic. 1:11-19, 1991) and walking PCR (Parker et al., Nucl. Acids. Res. 19:3055-60, 1991). Other methods employing amplification may also be employed to obtain a full length cDNA sequence.
[0069] In certain instances, it is possible to obtain a full length cDNA sequence by analysis of sequences provided in an expressed sequence tag (EST) database, such as that available from GenBank. Searches for overlapping ESTs may generally be performed using well known programs (e.g., NCBI BLAST searches), and such ESTs may be used to generate a contiguous full length sequence. Full length DNA sequences may also be obtained by analysis of genomic fragments.
[0070] In another preferred embodiment, an isolated polynucleotide molecule of the invention comprises a polynucleotide molecule which is a complement of the nucleotide sequence of a CVHG listed in Tables 3-8, or homolog thereof, a CVHPN of the invention, or a portion of any of these nucleotide sequences. A polynucleotide molecule which is complementary to such a nucleotide sequence is one which is sufficiently complementary to the nucleotide sequence such that it can hybridize to the nucleotide sequence, thereby forming a stable duplex.
[0071] The polynucleotide molecule of the invention, moreover, can comprise only a portion of the polynucleotide sequence of a CVHG, for example, a fragment which can be used as a probe or primer. The probe/primer typically comprises a substantially purified oligonucleotide. The oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 7 or 15, preferably about 25, more preferably about 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 400 or more consecutive nucleotides of a CVHG or a CVHPN of the invention.
[0072] Probes based on the nucleotide sequence of anCVHG or anCVHPN of the invention can be used to detect transcripts or genomic sequences corresponding to the CVHG or CVHPN of the invention. In preferred embodiments, the probe comprises a label group attached thereto, e.g., the label group can be a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor. Such probes can be used as a part of a diagnostic kit for identifying cells or tissue which misexpress (e.g., over- or under-express) a CVHG, or which have greater or fewer copies of an CVHG. For example, a level of a CVHG product in a sample of cells from a subject may be determined, or the presence of mutations or deletions of a CVHG of the invention may be assessed.
[0073] The invention further encompasses polynucleotide molecules that differ from the polynucleotide sequences of the CVHGs listed in Tables 3-8 but encode the same proteins as those encoded by the genes shown in Tables 3-8 due to degeneracy of the genetic code.
[0074] The invention also specifically encompasses homologs of the CVHGs listed in Tables 3-8 of other species. Gene homologs are well understood in the art and are available using databases or search engines such as the Pubmed-Entrez database.
[0075] The invention also encompasses polynucleotide molecules which are structurally different from the molecules described above (i.e., which have a slight altered sequence), but which have substantially the same properties as the molecules above (e.g., encoded amino acid sequences, or which are changed only in non-essential amino acid residues). Such molecules include allelic variants, and are described in greater detail in subsections herein.
[0076] In addition to the nucleotide sequences of the CVHGs listed in Tables 3-8, it will be appreciated by those skilled in the art that DNA sequence polymorphisms that lead to changes in the amino acid sequences of the proteins encoded by the CVHGs listed in Tables 3-8 may exist within a population (e.g., the human population). Such genetic polymorphism in the CVHGs listed in Tables 3-8 may exist among individuals within a population due to natural allelic variation. An allele is one of a group of genes which occur alternatively at a given genetic locus. In addition it will be appreciated that DNA polymorphisms that affect RNA expression levels can also exist that may affect the overall expression level of that gene (e.g., by affecting regulation or degradation). As used herein, the phrase "allelic variant" includes a nucleotide sequence which occurs at a given locus or to a polypeptide encoded by the nucleotide sequence.
[0077] Polynucleotide molecules corresponding to natural allelic variants and homologs of the CVHGs can be isolated based on their homology to the CVHGs listed in Tables 3-8, using the cDNAs disclosed herein, or a portion thereof, as a hybridization probe according to standard hybridization techniques under stringent hybridization conditions. Polynucleotide molecules corresponding to natural allelic variants and homologs of the CVHGs of the invention can further be isolated by mapping to the same chromosome or locus as the CVHGs of the invention.
[0078] In another embodiment, an isolated polynucleotide molecule of the invention is at least 15, 20, 25, 30, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000 or more nucleotides in length and hybridizes under stringent conditions to a polynucleotide molecule corresponding to a nucleotide sequence of an CVHG of the invention. Preferably, the isolated polynucleotide molecule of the invention hybridizes under stringent conditions to the sequence of one of the CVHGs set forth in Tables 3-8, or corresponds to a naturally-occurring polynucleotide molecule.
[0079] In addition to naturally-occurring allelic variants of the CVHG of the invention that may exist in the population, the skilled artisan will further appreciate that changes can be introduced by mutation into the nucleotide sequences of the CVHGs of the invention, thereby leading to changes in the amino acid sequence of the encoded proteins, without altering the functional activity of these proteins. For example, nucleotide substitutions leading to amino acid substitutions at "non-essential" amino acid residues can be made. A "non-essential" amino acid residue is a residue that can be altered from the wild-type sequence of a protein without altering the biological activity, whereas an "essential" amino acid residue is required for biological activity. For example, amino acid residues that are conserved among allelic variants or homologs of a gene (e.g., among homologs of a gene from different species) are predicted to be particularly unamenable to alteration.
[0080] In yet other aspects of the invention, polynucleotides of a CVHG may comprise one or more mutations. An isolated polynucleotide molecule encoding a protein with a mutation in a CVHPP of the invention can be created by introducing one or more nucleotide substitutions, additions or deletions into the nucleotide sequence of the gene encoding the CVHPP, such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein. Such techniques are well known in the art. Mutations can be introduced into the CVHG of the invention by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Preferably, conservative amino acid substitutions are made at one or more predicted non-essential amino acid residues. Alternatively, mutations can be introduced randomly along all or part of a coding sequence of a CVHG of the invention, such as by saturation mutagenesis, and the resultant mutants can be screened for biological activity to identify mutants that retain activity. Following mutagenesis, the encoded protein can be expressed recombinantly and the activity of the protein can be determined.
[0081] A polynucleotide may be further modified to increase stability in vivo. Possible modifications include, but are not limited to, the addition of flanking sequences at the 5' and/or 3' ends; the use of phosphorothioate or 2 O-methyl rather than phosphodiesterase linkages in the backbone; and/or the inclusion of nontraditional bases such as inosine, queosine and wybutosine, as well as acetyl- methyl-, thio- and other modified forms of adenine, cytidine, guanine, thymine and uridine.
[0082] Another aspect of the invention pertains to isolated polynucleotide molecules, which are antisense to the CVHGs of the invention. An "antisense" polynucleotide comprises a nucleotide sequence which is complementary to a "sense" polynucleotide encoding a protein, e.g., complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence. Accordingly, an antisense polynucleotide can hydrogen bond to a sense polynucleotide. The antisense polynucleotide can be complementary to an entire coding strand of a gene of the invention or to only a portion thereof. In one embodiment, an antisense polynucleotide molecule is antisense to a "coding region" of the coding strand of a nucleotide sequence of the invention. The term "coding region" includes the region of the nucleotide sequence comprising codons which are translated into amino acids. In another embodiment, the antisense polynucleotide molecule is antisense to a "noncoding region" of the coding strand of a nucleotide sequence of the invention.
[0083] Antisense polynucleotides of the invention can be designed according to the rules of Watson and Crick base pairing. The antisense polynucleotide molecule can be complementary to the entire coding region of an mRNA corresponding to a gene of the invention, but more preferably is an oligonucleotide which is antisense to only a portion of the coding or noncoding region. An antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 nucleotides in length. An antisense polynucleotide of the invention can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense polynucleotide can be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense polynucleotides, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used. Examples of modified nucleotides which can be used to generate the antisense polynucleotide include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxymethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenosine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense polynucleotide can be produced biologically using an expression vector into which a polynucleotide has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted polynucleotide will be of an antisense orientation to a target polynucleotide of interest, described further in the following subsection).
[0084] The antisense polynucleotide molecules of the invention are typically administered to a subject or generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding a CVHPP of the invention to thereby inhibit expression of the protein, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarity to form a stable duplex or, for example, in the cases of an antisense polynucleotide molecule which binds to DNA duplexes, through specific interactions in the major groove of the double helix. An example of a route of administration of antisense polynucleotide molecules of the invention include direct injection at a tissue site. Alternatively, antisense polynucleotide molecules can be modified to target selected cells and then administered systemically. For example, for systemic administration, antisense molecules can be modified such that they specifically bind to receptors or antigens expressed on a selected cell surface, e.g., by linking the antisense polynucleotide molecules to peptides or antibodies which bind to cell surface receptors or antigens. The antisense polynucleotide molecules can also be delivered to cells using the vectors described herein. To achieve sufficient intracellular concentrations of the antisense molecules, vector constructs comprising the antisense polynucleotide molecules are preferably placed under the control of a strong promoter.
[0085] In yet another embodiment, the antisense polynucleotide molecule of the invention is an α-anomeric polynucleotide molecule. An α-anomeric polynucleotide molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual β-units, the strands run parallel to each other. The antisense polynucleotide molecule can also comprise a 2'-o-methylribonucleotide or a chimeric RNA-DNA analogue.
[0086] In still another embodiment, an antisense polynucleotide of the invention is a ribozyme. Ribozymes are catalytic RNA molecules with ribonuclease activity which are capable of cleaving a single-stranded polynucleotide, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes) can be used to catalytically cleave mRNA transcripts of the CVHGs of the invention to thereby inhibit translation of the mRNA. A ribozyme having specificity for a CVHPN can be designed based upon the nucleotide sequence of the CVHPN. For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in a CVHG mRNA. Alternatively, mRNA transcribed from a CVHG can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. Alternatively, expression of a CVHG of the invention can be inhibited by targeting nucleotide sequences complementary to the regulatory region of the CVHG (e.g., the promoter and/or enhancers) to form triple helical structures that prevent transcription of the gene in target cells.
[0087] Expression of the CVHGs of the invention can also be inhibited using RNA interference ("RNAi"). This is a technique for post-transcriptional gene silencing ("PTGS"), in which target gene activity is specifically abolished with cognate double-stranded RNA ("dsRNA"). RNAi resembles in many aspects PTGS in plants and has been detected in many invertebrates including the trypanosome, hydra, planaria, nematode and fruit fly (Drosophila melanogaster). It may be involved in the modulation of transposable element mobilization and antiviral state formation. RNAi technology is disclosed, for example, in U.S. Pat. No. 5,919,619 and PCT Publication Nos. WO99/14346 and WO01/29058. Basically, dsRNA of about 21 nucleotides, homologous to the target gene, is introduced into the cell and a sequence specific reduction in gene activity is observed.
[0088] In yet another embodiment, the polynucleotide molecules of the present invention can be modified at the base moiety, sugar moiety or phosphate backbone to improve the stability, hybridization, or solubility of the molecule. For example, the deoxyribose phosphate backbone of the polynucleotide molecules can be modified to generate peptide polynucleotides. As used herein, the terms "peptide polynucleotides" or "PNAs" refer to polynucleotide mimics, e.g., DNA mimics, in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs has been shown to allow for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols.
[0089] PNAs can be used in therapeutic and diagnostic applications. For example, PNAs can be used as antisense agents for sequence-specific modulation of CVHG expression by, for example, inducing transcription or translation arrest or inhibiting replication. PNAs of the polynucleotide molecules of the invention can be used in the analysis of single base pair mutations in a gene, (e.g., by PNA-directed PCR clamping). They may also serve as artificial restriction enzymes when used in combination with other enzymes (e.g., S1 nucleases) or as probes or primers for DNA sequencing or hybridization.
[0090] In another embodiment, PNAs can be modified, (e.g., to enhance their stability or cellular uptake), by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras of the polynucleotide molecules of the invention can be generated which may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes, (e.g., DNA polymerases), to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation. The synthesis of PNA-DNA chimeras can be performed. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry. Modified nucleoside analogs, such as 5'-(4-methoxytrityl)amino-5'-deoxy-thymidine phosphoramidite, can be used as a spacer between the PNA and the 5' end of DNA. PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5' PNA segment and a 3' DNA segment. Alternatively, chimeric molecules can be synthesized with a 5' DNA segment and a 3' PNA segment.
[0091] In other embodiments, the oligonucleotide may include other appended groups such as peptides (e.g., for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane or the blood-kidney barrier (see, e.g. PCT Publication No. W089/10134). In addition, oligonucleotides can be modified with hybridization-triggered cleavage agents or intercalating agents. To this end, the oligonucleotide may be conjugated to another molecule (e.g., a peptide, hybridization triggered cross-linking agent, transport agent, or hybridization-triggered cleavage agent). Finally, the oligonucleotide may be detectably labeled, either such that the label is detected by the addition of another reagent (e.g., a substrate for an enzymatic label), or is detectable immediately upon hybridization of the nucleotide (e.g., a radioactive label or a fluorescent label).
Isolated Polypeptides
[0092] Several aspects of the invention pertain to isolated CVHPPs, and biologically active portions thereof, as well as polypeptide fragments suitable for use as immunogens to raise anti-CVHPP antibodies. In one embodiment, native CVHPPs can be isolated from cells or tissue sources by an appropriate purification scheme using standard protein purification techniques. Standard purification methods include electrophoretic, molecular, immunological and chromatographic techniques, including ion exchange, hydrophobic, affinity, and reverse-phase HPLC chromatography, and chromatofocusing. For example, a CVHPP may be purified using a standard anti-CVHPP antibody column. Ultrafiltration and diafiltration techniques, in conjunction with protein concentration, are also useful. The degree of purification necessary will vary depending on the use of the CVHPP. In some instances no purification will be necessary.
[0093] In another embodiment, CVHPPs or mutated CVHPPs are produced by recombinant DNA techniques. Alternative to recombinant expression, a CVHPP or mutated CVHPP can be synthesized chemically using standard peptide synthesis techniques.
[0094] The invention also provides variants of CVHPPs. The variant of a CVHPP is substantially homologous to the native CVHPP encoded by an CVHG listed in Table 4, and retains the functional activity of the native CVHPP, yet differs in amino acid sequence due to natural allelic variation or mutagenesis, as described in detail above. Accordingly, in another embodiment, the variant of a CVHPP is a protein which comprises an amino acid sequence at least about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or more homologous to the amino acid sequence of the original CVHPP.
[0095] In a non-limiting example, as used herein, proteins are referred to as "homologs" and "homologous" where a first protein region and a second protein region are compared in terms of identity. To determine the percent identity of two amino acid sequences or of two polynucleotide sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or polynucleotide sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). In a preferred embodiment, the length of a reference sequence aligned for comparison purposes is at least 30%, preferably at least 40%, more preferably at least 50%, even more preferably at least 60%, and even more preferably at least 70%, 80%, or 90% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleotide "identity" is equivalent to amino acid or nucleotide "homology"). The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps and the length of each gap, which need to be introduced for optimal alignment of the two sequences.
[0096] The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch (J. Mol. Biol. 48:444-453, 1970) algorithm which has been incorporated into the GAP program in the GCG software package, using either a Blossom 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. In yet another preferred embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package, using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6.
[0097] The polynucleotide and protein sequences of the present invention can further be used as a "query sequence" to perform a search against public databases to, for example, identify other family members or related sequences. Such searches can be performed using BLAST programs available at the BLAST website maintained by the National Center of biotechnology Information (NCBI), National Library of Medicine, Washington D.C. USA.
[0098] The invention also provides chimeric or fusion CVHPPs. Within a fusion CVHPP the polypeptide can correspond to all or a portion of a CVHPP. In a preferred embodiment, a fusion CVHPP comprises at least one biologically active portion of a CVHPP. Within the fusion protein, the term "operatively linked" is intended to indicate that the CVHPP-related polypeptide and the non-CVHPP-related polypeptide are fused in-frame to each other. The non-CVHPP-related polypeptide can be fused to the N-terminus or C-terminus of the CVHPP-related polypeptide.
[0099] A peptide linker sequence may be employed to separate the CVHPP-related polypeptide from non-CVHPP-related polypeptide components by a distance sufficient to ensure that each polypeptide folds into its secondary and tertiary structures. Such a peptide linker sequence is incorporated into the fusion protein using standard techniques well known in the art. Suitable peptide linker sequences may be chosen based on the following factors: (1) their ability to adopt a flexible extended conformation; (2) their inability to adopt a secondary structure that could interact with functional epitopes on the CVHPP-related polypeptide and non-CVHPP-related polypeptide; and (3) the lack of hydrophobic or charged residues that might react with the polypeptide functional epitopes. Preferred peptide linker sequences contain gly, asn and ser residues. Other near neutral amino acids, such as thr and ala may also be used in the linker sequence. Amino acid sequences which may be used as linkers are well known in the art. The linker sequence may generally be from 1 to about 50 amino acids in length. Linker sequences are not required when the CVHPP-related polypeptide and non-CVHPP-related polypeptide have non-essential N-terminal amino acid regions that can be used to separate the functional domains and prevent steric interference.
[0100] For example, in one embodiment, the fusion protein is a glutathione S-transferase (GST)-CVHPP fusion protein in which the CVHPP sequences are fused to the C-terminus of the GST sequences. Such fusion proteins can facilitate the purification of recombinant CVHPPs.
[0101] The CVHPP-fusion proteins of the invention can be incorporated into pharmaceutical compositions and administered to a subject in vivo, as described herein. The CVHPP-fusion proteins can be used to affect the bioavailability of a CVHPP substrate. CVHPP-fusion proteins may be useful therapeutically for the treatment of, or prevention of, damages caused by, for example, (i) aberrant modification or mutation of a CVHG; (ii) mis-regulation of a CVHG; and (iii) aberrant post-translational modification of a CVHPP.
[0102] Moreover, the CVHPP-fusion proteins of the invention can be used as immunogens to produce anti-CVHPP antibodies in a subject, to purify CVHPP ligands, and to identify molecules which inhibit the interaction of a CVHPP with a CVHPP substrate in screening assays.
[0103] Preferably, a CVHPP-chimeric or fusion protein of the invention is produced by standard recombinant DNA techniques. For example, DNA fragments coding for the different polypeptide sequences are ligated together in-frame in accordance with conventional techniques. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers which give rise to complementary overhangs between two consecutive gene fragments which can subsequently be annealed and reamplified to generate a chimeric gene sequence. Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST polypeptide). A CVHPP-encoding polynucleotide can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the CVHPP.
[0104] A signal sequence can be used to facilitate secretion and isolation of the secreted protein or other proteins of interest. Signal sequences are typically characterized by a core of hydrophobic amino acids which are generally cleaved from the mature protein during secretion in one or more cleavage events. Such signal peptides contain processing sites that allow cleavage of the signal sequence from the mature proteins as they pass through the secretory pathway. Thus, the invention pertains to the described polypeptides having a signal sequence, as well as to polypeptides from which the signal sequence has been proteolytically cleaved (i.e., the cleavage products). In one embodiment, a polynucleotide sequence encoding a signal sequence can be operably linked in an expression vector to a protein of interest, such as a protein which is ordinarily not secreted or is otherwise difficult to isolate. The signal sequence directs secretion of the protein, such as from a eukaryotic host into which the expression vector is transformed, and the signal sequence is subsequently or concurrently cleaved. The protein can then be readily purified from the extracellular medium by art recognized methods. Alternatively, the signal sequence can be linked to the protein of interest using a sequence which facilitates purification, such as with a GST domain.
[0105] The present invention also pertains to variants of the CVHPPs of the invention which function as either agonists or as antagonists to the CVHPPs. In one embodiment, antagonists or agonists of CVHPPs are used as therapeutic agents. For example, antagonists of an up-regulated CVHG that can decrease the activity or expression of such a gene and therefore ameliorate CVH in a subject wherein the CVHG is abnormally increased in level or activity. In this embodiment, treatment of such a subject may comprise administering an antagonist wherein the antagonist provides decreased activity or expression of the targeted CVHG.
[0106] In certain embodiments, an agonist of the CVHPPs can retain substantially the same, or a subset, of the biological activities of the naturally occurring form of a CVHPP or may enhance an activity of a CVHPP. In certain embodiments, an antagonist of a CVHPP can inhibit one or more of the activities of the naturally occurring form of the CVHPP by, for example, competitively modulating an activity of a CVHPP. Thus, specific biological effects can be elicited by treatment with a variant of limited function. In one embodiment, treatment of a subject with a variant having a subset of the biological activities of the naturally occurring forth of the protein has fewer side effects in a subject relative to treatment with the naturally occurring form of the CVHPP.
[0107] Mutants of a CVHPP which function as either CVHPP agonists or as CVHPP antagonists can be identified by screening combinatorial libraries of mutants, e.g., truncation mutants, of a CVHPP for CVHPP agonist or antagonist activity. In certain embodiments, such mutants may be used, for example, as a therapeutic protein of the invention. A diverse library of CVHPP mutants can be produced by, for example, enzymatically ligating a mixture of synthetic oligonucleotides into gene sequences such that a degenerate set of potential CVHPP sequences is expressible as individual polypeptides, or alternatively, as a set of larger fusion proteins (e.g., for phage display) containing the set of CVHPP sequences therein. There are a variety of methods which can be used to produce libraries of potential CVHPP variants from a degenerate oligonucleotide sequence. Chemical synthesis of a degenerate gene sequence can be performed in an automatic DNA synthesizer, and the synthetic gene is then ligated into an appropriate expression vector. Use of a degenerate set of genes allows for the provision, in one mixture, of all of the sequences encoding the desired set of potential CVHPP sequences. Methods for synthesizing degenerate oligonucleotides are known in the art.
[0108] In addition, libraries of fragments of a protein coding sequence corresponding to a CVHPP of the invention can be used to generate a diverse or heterogenous population of CVHPP fragments for screening and subsequent selection of variants of a CVHPP. In one embodiment, a library of coding sequence fragments can be generated by treating a double-stranded PCR fragment of a CVHPP coding sequence with a nuclease under conditions wherein nicking occurs only about once per molecule, denaturing the double-stranded DNA, renaturing the DNA to form double-stranded DNA which can include sense/antisense pairs from different nicked products, removing single-stranded portions from reformed duplexes by treatment with S1 nuclease, and ligating the resulting fragment library into an expression vector. By this method, an expression library can be derived which encodes N-terminal, C-terminal and internal fragments of various sizes of the CVHPP.
[0109] Several techniques are known in the art for screening gene products of combinatorial libraries made by point mutations or truncation, and for screening cDNA libraries for gene products having a selected property. The most widely used techniques, which are amenable to high-throughput analysis, for screening large gene libraries typically include cloning the gene library into replicable expression vectors, transforming appropriate cells with the resulting library of vectors, and expressing the combinatorial genes under conditions in which detection of a desired activity facilitates isolation of the vector encoding the gene whose product was detected. Recursive ensemble mutagenesis (REM), a technique which enhances the frequency of functional mutants in the libraries, can be used in combination with the screening assays to identify CVHPP variants (Delgrave et al. Protein Engineering 6:327-331, 1993).
[0110] Portions of a CVHPP or variants of a CVHPP having less than about 100 amino acids, and generally less than about 50 amino acids, may also be generated by synthetic means, using techniques well known to those of ordinary skill in the art. For example, such polypeptides may be synthesized using any of the commercially available solid-phase techniques, such as the Merrifield solid-phase synthesis method, where amino acids are sequentially added to a growing amino acid chain. Equipment for automated synthesis of polypeptides is commercially available from suppliers such as Perkin Elmer/Applied BioSystems Division (Foster City, Calif.), and may be operated according to the manufacturer's instructions.
[0111] Methods and compositions for screening for protein inhibitors or activators are known in the art (see U.S. Pat. Nos. 4,980,281, 5,266,464, 5,688,635, and 5,877,007, which are incorporated herein by reference).
[0112] It is contemplated in the present invention that CVHPPs are cleaved into fragments for use in further structural or functional analysis, or in the generation of reagents such as CVHPP and CVHPP-specific antibodies. This can be accomplished by treating purified or unpurified polypeptide with a proteolytic enzyme (i.e., a proteinase) including, but not limited to, serine proteinases (e.g., chymotrypsin, trypsin, plasmin, elastase, thrombin, substilin) metal proteinases (e.g., carboxypeptidase A, carboxypeptidase B, leucine aminopeptidase, thermolysin, collagenase), thiol proteinases (e.g., papain, bromelain, Streptococcal proteinase, clostripain) and/or acid proteinases (e.g., pepsin, gastricsin, trypsinogen). Polypeptide fragments are also generated using chemical means such as treatment of the polypeptide with cyanogen bromide (CNBr), 2-nitro-5-thiocyanobenzoic acid, isobenzoic acid, BNPA-skatole, hydroxylamine or a dilute acid solution. Recombinant techniques are also used to produce specific fragments of a CVHPP.
[0113] In addition, the invention also contemplates that compounds sterically similar to a particular CVHPP may be formulated to mimic the key portions of the peptide structure, called peptidomimetics or peptide mimetics. Mimetics are peptide-containing molecules which mimic elements of polypeptide secondary structure. See, for example, U.S. Pat. No. 5,817,879 (incorporated by reference hereinafter in its entirety). The underlying rationale behind the use of peptide mimetics is that the peptide backbone of polypeptides exists chiefly to orient amino acid side chains in such a way as to facilitate molecular interactions, such as those of receptor and ligand. Recently, peptide and glycoprotein mimetic antigens have been described which elicit protective antibody to Neisseria meningitidis serogroup B, thereby demonstrating the utility of mimetic applications (Moe et al., Int. Rev. Immunol. 20:201-20, 2001; Berezin et al., J Mol Neurosci. 22:33-39, 2004). Successful applications of the peptide mimetic concept have thus far focused on mimetics of β-turns within polypeptides. Likely β-turn structures within a CVHPP can be predicted by computer-based algorithms. For example, U.S. Pat. No. 5,933,819, incorporated by reference hereinafter in its entirety, describes a neural network based method and system for identifying relative peptide binding motifs from limited experimental data. In particular, an artificial neural network (ANN) is trained with peptides with known sequence and function (i.e., binding strength) identified from a phage display library. The ANN is then challenged with unknown peptides, and predicts relative binding motifs. Analysis of the unknown peptides validate the predictive capability of the ANN. Once the component amino acids of the turn are determined, mimetics can be constructed to achieve a similar spatial orientation of the essential elements of the amino acid side chains, as discussed in U.S. Pat. No. 6,420,119 and U.S. Pat. No. 5,817,879, and in Kyte and Doolittle, J. Mol. Biol., 157:105-132, 1982; Moe and Granoff, Int. Rev. Immunol., 20(2):201-20, 2001; Granoff et al., J. Immunol., 167(11):6487-96, 2001 (each incorporated by reference hereinafter in its entirety).
Antibodies
[0114] In another aspect, the invention includes antibodies that are specific to CVHPPs of the invention or their variants. Preferably the antibodies are monoclonal, and most preferably, the antibodies are humanized, as per the description of antibodies described below.
[0115] An isolated CVHPP, or a portion or fragment thereof, can be used as an immunogen to generate antibodies that bind the CVHPP using standard techniques for polyclonal and monoclonal antibody preparation. A full-length CVHPP can be used or, alternatively, the invention provides antigenic peptide fragments of the CVHPP for use as immunogens. The antigenic peptide of a CVHPP comprises at least 8 amino acid residues of an amino acid sequence encoded by an CVHG set forth in Tables 3-8 or an homolog thereof, and encompasses an epitope of a CVHPP such that an antibody raised against the peptide forms a specific immune complex with the CVHPP. Preferably, the antigenic peptide comprises at least 8 amino acid residues, more preferably at least 12 amino acid residues, even more preferably at least 16 amino acid residues, and most preferably at least 20 amino acid residues.
[0116] Immunogenic portions (epitopes) may generally be identified using well known techniques. Such techniques include screening polypeptides for the ability to react with antigen-specific antibodies, antisera and/or T-cell lines or clones. As used herein, antisera and antibodies are "antigen-specific" if they bind to an antigen with a binding affinity equal to, or greater than 105 M-1. Such antisera and antibodies may be prepared as described herein, and using well known techniques. An epitope of a CVHPP is a portion that reacts with such antisera and/or T-cells at a level that is not substantially less than the reactivity of the full length polypeptide (e.g., in an ELISA and/or T-cell reactivity assay). Such epitopes may react within such assays at a level that is similar to or greater than the reactivity of the full length polypeptide. Such screens may generally be performed using methods well known to those of ordinary skill in the art. For example, a polypeptide may be immobilized on a solid support and contacted with patient sera to allow binding of antibodies within the sera to the immobilized polypeptide. Unbound sera may then be removed and bound antibodies detected using, for example, 125I-labeled Protein A.
[0117] Preferred epitopes encompassed by the antigenic peptide are regions of the CVHPP that are located on the surface of the protein, e.g., hydrophilic regions, as well as regions with high antigenicity.
[0118] A CVHPP immunogen typically is used to prepare antibodies by immunizing a suitable subject, (e.g., rabbit, goat, mouse or other mammal) with the immunogen. An appropriate immunogenic preparation can contain, for example, recombinantly expressed CVHPP or a chemically synthesized CVHPP. The preparation can further include an adjuvant, such as Freund's complete or incomplete adjuvant, or a similar immunostimulatory agent. Immunization of a suitable subject with an immunogenic CVHPP preparation induces a polyclonal anti-CVHPP antibody response. Techniques for preparing, isolating and using antibodies are well known in the art.
[0119] Accordingly, another aspect of the invention pertains to monoclonal or polyclonal anti-CVHPP antibodies and immunologically active portions of the antibody molecules, including F(ab) and F(ab')2 fragments which can be generated by treating the antibody with an enzyme such as pepsin.
[0120] Polyclonal anti-CVHPP antibodies can be prepared as described above by immunizing a suitable subject with a CVHPP. The anti-CVHPP antibody titer in the immunized subject can be monitored over time by standard techniques, such as with an enzyme linked immunosorbent assay (ELISA) using immobilized CVHPP. If desired, the antibody molecules directed against CVHPPs can be isolated from the subject (e.g., from the blood) and further purified by well known techniques, such as protein A chromatography, to obtain the IgG fraction. At an appropriate time after immunization, e.g., when the anti-CVHPP antibody titers are highest, antibody-producing cells can be obtained from the subject and used to prepare monoclonal antibodies by standard techniques, such as the hybridoma technique, human B cell hybridoma technique, the EBV-hybridoma technique, or trioma techniques. The technology for producing monoclonal antibody hybridomas is well known. Briefly, an immortal cell line (typically a myeloma) is fused to lymphocytes (typically splenocytes) from a mammal immunized with a CVHPP immunogen as described above, and the culture supernatants of the resulting hybridoma cells are screened to identify a hybridoma producing a monoclonal antibody that binds to a CVHPP of the invention.
[0121] Any of the many well known protocols used for fusing lymphocytes and immortalized cell lines can be applied for the purpose of generating an anti-CVHPP monoclonal antibody. Moreover, the ordinarily skilled worker will appreciate that there are many variations of such methods which also would be useful. Typically, the immortal cell line (e.g., a myeloma cell line) is derived from the same mammalian species as the lymphocytes. For example, murine hybridomas can be made by fusing lymphocytes from a mouse immunized with an immunogenic preparation of the present invention with an immortalized mouse cell line. Preferred immortal cell lines are mouse myeloma cell lines that are sensitive to culture medium containing hypoxanthine, aminopterin and thymidine ("HAT medium"). Any of a number of myeloma cell lines can be used as a fusion partner according to standard techniques, e.g., the P3-NS1/1-Ag4-1, P3-x63-Ag8.653 or Sp210-Ag14 myeloma lines. These myeloma lines are available from ATCC. Typically, HAT-sensitive mouse myeloma cells are fused to mouse splenocytes using polyethylene glycol ("PEG"). Hybridoma cells resulting from the fusion are then selected using HAT medium, which kills unfused and unproductively fused myeloma cells (unfused splenocytes die after several days because they are not transformed). Hybridoma cells producing a monoclonal antibody of the invention are detected by screening the hybridoma culture supernatants for antibodies that bind to a CVHPP, e.g., using a standard ELISA assay.
[0122] Alternative to preparing monoclonal antibody-secreting hybridomas, a monoclonal anti-CVHPP antibody can be identified and isolated by screening a recombinant combinatorial immunoglobulin library (e.g., an antibody phase display library) with CVHPP to thereby isolate immunoglobulin library members that bind to a CVHPP. Kits for generating and screening phage display libraries are commercially available.
[0123] The anti-CVHPP antibodies also include "Single-chain Fv" or "scFv" antibody fragments. The scFv fragments comprise the VH and VL domains of antibody, wherein these domains are present in a single polypeptide chain. Generally, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen binding.
[0124] Additionally, recombinant anti-CVHPP antibodies, such as chimeric and humanized monoclonal antibodies, comprising both human and non-human portions, which can be made using standard recombinant DNA techniques, are within the scope of the invention. Such chimeric and humanized monoclonal antibodies can be produced by recombinant DNA techniques known in the art.
[0125] Humanized antibodies are particularly desirable for therapeutic treatment of human subjects. Humanized forms of non-human (e.g., murine) antibodies are chimeric molecules of immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab')2 or other antigen-binding subsequences of antibodies), which contain minimal sequence derived from non-human immunoglobulin. Humanized antibodies include human immunoglobulins (recipient antibody) in which residues forming a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity. In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies may also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the constant regions being those of a human immunoglobulin consensus sequence. The humanized antibody will preferably also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.
[0126] Such humanized antibodies can be produced using transgenic mice which are incapable of expressing endogenous immunoglobulin heavy and light chain genes, but which can express human heavy and light chain genes. The transgenic mice are immunized in the normal fashion with a selected antigen, e.g., all or a portion of a polypeptide corresponding to a CVHPP of the invention. Monoclonal antibodies directed against the antigen can be obtained using conventional hybridoma technology. The human immunoglobulin transgenes harbored by the transgenic mice rearrange during B cell differentiation, and subsequently undergo class switching and somatic mutation. Thus, using such a technique, it is possible to produce therapeutically useful IgG, IgA and IgE antibodies.
[0127] Humanized antibodies which recognize a selected epitope can be generated using a technique referred to as "guided selection." In this approach a selected non-human monoclonal antibody, e.g., a murine antibody, is used to guide the selection of a humanized antibody recognizing the same epitope.
[0128] In a preferred embodiment, the antibodies to CVHPP are capable of reducing or eliminating the biological function of CVHPP, as is described below. That is, the addition of anti-CVHPP antibodies (either polyclonal or preferably monoclonal) to CVHPP (or cells containing CVHPP) may reduce or eliminate the CVHPP activity. Generally, at least a 25% decrease in activity is preferred, with at least about 50% being particularly preferred and about a 95-100% decrease being especially preferred.
[0129] An anti-CVHPP antibody can be used to isolate a CVHPP of the invention by standard techniques, such as affinity chromatography or immunoprecipitation. An anti-CVHPP antibody can facilitate the purification of natural CVHPPs from cells and of recombinantly produced CVHPPs expressed in host cells. Moreover, an anti-CVHPP antibody can be used to detect a CVHPP (e.g., in a cellular lysate or cell supernatant on the cell surface) in order to evaluate the abundance and pattern of expression of the CVHPP. Anti-CVHPP antibodies can be used diagnostically to monitor protein levels in tissue as part of a clinical testing procedure, for example, to determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling (i.e., physically linking) the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin; and examples of suitable radioactive materials include 125I, 131I, 35S or 3H.
[0130] Anti-CVHPP antibodies of the invention are also useful for targeting a therapeutic to a cell or tissue comprising the antigen of the anti-CVHPP antibody. For example, a therapeutic, such as a small molecule, can be linked to the anti-CVHPP antibody in order to target the therapeutic to the cell or tissue comprising the CVHPP antigen. The method is particularly useful in connection with CVHPPs which are surface markers.
[0131] A therapeutic agent may be coupled (e.g., covalently bonded) to a suitable monoclonal antibody either directly or indirectly (e.g., via a linker group). A direct reaction between an agent and an antibody is possible when each possesses a substituent capable of reacting with the other. For example, a nucleophilic group, such as an amino or sulfhydryl group, on one may be capable of reacting with a carbonyl-containing group, such as an anhydride or an acid halide, or with an alkyl group containing a good leaving group (e.g., a halide) on the other.
[0132] Alternatively, it may be desirable to couple a therapeutic agent and an antibody via a linker group. A linker group can function as a spacer to distance an antibody from an agent in order to avoid interference with binding capabilities. A linker group can also serve to increase the chemical reactivity of a substituent on an agent or an antibody, and thus increase the coupling efficiency. An increase in chemical reactivity may also facilitate the use of agents, or functional groups on agents, which otherwise would not be possible.
[0133] It will be evident to those skilled in the art that a variety of bifunctional or polyfunctional reagents, both homo- and hetero-functional, may be employed as the linker group. Coupling may be effected, for example, through amino groups, carboxyl groups, sulfhydryl groups or oxidized carbohydrate residues. There are numerous references describing such methodology, e.g., U.S. Pat. No. 4,671,958, to Rodwell et al.
[0134] Where a therapeutic agent is more potent when free from the antibody portion of the immunoconjugates of the present invention, it may be desirable to use a linker group which is cleavable during or upon internalization into a cell. A number of different cleavable linker groups have been described. The mechanisms for the intracellular release of an agent from these linker groups include cleavage by reduction of a disulfide bond (e.g., U.S. Pat. No. 4,489,710, to Spitler), by irradiation of a photolabile bond (e.g., U.S. Pat. No. 4,625,014, to Senter et al.), by hydrolysis of derivatized amino acid side chains (e.g., U.S. Pat. No. 4,638,045, to Kohn et al.), by serum complement-mediated hydrolysis (e.g., U.S. Pat. No. 4,671,958, to Rodwell et al.), and acid-catalyzed hydrolysis (e.g., U.S. Pat. No. 4,569,789, to Blattler et al.).
[0135] It may be desirable to couple more than one agent to an antibody. In one embodiment, multiple molecules of an agent are coupled to one antibody molecule. In another embodiment, more than one type of agent may be coupled to one antibody. Regardless of the particular embodiment, immunoconjugates with more than one agent may be prepared in a variety of ways. For example, more than one agent may be coupled directly to an antibody molecule. Alternatively, linkers that provide multiple sites for attachment can be used.
[0136] As is well known in the art, a given polypeptide or polynucleotide may vary in its immunogenicity. It is often necessary therefore to couple the immunogen (e.g., a polypeptide or polynucleotide) of the present invention with a carrier. Exemplary and preferred carriers are CRM197, E coli (LT) toxin, V. cholera (CT) toxin, keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA). Other albumins such as ovalbumin, mouse serum albumin or rabbit serum albumin can also be used as carriers.
[0137] Where a CVHPP (or a fragment thereof) and a carrier protein are conjugated (i.e., covalently associated), conjugation may be any chemical method, process or genetic technique commonly used in the art. For example, a CVHPP (or a fragment thereof) and a carrier protein, may be conjugated by techniques, including, but not limited to: (1) direct coupling via protein functional groups (e.g., thiol-thiol linkage, amine-carboxyl linkage, amine-aldehyde linkage; enzyme direct coupling); (2) homobifunctional coupling of amines (e.g., using bis-aldehydes); (3) homobifunctional coupling of thiols (e.g., using bis-maleimides); (4) homobifunctional coupling via photoactivated reagents (5) heterobifunctional coupling of amines to thiols (e.g., using maleimides); (6) heterobifunctional coupling via photoactivated reagents (e.g., the β-carbonyldiazo family); (7) introducing amine-reactive groups into a poly- or oligosaccharide via cyanogen bromide activation or carboxymethylation; (8) introducing thiol-reactive groups into a poly- or oligosaccharide via a heterobifunctional compound such as maleimido-hydrazide; (9) protein-lipid conjugation via introducing a hydrophobic group into the protein and (10) protein-lipid conjugation via incorporating a reactive group into the lipid. Also, contemplated are heterobifunctional "non-covalent coupling" techniques such the Biotin-Avidin interaction. For a comprehensive review of conjugation techniques, see Aslam and Dent (Aslam and Dent, "Bioconjugation: Protein Coupling Techniques for the Biomedical Sciences," Macmillan Reference Ltd., London, England, 1998), incorporated hereinafter by reference in its entirety.
[0138] In a specific embodiment, antibodies to a CVHPP may be used to eliminate the CVHPP in vivo by activating the complement system or mediating antibody-dependent cellular cytotoxicity (ADCC), or cause uptake of the antibody coated cells by the receptor-mediated endocytosis (RE) system.
Vectors
[0139] Another aspect of the invention pertains to vectors containing a polynucleotide encoding a CVHPP, a variant of a CVHPP, or a portion thereof. One type of vector is a "plasmid," which includes a circular double-stranded DNA loop into which additional DNA segments can be ligated. In the present specification, "plasmid" and "vector" can be used interchangeably as the plasmid is the most commonly used form of vector. Vectors also include expression vectors and gene delivery vectors.
[0140] The expression vectors of the invention comprise a polynucleotide encoding a CVHPP or a portion thereof in a form suitable for expression of the polynucleotide in a host cell, which means that the expression vectors include one or more regulatory sequences, selected on the basis of the host cells to be used for expression, and operatively linked to the polynucleotide sequence to be expressed. It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, and the like. The expression vectors of the invention can be introduced into host cells to thereby produce proteins or peptides, such as CVHPPs, mutant forms of CVHPPs, CVHPP-fusion proteins, and the like.
[0141] The expression vectors of the invention can be designed for expression of CVHPPs in prokaryotic or eukaryotic cells. For example, CVHPPs can be expressed in bacterial cells such as E. coli, insect cells (using baculovirus expression vectors), yeast cells or mammalian cells. Alternatively, the expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.
[0142] The expression of proteins in prokaryotes is most often carried out in E. coli with vectors containing constitutive or inducible promoters directing the expression of either fusion or non-fusion proteins. Fusion vectors add a number of amino acids to a protein encoded therein, usually to the amino terminus of the recombinant protein. Such fusion vectors typically serve three purposes: 1) to increase expression of the recombinant protein; 2) to increase the solubility of the recombinant protein; and 3) to aid in the purification of the recombinant protein by acting as a ligand in affinity purification. Often, in fusion expression vectors, a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant protein to enable separation of the recombinant protein from the fusion moiety subsequent to purification of the fusion protein. Such enzymes, and their cognate recognition sequences, include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia, Piscataway, N.J.), pMAL (New England Biolabs, Beverly, Mass.) and pRITS (Pharmacia, Piscataway, N.J.) which fuse glutathione S transferase (GST), maltose E binding protein, and protein A, respectively, to the target recombinant protein.
[0143] Purified fusion proteins can be utilized in CVHPP activity assays, (e.g., direct assays or competitive assays described in detail below), or to generate antibodies specific for CVHPPs.
[0144] One strategy to maximize recombinant protein expression in E. coli is to express the protein in a host bacteria with an impaired capacity to proteolytically cleave the recombinant protein. Another strategy is to alter the polynucleotide sequence of the polynucleotide to be inserted into an expression vector so that the individual codons for each amino acid are those preferentially utilized in E. coli. Such alteration of polynucleotide sequences of the invention can be carried out by standard DNA synthesis techniques.
[0145] In another embodiment, the CVHPP expression vector is a yeast expression vector. Examples of vectors for expression in yeast S. cerevisiae include pYepSec1, pMFa, pJRY88, pYES2 and picZ (Invitrogen Corp, San Diego, Calif.).
[0146] Alternatively, CVHPPs of the invention can be expressed in insect cells using baculovirus expression vectors. Baculovirus vectors available for expression of proteins in cultured insect cells (e.g., Sf9 cells) include the pAc series and the pVL series.
[0147] In yet another embodiment, a polynucleotide of the invention is expressed in mammalian cells using a mammalian expression vector. Examples of mammalian expression vectors include pCDM8 and pMT2PC. When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, adenovirus 2 and 5, cytomegalovirus and Simian Virus 40. Target gene expression from the pTrc vector relies on host RNA polymerase transcription from a hybrid trp-lac fusion promoter. Target gene expression from the pET 11d vector relies on transcription from a T7 gn10-lac fusion promoter mediated by a coexpressed viral RNA polymerase (T7 gn1). This viral polymerase is supplied by host strains BL21 (DE3) or HSLE174(DE3) from a resident prophage harboring a T7 gn1 gene under the transcriptional control of the lacUV 5 promoter.
[0148] In another embodiment, the mammalian expression vector is capable of directing expression of the polynucleotide preferentially in a particular cell type (e.g., tissue-specific regulatory elements are used to express the polynucleotide). Tissue-specific regulatory elements are known in the art and may include epithelial cell-specific promoters. Other non-limiting examples of suitable tissue-specific promoters include the liver-specific promoter (e.g., albumin promoter), lymphoid-specific promoters, promoters of T cell receptors and immunoglobulins, neuron-specific promoters (e.g., the neurofilament promoter), pancreas-specific promoters (e.g., insulin promoter), and mammary gland-specific promoters (e.g., milk whey promoter). Developmentally-regulated promoters (e.g., the α-fetoprotein promoter) are also encompassed.
[0149] The invention provides a recombinant expression vector comprising a polynucleotide encoding a CVHPP cloned into the expression vector in an antisense orientation. That is, the DNA molecule is operatively linked to a regulatory sequence in a manner which allows for expression (by transcription of the DNA molecule) of an RNA molecule which is antisense to mRNA corresponding to a CVHG of the invention. Regulatory sequences operatively linked to a polynucleotide cloned in the antisense orientation can be chosen which direct the continuous expression of the antisense RNA molecule in a variety of cell types, for instance, viral promoters and/or enhancers, or regulatory sequences can be chosen which direct constitutive, tissue specific or cell type specific expression of antisense RNA. The antisense expression vector can be in the form of a recombinant plasmid, phagemid or attenuated virus in which antisense polynucleotides are produced under the control of a high efficiency regulatory region, the activity of which can be determined by the cell type into which the vector is introduced.
[0150] The invention further provides gene delivery vehicles for delivery of polynucleotides to cells, tissues, or a mammal for expression. For example, a polynucleotide sequence of the invention can be administered either locally or systemically in a gene delivery vehicle. These constructs can utilize viral or non-viral vector approaches in in vivo or ex vivo modality. Expression of the coding sequence can be induced using endogenous mammalian or heterologous promoters. Expression of the coding sequence in vivo can be either constituted or regulated. The invention includes gene delivery vehicles capable of expressing the contemplated polynucleotides. The gene delivery vehicle is preferably a viral vector and, more preferably, a retroviral, lentiviral, adenoviral, adeno-associated viral (AAV), herpes viral, or alphavirus vector. The viral vector can also be an astrovirus, coronavirus, orthomyxovirus, papovavirus, paramyxovirus, parvovirus, picornavirus, poxvirus, togavirus viral vector.
[0151] The delivery of gene therapy constructs of this invention into cells is not limited to the above mentioned viral vectors. Other delivery methods and media may be employed such as, for example, nucleic acid expression vectors, polycationic condensed DNA linked or unlinked to killed adenovirus alone, ligand linked DNA, liposomes, eukaryotic cell delivery vehicles cells, deposition of photopolymerized hydrogel materials, handheld gene transfer particle gun, ionizing radiation, nucleic charge neutralization or fusion with cell membranes. Particle mediated gene transfer may be employed. Briefly, DNA sequence can be inserted into conventional vectors that contain conventional control sequences for high level expression, and then be incubated with synthetic gene transfer molecules such as polymeric DNA-binding cations like polylysine, protamine, and albumin, linked to cell targeting ligands such as asialoorosomucoid, insulin, galactose, lactose or transferrin. Naked DNA may also be employed. Uptake efficiency of naked DNA may be improved using biodegradable latex beads. The method may be improved further by treatment of the beads to increase hydrophobicity and thereby facilitate disruption of the endosome and release of the DNA into the cytoplasm.
[0152] Another aspect of the invention pertains to the expression of CVHPPs using a regulatable expression system. Systems to regulate expression of therapeutic genes have been developed and incorporated into the current viral and nonviral gene delivery vectors. These systems are briefly described below:
[0153] Tet-on/off system. The Tet-system is based on two regulatory elements derived from the tetracycline-resistance operon of the E. coli Tn10 transposon: the tet repressor protein (TetR) and the Tet operator DNA sequence (tetO) to which TetR binds. The system consists of two components, a "regulator" and a "reporter" plasmid. The "regulator" plasmid encodes a hybrid protein containing a mutated Tet repressor (rtetR) fused to the VP16 activation domain of herpes simplex virus. The "reporter" plasmid contains a tet-responsive element (TRE), which controls the "reporter" gene of choice. The rtetR-VP16 fusion protein can only bind to the TRE, therefore activates the transcription of the "reporter" gene, in the presence of tetracycline. The system has been incorporated into a number of viral vectors including retrovirus, adenovirus and AAV.
[0154] Ecdysone system. The ecdysone system is based on the molting induction system found in Drosophila, but modified for inducible expression in mammalian cells. The system uses an analog of the drosophila steroid hormone ecdysone, muristerone A, to activate expression of the gene of interest via a heterodimeric nuclear receptor. Expression levels have been reported to exceed 200-fold over basal levels with no effect on mammalian cell physiology.
[0155] Progesterone system. The progesterone receptor is normally stimulated to bind to a specific DNA sequence and to activate transcription through an interaction with its hormone ligand. Conversely, the progesterone antagonist mifepristone (RU486) is able to block hormone-induced nuclear transport and subsequent DNA binding. A mutant form of the progesterone receptor that can be stimulated to bind through an interaction with RU486 has been generated. To generate a specific, regulatable transcription factor, the RU486-binding domain of the progesterone receptor has been fused to the DNA-binding domain of the yeast transcription factor GAL4 and the transactivation domain of the HSV protein VP16. The chimeric factor is inactive in the absence of RU486. The addition of hormone, however, induces a conformational change in the chimeric protein, and this change allows binding to a GAL4-binding site and the activation of transcription from promoters containing the GAL4-binding site.
[0156] Rapamycin system. Immunosuppressive agents, such as FK506 and rapamycin, act by binding to specific cellular proteins and facilitating their dimerization. For example, the binding of rapamycin to FK506-binding protein (FKBP) results in its heterodimerization with another rapamycin binding protein FRAP, which can be reversed by removal of the drug. The ability to bring two proteins together by addition of a drug potentiates the regulation of a number of biological processes, including transcription. A chimeric DNA-binding domain has been fused to the FKBP, which enables binding of the fusion protein to a specific DNA-binding sequence. A transcriptional activation domain also has been fused to FRAP. When these two fusion proteins are co-expressed in the same cell, a fully functional transcription factor can be formed by heterodimerization mediated by addition of rapamycin. The dimerized chimeric transcription factor can then bind to a synthetic promoter sequence containing copies of the synthetic DNA-binding sequence. This system has been successfully integrated into adenoviral and AAV vectors. Long term regulatable gene expression has been achieved in both mice and baboons.
Immunogens and Immunogenic Compositions
[0157] Within certain aspects, CVHPP, CVHPN, CVHPP-specific T cell, CVHPP-presenting APC, CVHG-containing vectors, including but are not limited to expression vectors and gene delivery vectors, may be utilized as vaccines for CVH. Vaccines may comprise one or more such compounds/cells and an immunostimulant. An immunostimulant may be any substance that enhances or potentiates an immune response (antibody and/or cell-mediated) to an exogenous antigen. Examples of immunostimulants include adjuvants, biodegradable microspheres (e.g., polylactic galactide) and liposomes (into which the compound is incorporated). Vaccines within the scope of the present invention may also contain other compounds, which may be biologically active or inactive. For example, one or more immunogenic portions of other antigens may be present, either incorporated into a fusion polypeptide or as a separate compound, within the composition of vaccine.
[0158] A vaccine may contain DNA encoding one or more CVHPP or portion of CVHPP, such that the polypeptide is generated in situ. As noted above, the DNA may be present within any of a variety of delivery systems known to those of ordinary skill in the art, including nucleic acid expression vectors, gene delivery vectors, and bacteria expression systems. Numerous gene delivery techniques are well known in the art. Appropriate nucleic acid expression systems contain the necessary DNA sequences for expression in the patient (such as a suitable promoter and terminating signal). Bacterial delivery systems involve the administration of a bacterium (such as Bacillus-Calmette-Guerrin) that expresses an immunogenic portion of the polypeptide on its cell surface or secretes such an epitope. In a preferred embodiment, the DNA may be introduced using a viral expression system (e.g., vaccinia or other pox virus, retrovirus, or adenovirus), which may involve the use of a non-pathogenic (defective), replication competent virus. Techniques for incorporating DNA into such expression systems are well known to those of ordinary skill in the art. The DNA may also be "naked," as described, for example, in Ulmer et al., Science 259:1745-1749, 1993 and reviewed by Cohen, Science 259:1691-1692, 1993. The uptake of naked DNA may be increased by coating the DNA onto biodegradable beads, which are efficiently transported into the cells. It will be apparent that a vaccine may comprise both a polynucleotide and a polypeptide component. Such vaccines may provide for an enhanced immune response.
[0159] It will be apparent that a vaccine may contain pharmaceutically acceptable salts of the polynucleotides and polypeptides provided herein. Such salts may be prepared from pharmaceutically acceptable non-toxic bases, including organic bases (e.g., salts of primary, secondary and tertiary amines and basic amino acids) and inorganic bases (e.g., sodium, potassium, lithium, ammonium, calcium and magnesium salts).
[0160] Any of a variety of immunostimulants may be employed in the vaccines of this invention. For example, an adjuvant may be included. As defined previously, an "adjuvant" is a substance that serves to enhance the immunogenicity of an antigen. Thus, adjuvants are often given to boost the immune response and are well known to the skilled artisan. Examples of adjuvants contemplated in the present invention include, but are not limited to, aluminum salts (alum) such as aluminum phosphate and aluminum hydroxide, Mycobacterium tuberculosis, Bordetella pertussis, bacterial lipopolysaccharides, aminoalkyl glucosamine phosphate compounds (AGP), or derivatives or analogs thereof, which are available from Corixa (Hamilton, Mont.), and which are described in U.S. Pat. No. 6,113,918; one such AGP is 2-[(R)-3-Tetradecanoyloxytetradecanoylamino]ethyl 2-Deoxy-4-O-phosphono-3-O--[(R)-3-tetradecanoyoxytetradecanoyl]-2-[(R)-3-- tetradecanoyoxytetradecanoylamino]-b-D-glucopyranoside, which is also known as 529 (formerly known as RC529), which is formulated as an aqueous form or as a stable emulsion, MPL® (3-O-deacylated monophosphoryl lipid A) (Corixa) described in U.S. Pat. No. 4,912,094, synthetic polynucleotides such as oligonucleotides containing a CpG motif (U.S. Pat. No. 6,207,646), polypeptides, saponins such as Quil A or STIMULON® QS-21 (Antigenics, Framingham, Mass.), described in U.S. Pat. No. 5,057,540, a pertussis toxin (PT), or an E. coli heat-labile toxin (LT), particularly LT-K63, LT-R72, CT-S109, PT-K9/G129; see, e.g., International Patent Publication Nos. WO 93/13302 and WO 92/19265, cholera toxin (either in a wild-type or mutant form, e.g., wherein the glutamic acid at amino acid position 29 is replaced by another amino acid, preferably a histidine, in accordance with published International Patent Application number WO 00/18434). Various cytokines and lymphokines are suitable for use as adjuvants. One such adjuvant is granulocyte-macrophage colony stimulating factor (GM-CSF), which has a nucleotide sequence as described in U.S. Pat. No. 5,078,996. A plasmid containing GM-CSF cDNA has been transformed into E. coli and has been deposited with the American Type Culture Collection (ATCC), 1081 University Boulevard, Manassas, Va. 20110-2209, under Accession Number 39900. The cytokine IL-12 is another adjuvant which is described in U.S. Pat. No. 5,723,127. Other cytokines or lymphokines have been shown to have immune modulating activity, including, but not limited to, the interleukins 1-alpha, 1-beta, 2, 4, 5, 6, 7, 8, 10, 13, 14, 15, 16, 17 and 18, the interferons-alpha, beta and gamma, granulocyte colony stimulating factor, and the tumor necrosis factors alpha and beta, and are suitable for use as adjuvants.
[0161] Any vaccine provided herein may be prepared using well known methods that result in a combination of antigen, immune response enhancer and a suitable carrier or excipient. The compositions described herein may be administered as part of a sustained release formulation (i.e., a formulation such as a capsule, sponge or gel (composed of polysaccharides, for example) that effects a slow release of compound following administration). Such formulations may generally be prepared using well known technology and administered by, for example, oral, rectal or subcutaneous implantation, or by implantation at the desired target site. Sustained-release formulations may contain a polypeptide, polynucleotide or antibody dispersed in a carrier matrix and/or contained within a reservoir surrounded by a rate controlling membrane.
[0162] Carriers for use within such formulations are biocompatible, and may also be biodegradable; preferably the formulation provides a relatively constant level of active component release. Such carriers include microparticles of poly(lactide-co-glycolide), as well as polyacrylate, latex, starch, cellulose and dextran. Other delayed-release carriers include supramolecular biovectors, which comprise a non-liquid hydrophilic core (e.g., a cross-linked polysaccharide or oligosaccharide) and, optionally, an external layer comprising an amphiphilic compound, such as a phospholipid (see e.g., U.S. Pat. No. 5,151,254 and PCT applications WO 94/20078, WO/94/23701 and WO 96/06638). The amount of active compound contained within a sustained release formulation depends upon the site of implantation, the rate and expected duration of release and the nature of the condition to be treated or prevented.
[0163] Any of a variety of delivery vehicles may be employed within vaccines to facilitate production of an antigen-specific immune response that targets cancer cells. Delivery vehicles include antigen presenting cells (APCs), such as dendritic cells, macrophages, B cells, monocytes and other cells that may be engineered to be efficient APCs. Such cells may, but need not, be genetically modified to increase the capacity for presenting the antigen, to improve activation and/or maintenance of the T cell response, to have anti-CVH effects per se and/or to be immunologically compatible with the receiver (i.e., matched HLA haplotype). APCs may generally be isolated from any of a variety of biological fluids and organs, and may be autologous, allogeneic, syngeneic or xenogenic cells.
[0164] Certain preferred embodiments of the present invention use dendritic cells or progenitors thereof as APCs. Dendritic cells are highly potent APCs and have been shown to be effective as a physiological adjuvant for eliciting prophylactic or therapeutic anti-CVH immunity. In general, dendritic cells may be identified based on their typical shape (stellate in situ, with marked cytoplasmic processes (dendrites) visible in vitro), their ability to take up, process and present antigens with high efficiency and their ability to activate naive T cell responses. Dendritic cells may, of course, be engineered to express specific cell-surface receptors or ligands that are not commonly found on dendritic cells in vivo, and such modified dendritic cells are contemplated by the present invention. As an alternative to dendritic cells, secreted vesicles antigen-loaded dendritic cells (called exosomes) may be used within a vaccine (see Zitvogel et al., Nature Med. 4:594-600, 1998).
[0165] Dendritic cells and progenitors may be obtained from peripheral blood, bone marrow, lymph nodes, spleen, skin, umbilical cord blood or any other suitable tissue or fluid. For example, dendritic cells may be differentiated ex vivo by adding a combination of cytokines such as GM-CSF, IL-4, IL-13 and/or TNFα to cultures of monocytes harvested from peripheral blood. Alternatively, CD34 positive cells harvested from peripheral blood, umbilical cord blood or bone marrow may be differentiated into dendritic cells by adding to the culture medium combinations of GM-CSF, IL-3, TNFα, CD40 ligand, LPS, flt3 ligand and/or other compound(s) that induce differentiation, maturation and proliferation of dendritic cells.
[0166] Dendritic cells are conveniently categorized as "immature" and "mature" cells, which allows a simple way to discriminate between two well characterized phenotypes. However, this nomenclature should not be construed to exclude all possible intermediate stages of differentiation. Immature dendritic cells are characterized as APC with a high capacity for antigen uptake and processing, which correlates with the high expression of Fcy receptor and mannose receptor. The mature phenotype is typically characterized by a lower expression of these markers, but a high expression of cell surface molecules responsible for T cell activation such as class I and class II MHC, adhesion molecules (e.g., CD54 and CD11) and costimulatory molecules (e.g., CD40, CD80, CD86 and 4-1BB).
[0167] APCs may generally be transfected with a polynucleotide encoding a CVHPP (or portion or other variant thereof) such that the CVHPP, or an immunogenic portion thereof, is expressed on the cell surface. Such transfection may take place ex vivo, and a composition or vaccine comprising such transfected cells may then be used for therapeutic purposes, as described herein. Alternatively, a gene delivery vehicle that targets a dendritic or other antigen presenting cell may be administered to a patient, resulting in transfection that occurs in vivo. In vivo and ex vivo transfection of dendritic cells, for example, may generally be performed using any methods known in the art, such as those described in WO 97/24447, or the gene gun approach described by Mahvi et al., Immunology and cell Biology 75:456-460, 1997. Antigen loading of dendritic cells may be achieved by incubating dendritic cells or progenitor cells with the CVHPP, DNA (naked or within a plasmid vector) or RNA; or with antigen-expressing recombinant bacterium or viruses (e.g., vaccinia, fowlpox, adenovirus or lentivirus vectors). Prior to loading, the polypeptide may be covalently conjugated to an immunological partner that provides T cell help (e.g., a carrier molecule). Alternatively, a dendritic cell may be pulsed with a non-conjugated immunological partner, separately or in the presence of the polypeptide.
[0168] Vaccines may be presented in unit-dose or multi-dose containers, such as sealed ampoules or vials. Such containers are preferably hermetically sealed to preserve sterility of the formulation until use. In general, formulations may be stored as suspensions, solutions or emulsions in oily or aqueous vehicles. Alternatively, a vaccine may be stored in a freeze-dried condition requiring only the addition of a sterile liquid carrier immediately prior to use.
Detection Methods
[0169] As discussed earlier, expression level of CVHGs may be used as a marker for CVH and CVH-related disorders. Detection and measurement of the relative amount of a CVHG product (polynucleotide or polypeptide) of the invention can be by any method known in the art. Typical methodologies for detection of a transcribed polynucleotide include RNA extraction from a cell or tissue sample, followed by hybridization of a labeled probe (i.e., a complementary polynucleotide molecule) specific for the target RNA to the extracted RNA and detection of the probe (i.e., Northern blotting).
[0170] Typical methodologies for peptide detection include protein extraction from a cell or tissue sample, followed by binding of an antibody specific for the target protein to the protein sample, and detection of the antibody. For example, detection of desmin may be accomplished using polyclonal antibody anti-desmin. Antibodies are generally detected by the use of a labeled secondary antibody. The label can be a radioisotope, a fluorescent compound, an enzyme, an enzyme co-factor, or ligand. Such methods are well understood in the art.
[0171] In certain embodiments, the CVHGs themselves (i.e., the DNA or cDNA) may serve as markers for CVH. For example, an increase of genomic copies of a CVHG, such as by duplication of the gene, may also be correlated with CVH and CVH-associated disorders.
[0172] Detection of specific polynucleotide molecules may also be assessed by gel electrophoresis, column chromatography, or direct sequencing, quantitative PCR (in the case of polynucleotide molecules), RT-PCR, or nested-PCR among many other techniques well known to those skilled in the art.
[0173] Detection of the presence or number of copies of all or a part of an CVHG of the invention may be performed using any method known in the art. Typically, it is convenient to assess the presence and/or quantity of a DNA or cDNA by Southern analysis, in which total DNA from a cell or tissue sample is extracted and hybridized with a labeled probe (i.e., a complementary DNA molecules). The probe is then detected and quantified. The label group can be a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor. Other useful methods of DNA detection and/or quantification include direct sequencing, gel electrophoresis, column chromatography, and quantitative PCR, as is known by one skilled in the art.
[0174] In certain embodiments, CVHPPs may serve as markers for CVH. Detection of specific polypeptide molecules may be assessed by gel electrophoresis, Western blot, column chromatography, or direct sequencing, among many other techniques well known to those skilled in the art.
Panels of CVHGs
[0175] The expression level of each CVHG may be considered individually, although it is within the scope of the invention to provide combinations of two or more CVHGs for use in the methods and compositions of the invention to increase the confidence of the analysis. In another aspect, the invention provides panels of the CVHGs of the invention. A panel of CVHGs comprises two or more CVHGs. A panel may also comprise 2-5, 5-15, 15-35, 35-50, or more than 50 CVHGs. In a preferred embodiment, these panels of CVHGs are selected such that the CVHGs within any one panel share certain features. For example, the CVHGs of a first panel may all relate to myelination in a CVH sample. Alternatively, CVHGs of a second panel may each exhibit differential regulation as compared to a first panel. Similarly, different panels of CVHGs may be composed of CVHGs representing different stages of CVH. Panels of the CVHGs of the invention may be made by independently selecting CVHGs from Tables 3-8, and may further be provided on biochips, as discussed below.
Screening Methods
[0176] The invention also provides methods (also referred to herein as "screening assays") for identifying modulators, i.e., candidate or test compounds or agents comprising therapeutic moieties (e.g., peptides, peptidomimetics, peptoids, polynucleotides, small molecules or other drugs) which (a) bind to a CVHPP, or (b) have a modulatory (e.g., stimulatory or inhibitory) effect on the activity of a CVHPP or, more specifically, (c) have a modulatory effect on the interactions of the CVHPP with one or more of its natural substrates (e.g., peptide, protein, hormone, co-factor, or polynucleotide), or (d) have a modulatory effect on the expression of the CVHPPs. Such assays typically comprise a reaction between the CVHPP and one or more assay components. The other components may be either the test compound itself, or a combination of the test compound and a binding partner of the CVHPP.
[0177] To screen for compounds which interfere with binding of two proteins e.g., a CVHPP and its binding partner, a Scintillation Proximity Assay can used. In this assay, the CVHPP is labeled with an isotope such as 125I. The binding partner is labeled with a scintillant, which emits light when proximal to radioactive decay (i.e., when the CVHPP is bound to its binding partner). A reduction in light emission will indicate that a compound has interfered with the binding of the two proteins.
[0178] Alternatively a Fluorescence Energy Transfer (FRET) assay could be used. In a FRET assay of the invention, a fluorescence energy donor is comprised on one protein (e.g., a CVHPP) and a fluorescence energy acceptor is comprised on a second protein (e.g., a binding partner of the CVHPP). If the absorption spectrum of the acceptor molecule overlaps with the emission spectrum of the donor fluorophore, the fluorescent light emitted by the donor is absorbed by the acceptor. The donor molecule can be a fluorescent residue on the protein (e.g., intrinsic fluorescence such as a tryptophan or tyrosine residue), or a fluorophore which is covalently conjugated to the protein (e.g., fluorescein isothiocyanate, FITC). An appropriate donor molecule is then selected with the above acceptor/donor spectral requirements in mind.
[0179] Thus, in this example, a CVHPP is labeled with a fluorescent molecule (i.e., a donor fluorophore) and its binding partner is labeled with a quenching molecule (i.e., an acceptor). When the CVHPP and its binding partner are bound, fluorescence emission will be quenched or reduced relative the CVHPP alone. Similarly, a compound which can dissociate the interaction of the CVHPP-partner complex, will result in an increase in fluorescence emission, which indicates the compound has interfered with the binding of the CVHPP to its binding partner.
[0180] Another assay to detect binding or dissociation of two proteins is fluorescence polarization or anisotropy. In this assay, the investigated protein (e.g., a CVHPP) is labeled with a fluorophore with an appropriate fluorescence lifetime. The protein sample is then excited with vertically polarized light. The value of anisotropy is then calculated by determining the intensity of the horizontally and vertically polarized emission light. Next, the labeled protein (CVHPP) is mixed with a CVHPP binding partner and the anisotropy measured again. Because fluorescence anisotropy intensity is related to the rotational freedom of the labeled protein, the more rapidly a protein rotates in solution, the smaller the anisotropy value. Thus, if the labeled CVHPP is part of a complex (e.g., CVHPP-partner), the CVHPP rotates more slowly in solution (relative to free, unbound CVHPP) and the anisotropy intensity increases. Subsequently, a compound which can dissociate the interaction of the CVHPP-partner complex, will result in a decrease in anisotropy (i.e., the labeled CVHPP rotates more rapidly), which indicates the compound has interfered with the binding of CVHPP to its binding partner.
[0181] A more traditional assay would involve labeling the CVHPP binding partner with an isotope such as 125I, incubating with the CVHPP, then immunoprecipitating of the CVHPP. Compounds that increase the free CVHPP will decrease the precipitated counts. To avoid using radioactivity, the CVHPP binding partner could be labeled with an enzyme-conjugated antibody instead.
[0182] Alternatively, the CVHPP binding partner could be immobilized on the surface of an assay plate and the CVHPP could be labeled with a radioactive tag. A rise in the number of counts would identify compounds that had interfered with binding of the CVHPP and its binding partner.
[0183] Evaluation of binding interactions may further be performed using Biacore technology, wherein the CVHPP or its binding partner is bound to a micro chip, either directly by chemical modification or tethered via antibody-epitope association (e.g., antibody to the CVHPP), antibody directed to an epitope tag (e.g., His tagged) or fusion protein (e.g., GST). A second protein or proteins is/are then applied via flow over the "chip" and the change in signal is detected. Finally, test compounds are applied via flow over the "chip" and the change in signal is detected.
[0184] Once a series of potential compounds has been identified for a combination of CVHPP, CVHPP binding partner and ALS, a bioassay can be used to select the most promising candidates. For example, a cellular assay that measures cell proliferation in presence of the CVHPP and the CVHPP binding partner was described above. This assay could be modified to test the effectiveness of small molecules that interfere with binding of a CVHPP and its binding partner in enhancing cellular proliferation. An increase in cell proliferation would correlate with a compound's potency.
[0185] The test compounds of the present invention are generally either small molecules or biomolecules. Small molecules include, but are not limited to, inorganic molecules and small organic molecules. Biomolecules include, but are not limited to, naturally-occurring and synthetic compounds that have a bioactivity in mammals, such as lipids, steroids, polypeptides, polysaccharides, and polynucleotides. In one preferred embodiment, the test compound is a small molecule. In another preferred embodiment, the test compound is a biomolecule. One skilled in the art will appreciate that the nature of the test compound may vary depending on the nature of the CVHPP. For example, if the CVHPP is an orphan receptor having an unknown ligand, the test compound may be any of a number of biomolecules which may act as cognate ligand, including but not limited to, cytokines, lipid-derived mediators, small biogenic amines, hormones, neuropeptides, or proteases.
[0186] The test compounds of the present invention may be obtained from any available source, including systematic libraries of natural and/or synthetic compounds. Test compounds may also be obtained by any of the numerous approaches in combinatorial library methods known in the art, including: biological libraries; peptoid libraries (libraries of molecules having the functionalities of peptides, but with a novel, non-peptide backbone which are resistant to enzymatic degradation but which nevertheless remain bioactive); spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; the `one-bead one-compound` library method; and synthetic library methods using affinity chromatography selection. The biological library and peptoid library approaches are limited to peptide libraries, while the other four approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds. As used herein, the term "binding partner" refers to a molecule which serves as either a substrate for a CVHPP, or alternatively, as a ligand having binding affinity to the CVHPP.
Screening for Inhibitors of CVHPP
[0187] The invention provides methods of screening test compounds for inhibitors of CVHPP. The method of screening comprises obtaining samples from subjects diagnosed with or suspected of having CVH, contacting each separate aliquot of the samples with one of a plurality of test compounds, and comparing expression of one or more CVHGs in each of the aliquots to determine whether any of the test compounds provides a substantially altered level of expression or activity of an CVHG relative to samples with other test compounds or relative to an untreated sample or control sample. In addition, methods of screening may be devised by combining a test compound with a protein and thereby determining the effect of the test compound on the protein.
[0188] In addition, the invention is further directed to a method of screening for test compounds capable of modulating with the binding of a CVHPP and a binding partner, by combining the test compound, CVHPP, and binding partner together and determining whether binding of the binding partner and CVHPP occurs. The test compound may be either small molecules or a bioactive agent. As discussed below, test compounds may be provided from a variety of libraries well known in the art.
[0189] Modulators of CVHG expression, activity or binding ability are useful as therapeutic compositions of the invention. Such modulators (e.g., antagonists or agonists) may be formulated as pharmaceutical compositions, as described herein below. Such modulators may also be used in the methods of the invention, for example, to diagnose, treat, or prognose CVH.
High-Throughput Screening Assays
[0190] The invention provides methods of conducting high-throughput screening for test compounds capable of inhibiting activity or expression of a CVHPP of the present invention. In one embodiment, the method of high-throughput screening involves combining test compounds and the CVHPP and detecting the effect of the test compound on the CVHPP.
[0191] A variety of high-throughput functional assays well-known in the art may be used in combination to screen and/or study the reactivity of different types of activating test compounds. Since the coupling system is often difficult to predict, a number of assays may need to be configured to detect a wide range of coupling mechanisms. A variety of fluorescence-based techniques are well-known in the art and are capable of high-throughput and ultra high throughput screening for activity, including but not limited to BRET® or FRET® (both by Packard Instrument Co., Meriden, Conn.). The ability to screen a large volume and a variety of test compounds with great sensitivity permits analysis of the therapeutic targets of the invention to further provide potential inhibitors of CVH. For example, where the CVHG encodes an orphan receptor with an unidentified ligand, high-throughput assays may be utilized to identify the ligand, and to further identify test compounds which prevent binding of the receptor to the ligand. The BIACORE® system may also be manipulated to detect binding of test compounds with individual components of the therapeutic target, to detect binding to either the encoded protein or to the ligand.
[0192] By combining test compounds with CVHPPs of the invention and determining the binding activity between them, diagnostic analysis can be performed to elucidate the coupling systems. Generic assays using cytosensor microphysiometer may also be used to measure metabolic activation, while changes in calcium mobilization can be detected by using the fluorescence-based techniques such as FLIPR® (Molecular Devices Corp, Sunnyvale, Calif.). In addition, the presence of apoptotic cells may be determined by TUNEL assay, which utilizes flow cytometry to detect free 3-OH termini resulting from cleavage of genomic DNA during apoptosis. As mentioned above, a variety of functional assays well-known in the art may be used in combination to screen and/or study the reactivity of different types of activating test compounds. Preferably, the high-throughput screening assay of the present invention utilizes label-free plasmon resonance technology as provided by BIACORE® systems (Biacore International AB, Uppsala, Sweden). Plasmon free resonance occurs when surface plasmon waves are excited at a metal/liquid interface. By reflecting directed light from the surface as a result of contact with a sample, the surface plasmon resonance causes a change in the refractive index at the surface layer. The refractive index change for a given change of mass concentration at the surface layer is similar for many bioactive agents (including proteins, peptides, lipids and polynucleotides), and since the BIACORE® sensor surface can be functionalized to bind a variety of these bioactive agents, detection of a wide selection of test compounds can thus be accomplished.
[0193] Therefore, the invention provides for high-throughput screening of test compounds for the ability to inhibit activity of a protein encoded by the CVHGs listed in Table 4, by combining the test compounds and the protein in high-throughput assays such as BIACORE®, or in fluorescence-based assays such as BRET®. In addition, high-throughput assays may be utilized to identify specific factors which bind to the encoded proteins, or alternatively, to identify test compounds which prevent binding of the receptor to the binding partner. In the case of orphan receptors, the binding partner may be the natural ligand for the receptor. Moreover, the high-throughput screening assays may be modified to determine whether test compounds can bind to either the encoded protein or to the binding partner (e.g., substrate or ligand) which binds to the protein.
Predictive Medicine
[0194] The present invention pertains to the field of predictive medicine in which diagnostic assays, prognostic assays, pharmacogenetics and monitoring clinical trials are used for prognostic (predictive) purpose to thereby treat an individual prophylactically. Accordingly, one aspect of the present invention relates to diagnostic assays for determining CVHG expression and/or activity, in the context of a biological sample (e.g., blood, urine, feces, serum, cells, tissue) to thereby determine whether an individual is at risk for developing CVH associated with altered CVHG expression or activity. The invention also provides for prognostic (or predictive) assays for determining whether an individual is at risk of developing CVH associated with aberrant CVHG expression or activity.
[0195] For example, the number of copies of an CVHG can be assayed in a biological sample. Such assays can be used for prognostic or predictive purposes to thereby prophylactically treat an individual prior to the onset of CVH associated with aberrant CVHG protein, polynucleotide expression or activity.
[0196] Another aspect of the invention pertains to monitoring the influence of agents (e.g., drugs, compounds) on the expression or activity of CVHGs in clinical trials.
Diagnostic Assays
[0197] An exemplary method for detecting the presence or absence of a CVHPP or CVHPN in a biological sample involves contacting a biological sample with a compound or an agent capable of detecting the CVHPP or CVHPN (e.g., mRNA, genomic DNA). A preferred agent for detecting mRNA or genomic DNA corresponding to an CVHG or CVHPP of the invention is a labeled polynucleotide probe capable of hybridizing to a mRNA or genomic DNA of the invention. In a most preferred embodiment, the polynucleotides to be screened are arranged on a GeneChip®. Suitable probes for use in the diagnostic assays of the invention are described herein. A preferred agent for detecting a CVHPP of the invention is an antibody which specifically recognizes the CVHPP.
[0198] The diagnostic assays may also be used to quantify the amount of expression or activity of a CVHG in a biological sample. Such quantification is useful, for example, to determine the progression or severity of CVH and CVH-related disorders. Such quantification is also useful, for example, to determine the severity of CVH following treatment.
Determining Severity of CVH and CVH-Related Diseases
[0199] In the field of diagnostic assays, the invention also provides methods for determining the severity of CVH by isolating a sample from a subject (e.g., a colon biopsy), and detecting the presence, quantity and/or activity of one or more CVHG products in the sample relative to a second sample from a normal sample or control sample. In one embodiment, the expression levels of CVHGs in the two samples are compared, and a modulation in one or more CVHGs in the test sample indicates CVH. In other embodiments the modulation of 2, 3, 4 or more CVHGs indicates a severe case of CVH.
[0200] In another aspect, the invention provides CVHG products whose quantity or activity is correlated with the severity of CVH. The subsequent level of expression may further be compared to different expression profiles of various stages of the disease to confirm whether the subject has a matching profile. In yet another aspect, the invention provides CVHGs whose quantity or activity is correlated with a risk in a subject for developing CVH.
[0201] A preferred agent for detecting a CVHPP is an antibody capable of binding to the CVHPP, preferably an antibody with a detectable label. Antibodies can be polyclonal or more preferably, monoclonal. An intact antibody, or a fragment thereof (e.g., Fab or F(ab')2) can be used. The term "labeled," with regard to the probe or antibody, is intended to encompass direct labeling of the probe or antibody by coupling (i.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling of the probe or antibody by reactivity with another reagent that is directly labeled. Examples of indirect labeling include detection of a primary antibody using a fluorescently labeled secondary antibody and end-labeling of a DNA probe with biotin such that it can be detected with fluorescently labeled streptavidin. The term "biological sample" is intended to include tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. That is, the detection method of the invention can be used to detect CVHG mRNA, protein or genomic DNA in a biological sample in vitro as well as in vivo. For example, in vitro techniques for detection of CVHG mRNA include Northern hybridizations and in situ hybridizations. In vitro techniques for detection of CVHPP include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations and immunofluorescence. In vitro techniques for detection of CVHG genomic DNA include Southern hybridizations. Furthermore, in vivo techniques for detection of CVHPP include introducing into a subject a labeled anti-CVHPP antibody. For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.
[0202] In one embodiment, the biological sample contains protein molecules from the test subject. Alternatively, the biological sample can contain mRNA molecules from the test subject or genomic DNA molecules from the test subject. A preferred biological sample is a tissue or serum sample isolated by conventional means from a subject, e.g., a biopsy or blood draw.
Prognostic Assays
[0203] The diagnostic method described herein can be utilized to identify subjects having or at risk of developing CVH associated with aberrant CVHG expression or activity.
[0204] The assays described herein, such as the preceding or following assays, can be utilized to identify a subject having CVH associated with an aberrant level of CVHG activity or expression. Alternatively, the prognostic assays can be utilized to identify a subject at risk for developing CVH associated with aberrant levels of CVHG protein activity or polynucleotide expression. Thus, the present invention provides a method for identifying CVH associated with aberrant CVHG expression or activity in which a test sample is obtained from a subject and CVHPP or CVHPN (e.g., mRNA or genomic DNA) is detected, wherein the presence of CVHPP or CVHPN is diagnostic or prognostic for a subject having or at risk of developing CVH with aberrant CVHG expression or activity.
[0205] Furthermore, the prognostic assays described herein can be used to determine whether a subject can be administered an agent (e.g., an agonist, antagonist, peptidomimetic, protein, peptide, polynucleotide, small molecule, or other drug candidate) to treat or prevent CVH associated with aberrant CVHG expression or activity, such as, for example, a cytokine. For example, such methods can be used to determine whether a subject can be effectively treated with an agent to inhibit CVH. Thus, the present invention provides methods for determining whether a subject can be effectively treated with an agent for CVH associated with aberrant CVHG expression or activity.
[0206] Prognostic assays can be devised to determine whether a subject undergoing treatment for CVH has a poor outlook for disease progression. In a preferred embodiment, prognosis can be determined shortly after diagnosis, i.e., within a few days. By establishing expression profiles of different stages of CVHGs, from onset to later stages, an expression pattern may emerge to correlate a particular expression profile to increased likelihood of a poor prognosis. The prognosis may then be used to devise a more aggressive treatment program and enhance the likelihood of success.
[0207] The methods of the invention can also be used to detect genetic alterations in a CVHG, thereby determining if a subject with the altered gene is at risk for damage characterized by aberrant regulation in CVHG expression or activity. In preferred embodiments, the methods include detecting, in a sample of cells from the subject, the presence or absence of a genetic alteration characterized by at least one alteration affecting the integrity of a CVHG, or the aberrant expression of the CVHG. For example, such genetic alterations can be detected by ascertaining the existence of at least one of the following: 1) deletion of one or more nucleotides from a CVHG; 2) addition of one or more nucleotides to a CVHG; 3) substitution of one or more nucleotides of a CVHG, 4) a chromosomal rearrangement of a CVHG; 5) alteration in the level of a messenger RNA transcript of a CVHG, 6) aberrant modification of a CVHG, such as of the methylation pattern of the genomic DNA, 7) the presence of a non-wild type splicing pattern of a messenger RNA transcript of a CVHG, 8) non-wild type level of a CVHPP, 9) allelic loss of a CVHG, and 10) inappropriate post-translational modification of a CVHPP. As described herein, there are a large number of assays known in the art, which can be used for detecting alterations in a CVHG or a CVHG product. A preferred biological sample is a blood sample isolated by conventional means from a subject.
[0208] In certain embodiments, detection of the alteration involves the use of a probe/primer in a polymerase chain reaction (PCR), such as anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR), the latter of which can be particularly useful for detecting point mutations in the CVHG. This method can include the steps of collecting a sample of cells from a subject, isolating a polynucleotide sample (e.g., genomic, mRNA or both) from the cells of the sample, contacting the polynucleotide sample with one or more primers which specifically hybridize to a CVHG under conditions such that hybridization and amplification of the CVHG (if present) occurs, and detecting the presence or absence of an amplification product, or detecting the size of the amplification product and comparing the length to a control sample. It is understood that PCR and/or LCR may be desirable to be used as a preliminary amplification step in conjunction with any of the techniques used for detecting mutations described herein.
[0209] Alternative amplification methods include: self-sustained sequence replication, transcriptional amplification system, Q-Beta Replicase, or any other polynucleotide amplification method, followed by the detection of the amplified molecules using techniques well known to those of skill in the art. These detection schemes are especially useful for the detection of polynucleotide molecules if such molecules are present in very low numbers.
[0210] In an alternative embodiment, mutations in an CVHG from a sample cell can be identified by alterations in restriction enzyme cleavage patterns. For example, sample and control DNA is isolated, amplified (optionally), digested with one or more restriction endonucleases, and fragment length sizes are determined by gel electrophoresis and compared. Differences in fragment length sizes between sample and control DNA indicate mutations in the sample DNA. Moreover, sequence specific ribozymes (see, for example, U.S. Pat. No. 5,498,531) can be used to score for the presence of specific mutations by development or loss of a ribozyme cleavage site.
[0211] In other embodiments, genetic mutations in a CVHG can be identified by hybridizing sample and control polynucleotides, e.g., DNA or RNA, to high density arrays containing hundreds or thousands of oligonucleotides probes. For example, genetic mutations in a CVHG can be identified in two dimensional arrays containing light generated DNA probes. Briefly, a first hybridization array of probes can be used to scan through long stretches of DNA in a sample and control to identify base changes between the sequences by making linear arrays of sequential overlapping probes. This step allows the identification of point mutations. This step is followed by a second hybridization array that allows the characterization of specific mutations by using smaller, specialized probe arrays complementary to all variants or mutations detected. Each mutation array is composed of parallel probe sets, one complementary to the wild-type gene and the other complementary to the mutant gene.
[0212] In yet another embodiment, any of a variety of sequencing reactions known in the art can be used to directly sequence the CVHG and detect mutations by comparing the sequence of the sample CVHG with the corresponding wild-type (control) sequence. It is also contemplated that any of a variety of automated sequencing procedures can be utilized when performing the diagnostic assays, including sequencing by mass spectrometry.
[0213] Other methods for detecting mutations in a CVHG include methods in which protection from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA/DNA heteroduplexes. In general, the art technique of "mismatch cleavage" starts by providing heteroduplexes by hybridizing (labeled) RNA or DNA containing the wild-type CVHG sequence with potentially mutant RNA or DNA obtained from a tissue sample. The double-stranded duplexes are treated with an agent which cleaves single-stranded regions of the duplex, which will exist due to basepair mismatches between the control and sample strands. For instance, RNA/DNA duplexes can be treated with RNase and DNA/DNA hybrids treated with S1 nuclease to enzymatically digest the mismatched regions. In other embodiments, either DNA/DNA or RNA/DNA duplexes can be treated with hydroxylamine or osmium tetroxide and with piperidine in order to digest mismatched regions. After digestion of the mismatched regions, the resulting material is then separated by size on denaturing polyacrylamide gels to determine the site of mutation. In a preferred embodiment, the control DNA or RNA can be labeled for detection.
[0214] In still another embodiment, the mismatch cleavage reaction employs one or more proteins that recognize mismatched base pairs in double-stranded DNA (so called "DNA mismatch repair" enzymes) in defined systems for detecting and mapping point mutations in CVHG cDNAs obtained from samples of cells. For example, the mutY enzyme of E. coli cleaves A at G/A mismatches and the thymidine DNA glycosylase from HeLa cells cleaves T at G/T mismatches. According to an exemplary embodiment, a probe based on an CVHG sequence, e.g., a wild-type CVHG sequence, is hybridized to cDNA or other DNA product from a test cell(s). The duplex is treated with a DNA mismatch repair enzyme, and the cleavage products, if any, can be detected from electrophoresis protocols or the like. See, for example, U.S. Pat. No. 5,459,039.
[0215] In other embodiments, alterations in electrophoretic mobility will be used to identify mutations in CVHGs. For example, single-strand conformation polymorphism (SSCP) may be used to detect differences in electrophoretic mobility between mutant and wild type polynucleotides. Single-stranded DNA fragments of sample and control CVHG polynucleotides will be denatured and allowed to renature. The secondary structure of single-stranded polynucleotides varies according to sequence. The resulting alteration in electrophoretic mobility enables the detection of even a single base change. The DNA fragments may be labeled or detected with labeled probes. The sensitivity of the assay may be enhanced by using RNA (rather than DNA) in which the secondary structure is more sensitive to a change in sequence. In a preferred embodiment, the subject method utilizes heteroduplex analysis to separate double-stranded heteroduplex molecules on the basis of changes in electrophoretic mobility (Keen et al. Trends Genet. 7:5, 1991).
[0216] In yet another embodiment the movement of mutant or wild-type fragments in polyacrylamide gels containing a gradient of denaturant is assayed using denaturing gradient gel electrophoresis (DGGE). When DGGE is used as the method of analysis, DNA will be modified to insure that it does not completely denature, for example, by adding a GC clamp of approximately 40 bp of high-melting GC-rich DNA by PCR. In a further embodiment, a temperature gradient is used in place of a denaturing gradient to identify differences in the mobility of control and sample DNA (Rosenbaum and Reissner Biophys Chem 265:12753, 1987).
[0217] Examples of other techniques for detecting point mutations include, but are not limited to, selective oligonucleotide hybridization, selective amplification, and selective primer extension. For example, oligonucleotide primers may be prepared in which the known mutation is placed centrally and then hybridized to target DNA under conditions which permit hybridization only if a perfect match is found (Saiki et al. Proc. Natl. Acad. Sci. USA 86:6230, 1989). Such allele specific oligonucleotides are hybridized to PCR amplified target or a number of different mutations when the oligonucleotides are attached to the hybridizing membrane and hybridized with labeled target DNA.
[0218] Alternatively, allele specific amplification technology which depends on selective PCR amplification may be used in conjunction with the instant invention. Oligonucleotides used as primers for specific amplification may carry the mutation of interest in the center of the molecule (so that amplification depends on differential hybridization) or at the extreme 3' end of one primer where, under appropriate conditions, mismatch can prevent or reduce polymerase extension. In addition, it may be desirable to introduce a novel restriction site in the region of the mutation to create cleavage-based detection. It is anticipated that, in certain embodiments, amplification may also be performed using Taq ligase for amplification. In such cases, ligation will occur only if there is a perfect match at the 3' end of the 5' sequence, thus making it possible to detect the presence of a known mutation at a specific site by looking for the presence or absence of amplification.
[0219] The methods described herein may be performed, for example, by utilizing prepackaged diagnostic kits comprising at least one probe polynucleotide or antibody reagent described herein, which may be conveniently used, e.g., in clinical settings to diagnose subjects exhibiting symptoms or family history of a disease or illness involving a CVHG.
[0220] Furthermore, any cell type or tissue in which a CVHG is expressed may be utilized in the prognostic or diagnostic assays described herein.
Monitoring Effects During Clinical Trials
[0221] Monitoring the influence of agents (e.g., drugs, small molecules, proteins, nucleotides) on the expression or activity of a CVHPP can be applied not only in basic drug screening, but also in clinical trials. For example, the effectiveness of an agent determined by a screening assay, as described herein to decrease CVHG expression or activity, can be monitored in clinical trials of subjects exhibiting increased CVHG expression or activity. In such clinical trials, the expression or activity of an CVHG can be used as a "read-out" of the phenotype of a particular tissue.
[0222] For example, and not by way of limitation, CVHGs that are modulated in tissues by treatment with an agent can be identified. Thus, to study the effect of agents on the CVHPP in a clinical trial, cells can be isolated and RNA prepared and analyzed for the levels of expression of an CVHG. The levels of gene expression or a gene expression pattern can be quantified by Northern blot analysis, RT-PCR or GeneChip® as described herein, or alternatively by measuring the amount of protein produced, by one of the methods as described herein, or by measuring the levels of activity of CVHPP. In this way, the gene expression pattern can serve as a read-out, indicative of the physiological response of the cells to the agent. Accordingly, this response state may be determined before treatment and at various points during treatment of the individual with the agent.
[0223] In a preferred embodiment, the present invention provides a method for monitoring the effectiveness of treatment of a subject with an agent (e.g., an agonist, antagonist, peptidomimetic, protein, peptide, polynucleotide, small molecule, or other drug candidate identified by the screening assays described herein) including the steps of (i) obtaining a pre-administration sample from a subject prior to administration of the agent; (ii) detecting the level of expression of a CVHG protein or mRNA in the pre-administration sample; (iii) obtaining one or more post-administration samples from the subject; (iv) detecting the level of expression or activity of the CVHG protein or mRNA in the post-administration samples; (v) comparing the level of expression or activity of the CVHG protein or mRNA in the pre-administration sample with the CVHG protein or mRNA the post administration sample or samples; and (vi) altering the administration of the agent to the subject accordingly. According to such an embodiment, CVHG expression or activity may be used as an indicator of the effectiveness of an agent, even in the absence of an observable phenotypic response.
Methods of Treatment
[0224] The present invention provides for both prophylactic and therapeutic methods of treating a subject at risk for, susceptible to or diagnosed with CVH. With regard to both prophylactic and therapeutic methods of treatment, such treatments may be specifically tailored or modified, based on knowledge obtained from the field of pharmacogenomics. "Pharmacogenomics," as used herein, includes the application of genomics technologies such as gene sequencing, statistical genetics, and gene expression analysis to drugs in clinical development and on the market. More specifically, the term refers the study of how a subject's genes determine his or her response to a drug (e.g., a subject's "drug response phenotype" or "drug response genotype"). Thus, another aspect of the invention provides methods for tailoring an individual's prophylactic or therapeutic treatment with either the CVHPP molecules of the present invention or CVHPP modulators (e.g., agonists or antagonists) according to that individual's drug response. Pharmacogenomics allows a clinician or physician to target prophylactic or therapeutic treatments to subjects who will most benefit from the treatment and to avoid treatment of subjects who will experience toxic drug-related side effects.
Prophylactic Methods
[0225] In one aspect, the invention provides a method for preventing in a subject CVH associated with aberrant CVHG expression or activity, by administering to the subject anCVHG product or an agent which modulates CVHG protein expression or activity.
[0226] Subjects at risk for CVH which is caused or contributed to by aberrant CVHG expression or activity can be identified by, for example, any or a combination of diagnostic or prognostic assays as described herein.
[0227] Administration of a prophylactic agent can occur prior to the manifestation of symptoms characteristic of the differential CVHG protein expression, such that CVH is prevented or, alternatively, delayed in its progression. Depending on the type of CVHG aberrancy (e.g., typically a modulation outside the normal standard deviation), for example, a CVHG product, CVHG agonist or antagonist agent can be used for treating the subject. The appropriate agent can be determined based on screening assays described herein.
Therapeutic Methods
[0228] Another aspect of the invention pertains to methods of modulating CVHG protein expression or activity for therapeutic purposes. Accordingly, in an exemplary embodiment, the modulatory method of the invention involves contacting a cell with an agent that modulates one or more of the activities of a CVHG product activity associated with the cell. An agent that modulates CVHG product activity can be an agent as described herein, such as a polynucleotide (e.g., an antisense molecule) or a polypeptide (e.g., a dominant-negative mutant of a CVHPP), a naturally-occurring target molecule of a CVHPP (e.g., a CVHPP substrate), an anti-CVHPP antibody, a CVHG modulator (e.g., agonist or antagonist), a peptidomimetic of a CVHG protein agonist or antagonist, or other small molecules.
[0229] The invention further provides methods of modulating a level of expression of a CVHG of the invention, comprising administration to a subject having CVH, a variety of compositions which correspond to the CVHGs of Tables 3-8, including proteins or antisense oligonucleotides. The protein may be provided by further providing a vector comprising a polynucleotide encoding the protein to the cells. Alternatively, the expression levels of the CVHGs of the invention may be modulated by providing an antibody, a plurality of antibodies or an antibody conjugated to a therapeutic moiety. Treatment with the antibody may further be localized to the tissue comprising CVH. In another aspect, the invention provides methods for localizing a therapeutic moiety to CVH tissue or cells comprising exposing the tissue or cells to an antibody which is specific to a protein encoded by the CVHGs of the invention. This method may therefore provide a means to inhibit expression of a specific gene corresponding to a CVHG listed in Tables 3-8.
Determining Efficacy of a Test Compound or Therapy
[0230] The invention also provides methods of assessing the efficacy of a test compound or therapy for inhibiting CVH in a subject. These methods involve isolating samples from a subject suffering from CVH, who is undergoing treatment or therapy, and detecting the presence, quantity, and/or activity of one or more CVHGs of the invention in the first sample relative to a second sample. Where the efficacy of a test compound is determined, the first and second samples are preferably sub-portions of a single sample taken from the subject, wherein the first portion is exposed to the test compound and the second portion is not. In one aspect of this embodiment, the CVHG is expressed at a substantially decreased level in the first sample, relative to the second. Most preferably, the level of expression in the first sample approximates (i.e., is less than the standard deviation for normal samples) the level of expression in a third control sample, taken from a control sample of normal tissue. This result suggests that the test compound inhibits the expression of the CVHG in the sample. In another aspect of this embodiment, the CVHG is expressed at a substantially increased level in the first sample, relative to the second. Most preferably, the level of expression in the first sample approximates (i.e., is less than the standard deviation for normal samples) the level of expression in a third control sample, taken from a control sample of normal tissue. This result suggests that the test compound augments the expression of the CVHG in the sample.
[0231] Where the efficacy of a therapy is being assessed, the first sample obtained from the subject is preferably obtained prior to provision of at least a portion of the therapy, whereas the second sample is obtained following provision of the portion of the therapy. The levels of CVHG product in the samples are compared, preferably against a third control sample as well, and correlated with the presence, or risk of presence, of CVH. Most preferably, the level of CVHG product in the second sample approximates the level of expression of a third control sample. In the present invention, a substantially decreased level of expression of a CVHG indicates that the therapy is efficacious for treating CVH.
Pharmacogenomics
[0232] The CVHG protein and polynucleotide molecules of the present invention, as well as agents, inhibitors or modulators which have a stimulatory or inhibitory effect on CVHG expression or activity as identified by a screening assay described herein, can be administered to individuals to treat (prophylactically or therapeutically) CVH associated with aberrant CVHG activity.
[0233] In conjunction with such treatment, pharmacogenomics (i.e., the study of the relationship between an individual's genotype and that individual's response to a foreign compound or drug) may be considered. Differences in metabolism of therapeutics can lead to severe toxicity or therapeutic failure by altering the relation between dose and blood concentration of the pharmacologically active drug. Thus, a physician or clinician may consider applying knowledge obtained in relevant pharmacogenomics studies in determining whether to administer a CVHG product (polynucleotide or polypeptide) or CVHG modulator as well as tailoring the dosage and/or therapeutic regimen of treatment with a CVHG product or CVHG modulator.
[0234] Pharmacogenomics deals with clinically significant hereditary variations in the response to drugs due to altered drug disposition and abnormal action in affected persons. In general, two types of pharmacogenetic conditions can be differentiated. Genetic conditions transmitted as a single factor altering the way drugs act on the body (altered drug action) or genetic conditions transmitted as single factors altering the way the body acts on drugs (altered drug metabolism). These pharmacogenetic conditions can occur either as rare genetic defects or as naturally-occurring polymorphisms. For example, glucose-6-phosphate dehydrogenase deficiency (G6PD) is a common inherited enzymopathy in which the main clinical complication is hemolysis after ingestion of oxidant drugs (anti-malarials, sulfonamides, analgesics, nitrofurans) and consumption of fava beans.
[0235] One pharmacogenomics approach to identifying genes that predict drug response, known as "a genome-wide association," relies primarily on a high-resolution map of the human genome consisting of already known gene-related sites (e.g., a "bi-allelic" gene marker map which consists of 60,000-100,000 polymorphic or variable sites on the human genome, each of which has two variants). Such a high-resolution genetic map can be compared to a map of the genome of each of a statistically substantial number of subjects taking part in a Phase II/III drug trial to identify genes associated with a particular observed drug response or side effect. Alternatively, such a high resolution map can be generated from a combination of some ten-million known single nucleotide polymorphisms (SNPs) in the human genome. As used herein, an "SNP" is a common alteration that occurs in a single nucleotide base in a stretch of DNA. For example, an SNP may occur once per every 1,000 bases of DNA. An SNP may be involved in a disease process. However, the vast majority of SNPs may not be disease associated. Given a genetic map based on the occurrence of such SNPs, individuals can be grouped into genetic categories depending on a particular pattern of SNPs in their individual genome. In such a manner, treatment regimens can be tailored to groups of genetically similar individuals, taking into account traits that may be common among such genetically similar individuals. Thus, mapping of the CVHGs of the invention to SNP maps of CVH patients may allow easier identification of these genes according to the genetic methods described herein.
[0236] Alternatively, a method termed the "candidate gene approach," can be utilized to identify genes that predict drug response. According to this method, if a gene that encodes a drug target is known (e.g., a CVHG of the present invention), all common variants of that gene can be fairly easily identified in the population and it can be determined if having one version of the gene versus another is associated with a particular drug response.
[0237] As an illustrative embodiment, the activity of drug metabolizing enzymes is a major determinant of both the intensity and duration of drug action. The discovery of genetic polymorphisms of drug metabolizing enzymes (e.g., N-acetyltransferase 2 (NAT 2) and cytochrome P450 enzymes CYP2D6 and CYPZC19) has provided an explanation as to why some subjects do not obtain the expected drug effects or show exaggerated drug response and serious toxicity after taking the standard and safe dose of a drug. These polymorphisms are expressed in two phenotypes in the population, the extensive metabolizer and poor metabolizer. The prevalence of poor metabolizer phenotypes is different among different populations. For example, the gene coding for CYP2D6 is highly polymorphic and several mutations have been identified in poor metabolizers, which all lead to the absence of functional CYP2D6. Poor metabolizers of CYP2D6 and CYP2C19 quite frequently experience exaggerated drug response and side effects when they receive standard doses. If a metabolite is the active therapeutic moiety, poor metabolizers show no therapeutic response, as demonstrated for the analgesic effect of codeine mediated by its CYP2D6-formed metabolite morphine. The other extreme are the so called ultra-rapid metabolizers who do not respond to standard doses. Recently, the molecular basis of ultra-rapid metabolism has been identified to be due to CYP2D6 gene amplification.
[0238] Alternatively, a method termed the "gene expression profiling" can be utilized to identify genes that predict drug response. For example, the gene expression of an animal dosed with a drug (e.g., CVHG expression in response to a CVHG modulator of the present invention) can give an indication whether gene pathways related to toxicity have been turned on.
[0239] Information generated from more than one of the above pharmacogenomics approaches can be used to determine appropriate dosage and treatment regimens for prophylactic or therapeutic treatment an individual. This knowledge, when applied to dosing or drug selection, can avoid adverse reactions or therapeutic failure and thus enhance therapeutic or prophylactic efficiency when treating a subject with a CVHG product or CVHG modulator, such as a modulator identified by one of the exemplary screening assays described herein.
Pharmaceutical Compositions
[0240] The invention is further directed to pharmaceutical compositions comprising the test compound, or bioactive agent, or an CVHG modulator (i.e., agonist or antagonist), which may further include a CVHG product, and can be formulated as described herein. Alternatively, these compositions may include an antibody which specifically binds to a CVHG protein of the invention and/or an antisense polynucleotide molecule which is complementary to a CVHG polynucleotide of the invention and can be formulated as described herein.
[0241] One or more of the CVHGs of the invention, fragments of CVHGs, CVHG products, fragments of CVHG products, CVHG modulators, or anti-CVHPP antibodies of the invention can be incorporated into pharmaceutical compositions suitable for administration.
[0242] As used herein the language "pharmaceutically acceptable carrier" is intended to include any and all solvents, solubilizers, fillers, stabilizers, binders, absorbents, bases, buffering agents, lubricants, controlled release vehicles, diluents, emulsifying agents, humectants, lubricants, dispersion media, coatings, antibacterial or antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well-known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary agents can also be incorporated into the compositions.
[0243] The invention includes methods for preparing pharmaceutical compositions for modulating the expression or activity of a polypeptide or polynucleotide corresponding to a CVHG of the invention. Such methods comprise formulating a pharmaceutically acceptable carrier with an agent which modulates expression or activity of a CVHG. Such compositions can further include additional active agents. Thus, the invention further includes methods for preparing a pharmaceutical composition by formulating a pharmaceutically acceptable carrier with an agent which modulates expression or activity of a CVHG and one or more additional bioactive agents.
[0244] A pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (topical), intraperitoneal, transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine; propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfate; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates or phosphates and agents for the adjustment of tonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.
[0245] Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL® (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the injectable composition should be sterile and should be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the requited particle size in the case of dispersion and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.
[0246] Sterile injectable solutions can be prepared by incorporating the active compound (e.g., a fragment of a CVHPP or an anti-CVHPP antibody) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
[0247] Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part of the composition. The tablets, pills, capsules, troches and the like can contain any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose; a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate or Stertes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring.
[0248] For administration by inhalation, the compounds are delivered in the form of an aerosol spray from a pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.
[0249] Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the bioactive compounds are formulated into ointments, salves, gels, or creams as generally known in the art.
[0250] The compounds can also be prepared in the form of suppositories (e.g., with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.
[0251] In one embodiment, the therapeutic moieties, which may contain a bioactive compound, are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from e.g. Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art.
[0252] It is especially advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form, as used herein, includes physically discrete units suited as unitary dosages for the subject to be treated; each unit contains a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the invention are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.
[0253] Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED50. Compounds which exhibit large therapeutic indices are preferred. While compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets such compounds to the site of affected tissue in order to minimize potential damage to uninfected cells and, thereby, reduce side effects.
[0254] The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of such compounds lies preferably within a range of circulating concentrations that includes the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.
[0255] The CVHGs of the invention can be inserted into gene delivery vectors and used as gene therapy vectors. Gene therapy vectors can be delivered to a subject by, for example, intravenous administration, intraportal administration, intrabiliary administration, intra-arterial administration, direct injection into the liver parenchyma, by intramuscular injection, by inhalation, by perfusion, or by stereotactic injection. The pharmaceutical preparation of the gene therapy vector can include the gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. Alternatively, where the complete gene delivery vector can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical preparation can include one or more cells which produce the gene delivery system.
[0256] The pharmaceutical compositions can be included in a container, pack, or dispenser together with instructions for administration.
Kits
[0257] The invention also encompasses kits for detecting the presence of a CVHG product in a biological sample, the kit comprising reagents for assessing expression of the CVHGs of the invention. Preferably, the reagents may be an antibody or fragment thereof, wherein the antibody or fragment thereof specifically binds with a protein corresponding to a CVHG from Table 4. For example, antibodies of interest may be prepared by methods known in the art. Optionally, the kits may comprise a polynucleotide probe wherein the probe specifically binds with a transcribed polynucleotide corresponding to a CVHG selected from the group consisting of the CVHGs listed in Tables 3-8. The kits may also include an array of CVHGs arranged on a biochip, such as, for example, a GeneChip®. The kit may contain means for determining the amount of the CVHG protein or mRNA in the sample; and means for comparing the amount of the CVHG protein or mRNA in the sample with a control or standard. The compound or agent can be packaged in a suitable container. The kit can further comprise instructions for using the kit to detect CVHG protein or polynucleotide
[0258] The invention further provides kits for assessing the suitability of each of a plurality of compounds for inhibiting CVH in a subject. Such kits include a plurality of compounds to be tested, and a reagent (i.e., antibody specific to corresponding proteins, or a probe or primer specific to corresponding polynucleotides) for assessing expression of a CVHG listed in Tables 3-8.
Arrays and Biochips
[0259] The invention also includes an array comprising a panel of CVHGs of the present invention. The array can be used to assay expression of one or more genes in the array.
[0260] It will be appreciated by one skilled in the art that the panels of CVHGs of the invention may conveniently be provided on solid supports, such as a biochip. For example, polynucleotides may be coupled to an array (e.g., a biochip using GeneChip® for hybridization analysis), to a resin (e.g., a resin which can be packed into a column for column chromatography), or a matrix (e.g., a nitrocellulose matrix for Northern blot analysis). The immobilization of molecules complementary to the CVHG(s), either covalently or noncovalently, permits a discrete analysis of the presence or activity of each CVHG in a sample. In an array, for example, polynucleotides complementary to each member of a panel of CVHGs may individually be attached to different, known locations on the array. The array may be hybridized with, for example, polynucleotides extracted from a blood or colon sample from a subject. The hybridization of polynucleotides from the sample with the array at any location on the array can be detected, and thus the presence or quantity of the CVHG and CVHG transcripts in the sample can be ascertained. In a preferred embodiment, an array based on a biochip is employed. Similarly, Western analyses may be performed on immobilized antibodies specific for CVHPPs hybridized to a protein sample from a subject.
[0261] It will also be apparent to one skilled in the art that the entire CVHG product (protein or polynucleotide) molecule need not be conjugated to the biochip support; a portion of the CVHG product or sufficient length for detection purposes (i.e., for hybridization), for example a portion of the CVHG product which is 7, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 100 or more nucleotides or amino acids in length may be sufficient for detection purposes.
[0262] In one embodiment, the array can be used to assay gene expression in a tissue to ascertain tissue specificity of genes in the array. In this manner, a large number of genes can be simultaneously assayed for expression. This allows an expression profile to be developed showing a battery of genes specifically expressed in one or more tissues at a given point in time. In one embodiment the invention provides a kit comprising a brochure which comprises at least 5, more preferably 10, more preferably or more CVHGs, and the same CVHGs in computer readable form.
[0263] In addition to such qualitative determination, the invention allows the quantitation of gene expression in the biochip. Thus, not only tissue specificity, but also the level of expression of a battery of CVHGs in the tissue is ascertainable. Thus, CVHGs can be grouped on the basis of their tissue expression per se and level of expression in that tissue. As used herein, a "normal level of expression" refers to the level of expression of a gene provided in a control sample, typically the control is taken from either a non-diseased animal or from a subject who has not suffered from CVH. The determination of normal levels of expression is useful, for example, in ascertaining the relationship of gene expression between or among tissues. Thus, one tissue or cell type can be perturbed and the effect on gene expression in a second tissue or cell type can be determined. In this context, the effect of one cell type on another cell type in response to a biological stimulus can be determined. Such a determination is useful, for example, to know the effect of cell-cell interaction at the level of gene expression. If an agent is administered therapeutically to treat one cell type but has an undesirable effect on another cell type, the invention provides an assay to determine the molecular basis of the undesirable effect and thus provides the opportunity to co-administer a counteracting agent or otherwise treat the undesired effect. Similarly, even within a single cell type, undesirable biological effects can be determined at the molecular level. Thus, the effects of an agent on expression of other than the target gene can be ascertained and counteracted.
[0264] In another embodiment, the arrays can be used to monitor the time course of expression of one or more genes in the array. This can occur in various biological contexts, as disclosed herein, for example development and differentiation, disease progression, in vitro processes, such as cellular transformation and activation.
[0265] The array is also useful for ascertaining the effect of the expression of a gene on the expression of other genes in the same cell or in different cells. This provides, for example, for a selection of alternate molecular targets for therapeutic intervention if the ultimate or downstream target cannot be regulated.
[0266] Importantly, the invention provides arrays useful for ascertaining differential expression patterns of one or more genes identified in diseased tissue versus non-diseased tissue. This provides a battery of genes that serve as a molecular target for diagnosis or therapeutic intervention. In particular, biochips can be made comprising arrays not only of the CVHGs listed in Tables 3-8, but of CVHGs specific to subjects suffering from specific manifestations or stages of the disease.
[0267] In general, the probes are attached to the biochip in a wide variety of ways, as will be appreciated by those in the art. As described herein, the nucleic acids can either be synthesized first, with subsequent attachment to the biochip, or can be directly synthesized on the biochip.
[0268] The biochip comprises a suitable solid substrate. By "substrate" or "solid support" or other grammatical equivalents herein is meant any material that can be modified to contain discrete individual sites appropriate for the attachment or association of the nucleic acid probes and is amenable to at least one detection method. As will be appreciated by those in the art, the number of possible substrates are very large, and include, but are not limited to, glass and modified or functionalized glass, plastics (including acrylics, polystyrene and copolymers of styrene and other materials, polypropylene, polyethylene, polybutylene, polyurethanes, TeflonJ, etc.), polysaccharides, nylon or nitrocellulose, resins, silica or silica-based materials including silicon and modified silicon, carbon, metals, inorganic glasses, plastics, etc.
[0269] Generally the substrate is planar, although as will be appreciated by those in the art, other configurations of substrates may be used as well. For example, the probes may be placed on the inside surface of a tube, for flow-through sample analysis to minimize sample volume. Similarly, the substrate may be flexible, such as a flexible foam, including closed cell foams made of particular plastics.
[0270] In a preferred embodiment, the surface of the biochip and the probe may be derivatized with chemical functional groups for subsequent attachment of the two. Thus, for example, the biochip is derivatized with a chemical functional group including, but not limited to, amino groups, carboxy groups, oxo groups and thiol groups, with amino groups being particularly preferred. Using these functional groups, the probes can be attached using functional groups on the probes. For example, nucleic acids containing amino groups can be attached to surfaces comprising amino groups. Linkers, such as homo- or hetero-bifunctional linkers, may also be used.
[0271] In an embodiment, the oligonucleotides are synthesized as is known in the art, and then attached to the surface of the solid support. As will be appreciated by those skilled in the art, either the 5' or 3' terminus may be attached to the solid support, or attachment may be via an internal nucleoside.
[0272] In an additional embodiment, the immobilization to the solid support may be very strong, yet non-covalent. For example, biotinylated oligonucleotides can be made, which bind to surfaces covalently coated with streptavidin, resulting in attachment.
[0273] Alternatively, the oligonucleotides may be synthesized on the surface, as is known in the art. For example, photoactivation techniques utilizing photopolymerization compounds and techniques are used. In a preferred embodiment, the nucleic acids can be synthesized in situ, using well known photolithographic techniques.
[0274] Modifications to the above-described compositions and methods of the invention, according to standard techniques, will be readily apparent to one skilled in the art and are meant to be encompassed by the invention. This invention is further illustrated by the following examples which should not be construed as limiting. The contents of all references, patents and published patent applications cited throughout this application, as well as the Figures and Tables are incorporated herein by reference.
EXAMPLES
Example 1
Methods
(a) Visceral Sensitization Model
[0275] New born rats were sensitized by infusion of 0.2 mls of 0.5% acetic acid into the colon at P10, control animals received saline.
(b) CNI1493 Treatment:
[0276] At eight weeks of age, rats in four treatment groups: control+vehicle (n=3), control+cni-1493 (n=3), sensitized+vehicle (n=8), sensitized+cni-1493 (n=8) were tested after administration of drug (dissolved in 2.5% mannitol, injected i.p. at 5 mg/kg for four days) for sensitivity to colorectal distention (CRD). Vehicle injected animals received 2.5% mannitol in water.
(c) CRD Protocol for Measuring Colonic Sensitivity in Adult Rats
[0277] For electromyographic (EMG) measurements of visceromotor responses, under anesthesia (Nembutal, 50 mg/kg i.p.), two electrodes were implanted in the abdominal wall muscles and externalized behind the head. Rats were allowed a one week recovery time from this surgery. Under mild sedation (Brevital, 5 ml of 1% ip), a 7 cm flexible balloon constructed from a surgical glove finger attached to tygon tubing was inserted into the descending colon and rectum via the anus and held in place by taping the tubing to the tail. Rats were housed in small Lucite cubicles (20×8×8 cm) and were allowed to adapt for one hour. CRD was performed by rapidly inflating the balloon to constant pressure. Pressure was measured using a sphygmomanometer connected to a pressure transducer. Rats were given graded CRD (20, 40, 60, 80 mm Hg) applied for 20 seconds every 4 minutes.
[0278] The behavioral measurements consisted of visual observation of the abdominal withdrawal reflex (AWR) by blinded observers and the assignment of an AWR score:
[0279] 0, no response;
[0280] 1, brief head movement followed by immobility;
[0281] 2, contraction of abdominal muscles;
[0282] 3, lifting of abdomen;
[0283] 4, body arching and lifting of pelvic structures.
(d) Evaluation of the Colon for Inflammation/Damage
[0284] Descending colons were placed in 10% buffered formalin and portions were frozen for myeloperoxidase (MPO) assays. Haematoxilin & Eosin stained paraffin sections will be scored for inflammation by a pathologist.
(e) Tissue Isolation and RNA Preparation
[0285] Colons and S1 dorsal root ganglia were snap frozen in liquid nitrogen. Colon tissue RNA and S1 dorsal root ganglia RNA was prepared using RNAzol.
(f) Target Labeling and Gene Chip Array Analysis
[0286] An in vitro transcription reaction was performed using 500 ng cDNA which contained a T7 RNA polymerase promoter incorporated during RNA amplification. The cRNA or Target RNAs produced during this in vitro transcription reaction were labeled with biotin. Biotin-labeled Target RNAs were fragmented to a mean size of 200 bases to facilitate their hybridization to probe sequences on the Gene Chip (Affymetrix) array. Each Target RNA sample was initially hybridized to a test array. This array contains a set of probes representing genes that are commonly expressed in the majority of cells (example Rat: actin, GAPDH, hexokinase, 5S rRNA, and B1/B2 repetitive elements). Test arrays confirmed the successful labeling of the target RNAs and prevented the use of degraded or non-representative target RNA samples. Success and consistency of RNA amplification between samples were also confirmed with these test arrays.
[0287] Hybridization of Gene Chip (Affymetrix) arrays was performed at 45° C. for 16 hours in 0.1 M MES pH6.6, 1 M sodium chloride, 0.02 M EDTA and 0.01% Tween 20. Four prokaryotic genes (bio B, bio C and bio D from the E. coli biotin synthesis pathway and the cre recombinase gene from P1 bacteriophage) were added to the hybridization cocktail as internal controls. These control RNAs were used to normalize expression levels between experiments. Because the control RNAs were added at varying copy number (Bio B, 1.5 pM; Bio C, 5 pM; Bio D, 25 pM and cre, 100 pM) they were also used in estimating relative abundance of RNA transcripts in the sample. Arrays were washed using both non-stringent (1 M NaCl, 25° C.) and stringent (1 M NaCl, 50° C.) conditions prior to staining with phycoerythrin streptavidin (10 ug/ml final). Gene Chip arrays were scanned using a Gene Array Scanner (Hewlett Packard) and analyzed using the Affymetric Gene Chip Analysis Suite 5.0 software.
[0288] Gene expression profiles were produced from Affymetrix rat genome 230A chips.
[0289] Single array analysis for each chip was performed by Affymetrix Microarray Suite (MAS) software to produce a detection call, present, absent or marginal and a signal intensity value for each gene that is a relative measure of abundance of the transcript. For comparison of signal intensity values between chips, all chips were scaled to an average intensity of 500. Genes called "Absent" across all chips and genes without a |Fold change|≧2.0 in at least one of the pairwise comparisons of chips from different treatment groups were excluded. The probe sets with absolute call "Absent" across all chips and |Fold change|<2.0 in all of the possible pairwise comparisons were filtered out. These filters were applied as the first level filters to reduce the noise from the dataset. ANOVA was performed on the filtered dataset. Significant changes in gene expression were detected by analyzing signal intensity values by two-way ANOVA with 99% confidence limits. Genes were subjected to cluster analysis to identify genes associated with sensitization and treatment.
(g) Quantitative RT-PCR
[0290] Primer or primer/probe sets were designed using Primer Express software (Applied Biosystems, Foster City, Calif.) such that the amplimer spanned an intron/exon boundary where possible. Where ESTs were homologous to known genes, the sequence of the known gene was used. RT-PCR was performed using Applied Biosystems reagents and kits: Taqman Reverse Transcription Reagents N8080234 and Taqman PCR Core Reagents N8080228. PCR was performed on a GeneAmp 5700 Sequence Detection System (Applied Biosystems). Machine default PCR program was used: 2 min at 50° C., 10 min at 95° C., 45 cycles: 95° C. 15 sec, 60° C. 1 min. Fold change was calculated using delta Ct and/or relative standard curve procedures using GAPDH or b-actin as normalizers for colon genes and PGP9.5 for DRG. Data is expressed as fold change relative to control, vehicle treated values.
Primer Probe Sequences (RAT):
TABLE-US-00008 [0291] Desmin NM_022531 Forward primer (FP) GTGGAGCGTGACAACCTGATAG Reverse primer (RP) TGCGCTCTAGGTCAATTCGA CEBP/delta NM_013154 Mar. 3, 2004 SYBR green FP CCGCCCGAATTGCTACAGT RP AGTCTGTCGGAAAAGTCTTTTCTACAA EST homologous to F-box proteins FP CCGTGTGCAAGTGTGTAGCAT RP GCCGCAGCCCGAAAG PEP19 FP GCTGGAGCAACCAATGGAAA RP TCTGTCTCTGGTGCATCCATGT Probe TCT TCT TGG ACC TTC TT CTG CCC ATC ATT
Insulin-Like Growth Factor Binding Protein 2 Colon 71 NM--013122
TABLE-US-00009 [0292] FP 1001-1022 CAACCTCAAACAGTGCAAGATG RP 1182-1163 TGGTTTACTGCACCCTTTGG Metalloproteinase ADAMTS-1 FP AGGGACCGGAAGTTACTTCCA RP CAGGTGTGGGAGCCACATAA Tubulin, beta 3 FP GGGCCTTTGGACACCTATTCA RP GCCCTTTGGCCCAGTTGT PROBE C ACC ACT CTG ACC GAA GAT AAA GTT GTC AGG Argalb NM_053770 Feb. 19, 2004 FP GAATCCCCACAGCCATTAGAAC RP GCGAGTTGTACAGACCTGCATT Probe ACA TGT CTG TGT CCT CAT CCG GCT TGT Stathmin-like 2 (scgn10) FP TCGGAAGCTCCACGAACTCT RP CTCGCTCGTGCTCCCTCTT
(f) MPO Assay
[0293] Myeloperoxidase (MPO) assays were performed as described (Bhatia et al., Proc Natl Acad Sci USA 95:4760-4756, 1998). Frozen colon was homogenized in 20 mM phosphate buffer pH 7.4 and centrifuged at 10,000×g for 10 minutes at 4° C. Pellet was resuspended in 50 mM phosphate buffer, pH 6.0 containing 0.5% hexadecyltrimethylammonium bromide. Samples were subjected to one cycle of freeze thawing, were incubated at 60° C. for 2 hrs and centrifuged at 10,000×g for 5 min at 4° C. Change in absorbance at 655 nM was measured in reaction mix containing 1.6 mM TMB (Sigma) and 0.3 mM H2O2 on a Beckman DU-64 spectrophotometer. Activity was normalized to protein as measured in the extracts. Protein was measured by the BCA method (Pierce, Rockford, Ill.). Activity was expressed as the change in absorbance/min/mg protein.
Example 2
Chemical Treatment of Colon of P10 Rats Produces CVH
[0294] Previous studies showed that either mechanical irritation or treatment with mustard oil of the colons of young rats between P7 and P12 produced chronic visceral hyperalgesia (ref). To determine whether treatment of the colon of young rats with acetic acid would produce visceral hyperalgesia in adults, the colon of P10 rats was infused with 0.5% acetic acid; control littermates received saline. Rats were tested at 8 weeks of age for sensitivity to CRD and colons were examined for histopathological evidence of inflammation. Adult rats treated with acetic acid on P10 exhibited increased sensitivity to CRD compared to controls (FIG. 1). Data was analyzed by two-way repeated measures ANOVA with distention pressure as the repeated factor and P10 treatment as a between group factor. There was a significant effect of P10 treatment (F 1, 12.98) p<0.003, of distention pressure (F 7, 89.9) p<0.001, and there was a significant interaction between distention pressure and P10 treatment (F 7, 4.04) p<0.001. Means were compared with a Tukey test. Significant differences between AA treated and controls were found at distention pressures of 30 (p=0.004), 40 (p<0.001), 50 (p<0.001), 60 (p=0.001) and 70 (p=0.035) mm Hg. Chemical treatment of colon of P10 rats produces chronic visceral hypersensitivity.
Example 3
Effect of CNI1493 on Visceral Hypersensitivity
[0295] Rats were treated for 4 days with CNI1493 and sensitivity to CRD was measured after treatment. Sensitivity to CRD was significantly reduced by drug treatment F(1, 5.46) p=0.05 (FIG. 2) but was unchanged in vehicle treated rats. Data was analyzed by two-way repeated measures ANOVA with distention pressure as the repeated factor and drug treatment as a between group factor. There was a significant effect of CNI1493 treatment (F 1, 16.96) p=0.001, of distention pressure (F 7, 28.55) p<0.001, but there was no significant interaction between distention pressure and CNI-1493 treatment (F 7, 1.94) p=0.071. Means were compared with a Tukey test. Significant differences between CNI-1493 treated and controls were found at distention pressures of 20 (P=0.001), 30 (p=0.003), 40 (p<0.001), 50 (p=0.001), 60 (p<0.001) and 70 (p=0.015) and 80 (p=0.007) mm Hg.
[0296] To determine whether P10 colon irritation produced chronic inflammation, H&E stained colon sections were examined for signs of inflammation and MPO activity in colon extracts was examined. No differences in appearance of the sections were noted between treatment groups and no overt signs of inflammation were observed. Mucosal architecture was normal, there was no cellular infiltrate, and there was no depletion of goblet cells. No differences were observed in colon histopathology (FIG. 3A-3D). There was a significant increase in MPO activity in the sensitized rats compared to controls (FIG. 3E). CNI-1493 treatment significantly lowered MPO activity in sensitized rats. These findings suggest that there was a low grade inflammation or increased lymphocytes present in sensitized rats.
Example 4
Gene Expression Profiles in Colon and S1 DRG
[0297] To determine the long term effects of P10 colon irritation and of subsequent CNI1493 treatment on gene expression in the colon, gene expression profiles of rat colons from each treatment group derived from Affymetrix rat genome 230A chips were subjected to cluster analysis to identify genes associated with sensitization and treatment. The analysis revealed that 114 genes were differentially expressed in sensitized/CVH colon (Table 3), 76 genes were differentially expressed in sensitized/CVH S1 DRG (Table 4), 660 genes were differentially expressed in CNI1493-treated colon (Table 5), and 137 genes were differentially expressed in CNI1493-treated S1 DRG (Table 6). Since CNI1493 treatment ameliorates CVH in the sensitized animals, genes differentially regulated by CNI1493 may also be related to the etiology of CVH. Accordingly, genes listed in Tables 3-8 are designated as CVH-related genes (CVHGs). Expression levels of a subset of genes from each group were confirmed by quantitative RT-PCR (Table 8).
TABLE-US-00010 TABLE 8 Comparison of chip results with RT-PCR Gene sen + sen + sen + veh sen + CNI veh RT- CNI RT- Chip PCR Chip PCR desmin 2.6 1.7 1.3 0.8 CCAAT/enhancerbinding 2.4 2.2 2.3 1.6 (C/EBP) delta EST similar to F-box proteins 2.4 1.9 1.2 0.6 neuron specific protein PEP-19 2.3 2.1 1.1 1.1 Insulin-like growth factor binding 2.2 2.6 0.9 0.7 protein 2 ADAMTS1 2.1 2.0 1.1 0.8 tubulin, beta 3 2.1 2.9 1.5 1.3 ArgBP2 1.7 2.9 0.8 1.8 scgn10 1.5 1.3 1.1 0.4 BDNF 1.7 1.9 1.7 2.1 phosphodiesterase 3B 1.8 1.6 1.1 1.2 Trek2 2.1 1.5 1.7 0.9 TrkA precursor 0.7 0.8 0.7 0.7 interleukin 1 receptor, type 1 0.5 1.1 0.6 0.8 elongation factor 2 kinase 0.4 1.6 0.8 1.5
Sequence CWU
1
6612236DNAHomo sapiens 1cctcgccgca tccactctcc ggccggccgc ctgcccgccg
cctcctccgt gcgcccgcca 60gcctcgcccg cgccgtcacc atgagccagg cctactcgtc
cagccagcgc gtgtcctcct 120accgccgcac cttcggcggc gccccggtct tcccgctcgg
ctccccgctg agctcgcccg 180tgttcccgcg ggcgcctttc ggctctaagg gctcctccag
ctcggtgacg tcccgcgtgt 240accaggtgtc gcgcacgtcg ggaggggccg ggggcctggg
gtcgctgcgg gccagccggc 300tggggaccac ccgcacgccc tcctcctacg gcgcaggcga
gctgctggac ttctcactgg 360ccgacgcggt gaaccaggag tttctgacca cgcgcaccaa
cgagaaggtg gagctgcagg 420agctcaatga ccgttcgcca atctacatgg agaaggtgcg
cttcctggag cagcagaacg 480cgctcgccgc cgaagtgaac cggctcaagg gccgcgagcc
gacgcgagtg gccgagctct 540acgaggagga gctgcgggag ctgcggcgcc aggtggaggt
gctcactaac cagcgcgcgc 600gcgtcgacgt cgagcgcgac aacctgctcg acgacctgca
gcggctcaag gccaagctgc 660aggaggagat tcagttgaag gaagaagcag agaacaattt
ggctgccttc cgagcggacg 720tggatgcagc tactctagct cgcattgacc tggagcgcag
aattgaatct ctcaacgagg 780agatcgcgtt ccttaagaaa gtgcatgaag aggagatccg
tgagttgcag gctcagcttc 840aggaacagca ggtccaggtg gagatggaca tgtctaagcc
agacctcact gccgccctca 900gggatatccg ggctcagtat gagaccatcg cggctaagaa
catttctgaa gctgaggagt 960ggtacaagtc gaaggtgtca gacctgaccc aggcagccaa
caagaacaac gacgccctgc 1020gccaggccaa gcaggagatg atggaatacc gacaccagat
ccagtcctac acctgcgaga 1080ttgacgccct caagggcact aacgattccc tgatgaggca
gatgcgggaa ttggaggacc 1140gatttgccag tgaggccagt ggctaccagg acaacattgc
acgcctggag gaggaaatcc 1200ggcacctcaa ggatgagatg gcccgccatc tgcgcgagta
ccaggacctg ctcaacgtga 1260agatggccct ggatgtggag attgccacct accggaagct
gctggaggga gaggagagcc 1320ggatcaatct ccccatccag acctactctg ccctcaactt
ccgagaaacc agccctgagc 1380aaaggggttc tgaggtccat accaagaaga cggtgatgat
caagaccatc gagacacggg 1440atggggaggt cgtcagtgag gccacacagc agcagcatga
agtgctctaa agacgagaga 1500ccctctgcca ccagagaccg tcctcacccc tgtcctcact
gctccctgaa gcccagcctt 1560cttcatccca ggacaccaca cccagcctca gtcctccaaa
tcacagcctc tgacccctcc 1620tcactggcca tccctcgtgg tccccaacag cgacatagcc
catccctgcc tggtcacagg 1680catgccccgg ccacctctgc ggaccccagc tgtgagcctt
ggctgttggc agtgagtgag 1740cctggctctt gtgctggatg gagcccaggc gggagcggtg
gccctgtccc tcccacctct 1800gtgacctcag gcctacgctt tggctctgga gatagcccca
gagcagggtg ttgggatact 1860gcagggccag gactgagccc cgcagacctc cccagcccct
agcccaggag agagaaagcc 1920aggcaggtag cctgggggac tagccctgtg gagactgggg
ggcttgaaat tgtccccgtg 1980gtctcttact ttcctttccc cagcccaggg tggacttaga
aagcaggggc tacaagaggg 2040aatccccgaa ggtgctggag gtgggagcag gagattgaga
aggagagaaa gtgggtgaga 2100tgctggagaa gagaggagag gagagaggca gagagcggtc
tcaggctggt gggaggggcg 2160cccacctccc cacgccctcc ccccccctgc tgcaggggct
ctggagagaa acaataaaga 2220gattcacaca caagcc
22362566DNAHomo sapiens 2gccagaaccg gtggagcagc
gacccctgag cagtgttctc tgtgctgagc ggcgggactg 60agctgttgag ttagagccaa
catgagtgag cgacaaggtg ctggggcaac caatggaaaa 120gacaagacat ctggtgaaaa
tgatggacag aagaaagttc aagaagaatt tgacattgac 180atggatgcac cagagacaga
acgtgcagcg gtggccattc agtctcagtt cagaaaattc 240cagaagaaga aggctgggtc
tcagtcctag tgggagaacc ccctcctagt ccacctgaaa 300acaccaaatt caaccatcat
ctgtcaagaa attaaaagaa caacacccta gagagaagtc 360atccacacac aatccacaca
cgcatagcaa acctccaatg catgtacaga aacctgtgat 420atttataccc ttgtaggaag
gtatagacaa tggaattgtg agtagcttaa tctctatgtt 480tctctccatt ttcattcctc
ctgcaactat tttccttgat gttgtaataa aatgaagtta 540cgatgagtga attcaaaaaa
aaaaaa 56631433DNAHomo sapiens
3attcggggcg agggaggagg aagaagcgga ggaggcggct cccgctcgca gggccgtgca
60cctgcccgcc cgcccgctcg ctcgctcgcc cgccgcgccg cgctgccgac cgccagcatg
120ctgccgagag tgggctgccc cgcgctgccg ctgccgccgc cgccgctgct gccgctgctg
180ccgctgctgc tgctgctact gggcgcgagt ggcggcggcg gcggggcgcg cgcggaggtg
240ctgttccgct gcccgccctg cacacccgag cgcctggccg cctgcgggcc cccgccggtt
300gcgccgcccg ccgcggtggc cgcagtggcc ggaggcgccc gcatgccatg cgcggagctc
360gtccgggagc cgggctgcgg ctgctgctcg gtgtgcgccc ggctggaggg cgaggcgtgc
420ggcgtctaca ccccgcgctg cggccagggg ctgcgctgct atccccaccc gggctccgag
480ctgcccctgc aggcgctggt catgggcgag ggcacttgtg agaagcgccg ggacgccgag
540tatggcgcca gcccggagca ggttgcagac aatggcgatg accactcaga aggaggcctg
600gtggagaacc acgtggacag caccatgaac atgttgggcg ggggaggcag tgctggccgg
660aagcccctca agtcgggtat gaaggagctg gccgtgttcc gggagaaggt cactgagcag
720caccggcaga tgggcaaggg tggcaagcat caccttggcc tggaggagcc caagaagctg
780cgaccacccc ctgccaggac tccctgccaa caggaactgg accaggtcct ggagcggatc
840tccaccatgc gccttccgga tgagcggggc cctctggagc acctctactc cctgcacatc
900cccaactgtg acaagcatgg cctgtacaac ctcaaacagt gcaagatgtc tctgaacggg
960cagcgtgggg agtgctggtg tgtgaacccc aacaccggga agctgatcca gggagccccc
1020accatccggg gggaccccga gtgtcatctc ttctacaatg agcagcagga ggcttgcggg
1080gtgcacaccc agcggatgca gtagaccgca gccagccggt gcctggcgcc cctgcccccc
1140gcccctctcc aaacaccggc agaaaacgga gagtgcttgg gtggtgggtg ctggaggatt
1200ttccagttct gacacacgta tttatatttg gaaagagacc agcaccgagc tcggcacctc
1260cccggcctct ctcttcccag ctgcagatgc cacacctgct ccttcttgct ttccccgggg
1320gaggaagggg gttgtggtcg gggagctggg gtacaggttt ggggaggggg aagagaaatt
1380tttatttttg aacccctgtg tcccttttgc ataagattaa aggaaggaaa agt
143344670DNAHomo sapiens 4gcactcgctg gaaagcggct ccgagccagg ggctattgca
aagccagggt gcgctaccgg 60acggagaggg gagagccctg agcagagtga gcaacatcgc
agccaaggcg gaggccgaag 120aggggcgcca ggcaccaatc tccgcgttgc ctcagccccg
gaggcgcccc agagcgcttc 180ttgtcccagc agagccactc tgcctgcgcc tgcctctcag
tgtctccaac tttgcgctgg 240aagaaaaact tcccgcgcgc cggcagaact gcagcgcctc
cttttagtga ctccgggagc 300ttcggctgta gccggctctg cgcgcccttc caacgaataa
tagaaattgt taattttaac 360aatccagagc aggccaacga ggctttgctc tcccgacccg
aactaaaggt ccctcgctcc 420gtgcgctgct acgagcggtg tctcctgggg ctccaatgca
gcgagctgtg cccgaggggt 480tcggaaggcg caagctgggc agcgacatgg ggaacgcgga
gcgggctccg gggtctcgga 540gctttgggcc cgtacccacg ctgctgctgc tcgccgcggc
gctactggcc gtgtcggacg 600cactcgggcg cccctccgag gaggacgagg agctagtggt
gccggagctg gagcgcgccc 660cgggacacgg gaccacgcgc ctccgcctgc acgcctttga
ccagcagctg gatctggagc 720tgcggcccga cagcagcttt ttggcgcccg gcttcacgct
ccagaacgtg gggcgcaaat 780ccgggtccga gacgccgctt ccggaaaccg acctggcgca
ctgcttctac tccggcaccg 840tgaatggcga tcccagctcg gctgccgccc tcagcctctg
cgagggcgtg cgcggcgcct 900tctacctgct gggggaggcg tatttcatcc agccgctgcc
cgccgccagc gagcgcctcg 960ccaccgccgc cccaggggag aagccgccgg caccactaca
gttccacctc ctgcggcgga 1020atcggcaggg cgacgtcggc ggcacgtgcg gggtcgtgga
cgacgagccc cggccgactg 1080ggaaagcgga gaccgaagac gaggacgaag ggactgaggg
cgaggacgaa ggggctcagt 1140ggtcgccgca ggacccggca ctgcaaggcg taggacagcc
cacaggaact ggaagcataa 1200gaaagaagcg atttgtgtcc agtcaccgct atgtggaaac
catgcttgtg gcagaccagt 1260cgatggcaga attccacggc agtggtctaa agcattacct
tctcacgttg ttttcggtgg 1320cagccagatt gtacaaacac cccagcattc gtaattcagt
tagcctggtg gtggtgaaga 1380tcttggtcat ccacgatgaa cagaaggggc cggaagtgac
ctccaatgct gccctcactc 1440tgcggaactt ttgcaactgg cagaagcagc acaacccacc
cagtgaccgg gatgcagagc 1500actatgacac agcaattctt ttcaccagac aggacttgtg
tgggtcccag acatgtgata 1560ctcttgggat ggctgatgtt ggaactgtgt gtgatccgag
cagaagctgc tccgtcatag 1620aagatgatgg tttacaagct gccttcacca cagcccatga
attaggccac gtgtttaaca 1680tgccacatga tgatgcaaag cagtgtgcca gccttaatgg
tgtgaaccag gattcccaca 1740tgatggcgtc aatgctttcc aacctggacc acagccagcc
ttggtctcct tgcagtgcct 1800acatgattac atcatttctg gataatggtc atggggaatg
tttgatggac aagcctcaga 1860atcccataca gctcccaggc gatctccctg gcacctcgta
cgatgccaac cggcagtgcc 1920agtttacatt tggggaggac tccaaacact gccccgatgc
agccagcaca tgtagcacct 1980tgtggtgtac cggcacctct ggtggggtgc tggtgtgtca
aaccaaacac ttcccgtggg 2040cggatggcac cagctgtgga gaagggaaat ggtgtatcaa
cggcaagtgt gtgaacaaaa 2100ccgacagaaa gcattttgat acgccttttc atggaagctg
gggaatgtgg gggccttggg 2160gagactgttc gagaacgtgc ggtggaggag tccagtacac
gatgagggaa tgtgacaacc 2220cagtcccaaa gaatggaggg aagtactgtg aaggcaaacg
agtgcgctac agatcctgta 2280accttgagga ctgtccagac aataatggaa aaacctttag
agaggaacaa tgtgaagcac 2340acaacgagtt ttcaaaagct tcctttggga gtgggcctgc
ggtggaatgg attcccaagt 2400acgctggcgt ctcaccaaag gacaggtgca agctcatctg
ccaagccaaa ggcattggct 2460acttcttcgt tttgcagccc aaggttgtag atggtactcc
atgtagccca gattccacct 2520ctgtctgtgt gcaaggacag tgtgtaaaag ctggttgtga
tcgcatcata gactccaaaa 2580agaagtttga taaatgtggt gtttgcgggg gaaatggatc
tacttgtaaa aaaatatcag 2640gatcagttac tagtgcaaaa cctggatatc atgatatcat
cacaattcca actggagcca 2700ccaacatcga agtgaaacag cggaaccaga ggggatccag
gaacaatggc agctttcttg 2760ccatcaaagc tgctgatggc acatatattc ttaatggtga
ctacactttg tccaccttag 2820agcaagacat tatgtacaaa ggtgttgtct tgaggtacag
cggctcctct gcggcattgg 2880aaagaattcg cagctttagc cctctcaaag agcccttgac
catccaggtt cttactgtgg 2940gcaatgccct tcgacctaaa attaaataca cctacttcgt
aaagaagaag aaggaatctt 3000tcaatgctat ccccactttt tcagcatggg tcattgaaga
gtggggcgaa tgttctaagt 3060catgtgaatt gggttggcag agaagactgg tagaatgccg
agacattaat ggacagcctg 3120cttccgagtg tgcaaaggaa gtgaagccag ccagcaccag
accttgtgca gaccatccct 3180gcccccagtg gcagctgggg gagtggtcat catgttctaa
gacctgtggg aagggttaca 3240aaaaaagaag cttgaagtgt ctgtcccatg atggaggggt
gttatctcat gagagctgtg 3300atcctttaaa gaaacctaaa catttcatag acttttgcac
aatggcagaa tgcagttaag 3360tggtttaagt ggtgttagct ttgagggcaa ggcaaagtga
ggaagggctg gtgcagggaa 3420agcaagaagg ctggagggat ccagcgtatc ttgccagtaa
ccagtgaggt gtatcagtaa 3480ggtgggatta tgggggtaga tagaaaagga gttgaatcat
cagagtaaac tgccagttgc 3540aaatttgata ggatagttag tgaggattat taacctctga
gcagtgatat agcataataa 3600agccccgggc attattatta ttatttcttt tgttacatct
attacaagtt tagaaaaaac 3660aaagcaattg tcaaaaaaag ttagaactat tacaacccct
gtttcctggt acttatcaaa 3720tacttagtat catgggggtt gggaaatgaa aagtaggaga
aaagtgagat tttactaaga 3780cctgttttac tttacctcac taacaatggg gggagaaagg
agtacaaata ggatctttga 3840ccagcactgt ttatggctgc tatggtttca gagaatgttt
atacattatt tctaccgaga 3900attaaaactt cagattgttc aacatgagag aaaggctcag
caacgtgaaa taacgcaaat 3960ggcttcctct ttcctttttt ggaccatctc agtctttatt
tgtgtaattc attttgagga 4020aaaaacaact ccatgtattt attcaagtgc attaaagtct
acaatggaaa aaaagcagtg 4080aagcattaga tgctggtaaa agctagagga gacacaatga
gcttagtacc tccaacttcc 4140tttctttcct accatgtaac cctgctttgg gaatatggat
gtaaagaagt aacttgtgtc 4200tcatgaaaat cagtacaatc acacaaggag gatgaaacgc
cggaacaaaa atgaggtgtg 4260tagaacaggg tcccacaggt ttggggacat tgagatcact
tgtcttgtgg tggggaggct 4320gctgaggggt agcaggtcca tctccagcag ctggtccaac
agtcgtatcc tggtgaatgt 4380ctgttcagct cttctgtgag aatatgattt tttccatatg
tatatagtaa aatatgttac 4440tataaattac atgtacttta taagtattgg tttgggtgtt
ccttccaaga aggactatag 4500ttagtaataa atgcctataa taacatattt atttttatac
atttatttct aatgaaaaaa 4560acttttaaat tatatcgctt ttgtggaagt gcatataaaa
tagagtattt atacaatata 4620tgttactaga aataaaagaa cacttttgga aaaaaaaaaa
aaaaaaaaaa 467053360DNAHomo sapiens 5cattttactt atggaaaaca
gtgtggcata ttctgctgag cttcgccctg gaagaagcct 60cttttataca tctcttcagg
gaagagagaa gcaatgggca tgttagtata caatgatcac 120agccacgcag gcctgcaagc
tgccttttgg acaggctgtt gactgccgtt ccaattagct 180gattggagaa tgtggaatgc
agagtgataa tgctgcatat ctgctatcag gcagcagcaa 240aggtttttgt cttgggaagg
caagctttcc ctgcaatatt atctcagcag ctccctagct 300gcttaccctg aaaacgaggg
atccaaacgg agggtgttgc actctgctaa cgctggtcct 360gtgcgtggct gtggcatatg
agcggcaggt ctgaaaaagc aggtgtgtgc tgggacgggc 420actggactgg aacgcaggcg
gacgctctcg ggtttacctg cttcctgtta acagattgtg 480ggctcccagg gcatatgtct
gcacgctgag gccgaggcgg agaaggggct tcctgagcgt 540cccagtacac tgacagagac
acttggattg gacttaatct taaacctctg gagttcaaga 600ccttttaaaa agggctaaat
aaacaatctc tacatgtaaa aggccactga ctcctacttc 660ctctgtatag agcaactgtt
gaactcagct gcctgtagga aaactgaaga ctttaataac 720aaactctcca aggtgaaaat
gaacacaggg cgtgattctc agtcaccaga ctcacaaaag 780gttttagaag cgttcgacca
aacctacaag ataaaagatc accaactcag agccagataa 840cagtgaatgg aaactcagga
ggtgccgtga gtcccatgag ttactatcag aggccgtttt 900ccccctcggc atattctctc
ccagcctcac tcaactccag cattgtcatg cagcacggca 960catccctcga ttccacagac
acatatcccc agcatgcgca gtctctggat ggcaccacca 1020gcagctctat ccccctgtac
cgatcctcag aggaagagaa gagagtgaca gtcatcaaag 1080ccccgcatta cccagggatc
gggcccgtgg atgaatccgg aatccccaca gcaattagaa 1140cgacagtcga ccggcccaag
gactggtaca agacgatgtt taagcaaatt cacatggtgc 1200acaagccgga tgatgacaca
gacatgtata atactcctta tacatacaat gcaggtctgt 1260acaacccacc ctacagtgct
cagtcacacc ctgctgcaaa gacccaaacc tacagacctc 1320tttccaaaag ccactccgac
aacagcccca atgcctttaa ggatgcgtcc tccccagtgc 1380ctcccccaca tgttccacct
ccagtcccgc cgcttcgacc aagagatcgg tcttcaacag 1440aaaagcatga ctgggatcct
ccagacagaa aagtggacac aagaaaattt cggtctgagc 1500caaggagtat ttttgaatat
gaacctggca agtcatcaat tcttcagcat gaaagaccaa 1560ctgatcgcat aaatccagat
gacatagatt tagaaaatga gccctggtat aaattctttt 1620cagaactgga gtttggacgc
ccgcctccta aaaagcctct ggactatgtt caagatcatt 1680cttctggtgt tttcaatgag
gcctccttgt atcagtcctc tatagacaga agcctggaaa 1740gacccatgag ttctgcaagc
atggccagtg acttcaggaa gcggaggaag agcgagcctg 1800cagtgggtcc accacggggc
ttgggagatc aaagtgcgag caggactagc ccaggccgag 1860tggacctccc aggatcaagc
accactctta caaagtcttt cactagctct tctccttctt 1920ccccatcaag agcaaaagac
cgtgagtccc ctagaagtta ctcatccact ttgactgaca 1980tggggagaag tgcaccaagg
gaaagaagag gaactccaga aaaagagaaa ttgcctgcaa 2040aagctgttta tgattttaag
gctcagacat ctaaggagtt gtcatttaag aaaggagata 2100ctgtctacat cctcaggaaa
attgatcaaa attggtatga gggagaacac cacgggagag 2160tgggcatctt cccgatctca
tacgtagaga aactcacacc tcctgagaaa gcacagcctg 2220caagaccacc tccgccagcc
cagcccggag aaatcggaga agctatagcc aaatacaact 2280tcaacgcaga cacaaatgtg
gagctgtcac tgagaaaggg agatagagtt attcttctta 2340aaagagttga tcaaaactgg
tatgaaggta aaatcccagg aaccaacaga caaggcatct 2400tccctgtttc ctatgtggag
gtcgtcaaga agaacacaaa aggtgctgag gactaccctg 2460accctccaat accccacagc
tattctagtg ataggattca cagcttgagc tcaaataagc 2520cacagcgtcc tgtgtttact
catgaaaata ttcaaggtgg gggggaaccg tttcaggctc 2580tgtataacta tactcccagg
aatgaagatg agctggagct cagagaaagt gatgtcattg 2640atgtcatgga aaagtgtgat
gacggctggt ttgtggggac ctcaagaaga accaaattct 2700ttggtacttt ccccggaaac
tacgtcaaga ggctgtgaat tgcgctccct ccttctgtag 2760aggccgcctg ccagccatgc
acctgcgtca acgcgcctga aacaccccgc gggcctcccg 2820ttgtcatgcc ttacggtttc
caatgcgccg tcaccatctc cacctgccac caaaccacca 2880gcagagtagc cgccgctgct
gtgagcctgg ggacgacatg gcaggctggt ccccctccgt 2940gaaagtgtgg attcctactt
cctgctctaa gctttgacac gtcaaaatgt gggatcagaa 3000agaaaaaaat catgatattt
aaaaatggtc aaatatttga ggcaaaaaaa aaaaaaaaaa 3060gtgtctccag gaggctgtcc
agcctcgtgg ctccatttca acatctcccc ccaggcgatg 3120ttctccccca agacgaccag
aaaattgttt attggggaat gctgtggttt gcattttcat 3180attcttcgct tggcagtgtg
tattcttttc acaagtttgc ctagtgtctt ggtttacaca 3240atatgacaac tgtaactgta
ctttagctat tgtttgcctg cacatacatg ttgtaatatg 3300cacagtgatt acaaccttta
aagcaagagg aggcgagtta atttggatga gtgtgatttt 33606724DNAHomo sapiens
6gtagccggac cctttgcctt cgccactgct cagcgtctgc acatccctac aatggctaaa
60acagcaatgg cctacaagga aaaaatgaag gagctgtcca tgctgtcact gatctgctct
120tgcttttacc cggaacctcg caacatcaac atctatactt acgatgatat ggaagtgaag
180caaatcaaca aacgtgcctc tggccaggct tttgagctga tcttgaagcc accatctcct
240atctcagaag ccccacgaac tttagcttct ccaaagaaga aagacctgtc cctggaggag
300atccagaaga aactggaggc tgcagaggaa agaagaaagt ctcaggaggc ccaggtgctg
360aaacaattgg cagagaagag ggaacacgag cgagaagtcc ttcagaaggc tttggaggag
420aacaacaact tcagcaagat ggcggaggaa aagctgatcc tgaaaatgga acaaattaag
480gaaaaccgtg aggctaatct agctgctatt attgaacgtc tgcaggaaaa ggagaggcat
540gctgcggagg tgcgcaggaa caaggaactc caggttgaac tgtctggctg aagcaaggga
600gggtctggca cgccccacca atagtaaatc cccctgccta tattataatg gatcatgcga
660tatcaggatg gggaatgtat gacatggttt aaaaagaact cattataaaa aaaaaaaaaa
720aaaa
72472961DNAHomo sapiens 7aagagatgat ttctccatcc tgaacgtgca gcgagcttgt
caggaagatc ggaggtgcca 60agtagcagag aaagcatccc ccagctctga cagggagaca
gcacatgtct aaggcccaca 120agccttggcc ctaccggagg agaagtcaat tttcttctcg
aaaatacctg aaaaaagaaa 180tgaattcctt ccagcaacag ccaccgccat tcggcacagt
gccaccacaa atgatgtttc 240ctccaaactg gcagggggca gagaaggacg ctgctttcct
cgccaaggac ttcaactttc 300tcactttgaa caatcagcca ccaccaggaa acaggagcca
accaagggca atggggcccg 360agaacaacct gtacagccag tacgagcaga aggtgcgccc
ctgcattgac ctcatcgact 420ccctgcgggc tctgggtgtg gagcaggacc tggccctgcc
agccatcgcc gtcatcgggg 480accagagctc gggcaagagc tctgtgctgg aggcactgtc
aggagtcgcg cttcccagag 540gcagcggaat cgtaaccagg tgtccgctgg tgctgaaact
gaaaaagcag ccctgtgagg 600catgggccgg aaggatcagc taccggaaca ccgagctaga
gcttcaggac cctggccagg 660tggagaaaga gatacacaaa gcccagaacg tcatggccgg
gaatggccgg ggcatcagcc 720atgagctcat cagcctggag atcacctccc ctgaggttcc
agacctgacc atcattgacc 780ttcccggcat caccagggtg gctgtggaca accagccccg
agacatcgga ctgcagatca 840aggctctcat caagaagtac atccagaggc agcagacgat
caacttggtg gtggttccct 900gtaacgtgga cattgccacc acggaggcgc tgagcatggc
ccatgaggtg gacccggaag 960gggacaggac catcggtatc ctgaccaaac cagatctaat
ggacaggggc actgagaaaa 1020gcgtcatgaa tgtggtgcgg aacctcacgt accccctcaa
gaagggctac atgattgtga 1080agtgccgggg ccagcaggag atcacaaaca ggctgagctt
ggcagaggca accaagaaag 1140aaattacatt ctttcaaaca catccatatt tcagagttct
cctggaggag gggtcagcca 1200cggttccccg actggcagaa agacttacca ctgaactcat
catgcatatc caaaaatcgc 1260tcccgttgtt agaaggacaa ataagggaga gccaccagaa
ggcgaccgag gagctgcggc 1320gttgcggggc tgacatcccc agccaggagg ccgacaagat
gttctttcta attgagaaaa 1380tcaagatgtt taatcaggac atcgaaaagt tagtagaagg
agaagaagtt gtaagggaga 1440atgagacccg tttatacaac aaaatcagag aggattttaa
aaactgggta ggcatacttg 1500caactaatac ccaaaaagtt aaaaatatta tccacgaaga
agttgaaaaa tatgaaaagc 1560agtatcgagg caaggagctt ctgggatttg tcaactacaa
gacatttgag atcatcgtgc 1620atcagtacat ccagcagctg gtggagcccg cccttagcat
gctccagaaa gccatggaaa 1680ttatccagca agctttcatt aacgtggcca aaaaacattt
tggcgaattt ttcaacctta 1740accaaactgt tcagagcacg attgaagaca taaaagtgaa
acacacagca aaggcagaaa 1800acatgatcca acttcagttc agaatggagc agatggtttt
ttgtcaagat cagatttaca 1860gtgttgttct gaagaaagtc cgagaagaga tttttaaccc
tctggggacg ccttcacaga 1920atatgaagtt gaactctcat tttcccagta atgagtcttc
ggtttcctcc tttactgaaa 1980taggcatcca cctgaatgcc tacttcttgg aaaccagcaa
acgtctcgcc aaccagatcc 2040catttataat tcagtatttt atgctccgag agaatggtga
ctccttgcag aaagccatga 2100tgcagatact acaggaaaaa aatcgctatt cctggctgct
tcaagagcag agtgagaccg 2160ctaccaagag aagaatcctt aaggagagaa tttaccggct
cactcaggcg cgacacgcac 2220tctgtcaatt ctccagcaaa gagatccact gaagggcggc
gatgcctgtg gttgttttct 2280tgtgcgtact cattcattct aaggggagtc ggtgcaggat
gccgcttctg ctttggggcc 2340aaactcttct gtcactatca gtgtccatct ctactgtact
ccctcagcat cagagcatgc 2400atcaggggtc cacacaggct cagctctctc caccacccag
ctcttccctg accttcacga 2460agggatggct ctccagtcct tgggtcccgt agcacacagt
tacagtgtcc taagatactg 2520ctatcattct tcgctaattt gtatttgtat tcccttcccc
ctacaagatt atgagacccc 2580agagggggaa ggtctgggtc aaattcttct tttgtatgtc
cagtctcctg cacagcacct 2640gcagcattgt aactgcttaa taaatgacat ctcactgaac
gaatgagtgc tgtgtaagtg 2700atggagatac ctgaggctat tgctcaagcc caggccttgg
acatttagtg actgttagcc 2760ggtccctttc agatccagtg gccatgcccc ctgcttccca
tggttcactg tcattgtgtt 2820tcccagcctc tccactcccc cgccagaaag gagcctgagt
gattctcttt tcttcttgtt 2880tccctgatta tgatgagctt ccattgttct gttaagtctt
gaagaggaat ttaataaagc 2940aaagaaactt tttaaaaacg t
296181890DNAHomo sapiens 8ggcacccagg gtccggcctg
cgccttcccg ccaggcctgg acactggttc aacacctgtg 60acttcatgtg tgcgcgccgg
ccacacctgc agtcacacct gtagccccct ctgccaagag 120atccataccg aggcagcgtc
ggtggctaca agccctcagt ccacacctgt ggacacctgt 180gacacctggc cacacgacct
gtggccgcgg cctggcgtct gctgcgacag gagcccttac 240ctcccctgtt ataacacctg
accgccacct aactgcccct gcagaaggag caatggcctt 300ggctcctgag agggcagccc
cacgcgtgct gttcggagag tggctccttg gagagatcag 360cagcggctgc tatgaggggc
tgcagtggct ggacgaggcc cgcacctgtt tccgcgtgcc 420ctggaagcac ttcgcgcgca
aggacctgag cgaggccgac gcgcgcatct tcaaggcctg 480ggctgtggcc cgcggcaggt
ggccgcctag cagcagggga ggtggcccgc cccccgaggc 540tgagactgcg gagcgcgccg
gctggaaaac caacttccgc tgcgcactgc gcagcacgcg 600tcgcttcgtg atgctgcggg
ataactcggg ggacccggcc gacccgcaca aggtgtacgc 660gctcagccgg gagctgtgct
ggcgagaagg cccaggcacg gaccagactg aggcagaggc 720ccccgcagct gtcccaccac
cacagggtgg gcccccaggg ccattcttgg cacacacaca 780tgctggactc caagccccag
gccccctccc tgccccagct ggtgacaagg gggacctcct 840gctccaggca gtgcaacaga
gctgcctggc agaccatctg ctgacagcgt catggggggc 900agatccagtc ccaaccaagg
ctcctggaga gggacaagaa gggcttcccc tgactggggc 960ctgtgctgga ggcccagggc
tccctgctgg ggagctgtac gggtgggcag tagagacgac 1020ccccagcccc gggccccagc
ccgcggcact aacgacaggc gaggccgcgg ccccagagtc 1080cccgcaccag gcagagccgt
acctgtcacc ctccccaagc gcctgcaccg cggtgcaaga 1140gcccagccca ggggcgctgg
acgtgaccat catgtacaag ggccgcacgg tgctgcagaa 1200ggtggtggga cacccgagct
gcacgttcct atacggcccc ccagacccag ctgtccgggc 1260cacagacccc cagcaggtag
cattccccag ccctgccgag ctcccggacc agaagcagct 1320gcgctacacg gaggaactgc
tgcggcacgt ggcccctggg ttgcacctgg agcttcgggg 1380gccacagctg tgggcccggc
gcatgggcaa gtgcaaggtg tactgggagg tgggcggacc 1440cccaggctcc gccagcccct
ccaccccagc ctgcctgctg cctcggaact gtgacacccc 1500catcttcgac ttcagagtct
tcttccaaga gctggtggaa ttccgggcac ggcagcgccg 1560tggctcccca cgctatacca
tctacctggg cttcgggcag gacctgtcag ctgggaggcc 1620caaggagaag agcctggtcc
tggtgaagct ggaaccctgg ctgtgccgag tgcacctaga 1680gggcacgcag cgtgagggtg
tgtcttccct ggatagcagc agcctcagcc tctgcctgtc 1740cagcgccaac agcctctatg
acgacatcga gtgcttcctt atggagctgg agcagcccgc 1800ctagaaccca gtctaatgag
aactccagaa agctggagca gcccacctag agctggccgc 1860ggccgcccag tctaataaaa
agaactccag 189092595DNAHomo sapiens
9tcatattagt gcatttcttt gcagaggtta cctctttttc ttgtctctcg tcaggtctct
60gacattgaca gagcctggac gttggaggaa gccccaggac gttggagggg taaagtaaaa
120gtccacagtt accgtgagag aaaaaagagg gagaaagcag tgcagccaaa ctcggaagaa
180aagagaggag gaaaaggact cgactttcac attggaacaa ccttctttcc agtgctaaag
240gatctctgat ctggggaaca acaccctgga catggctcca gagatcaact tgccgggccc
300aatgagcctc attgataaca ctaaagggca gctggtggtg aatccagaag ctctgaagat
360cctatctgca attacgcagc ctgtggtggt ggtggcgatt gtgggcctct atcgcacagg
420caaatcctac ctgatgaaca agctggctgg gaagaaaaac ggcttctctc taggctccac
480agtgaagtct cacaccaagg gaatctggat gtggtgtgtg cctcatccca agaagccaga
540acacacccta gttctgctcg acactgaggg cctgggagat atagagaagg gtgacaatga
600gaatgactcc tggatctttg ccttggccat cctcctgagc agcaccttcg tgtacaatag
660catgggaacc atcaaccagc aggccatgga ccaacttcac tatgtgacag agctgacaga
720tcgaatcaag gcaaactcct cacctggtaa caattctgta gacgactcag ctgactttgt
780gagctttttt ccagcatttg tgtggactct cagagatttc accctggaac tggaagtaga
840tggagaaccc atcactgctg atgactactt ggagctttcg ctaaagctaa gaaaaggtac
900tgataagaaa agtaaaagct ttaatgatcc tcggttgtgc atccgaaagt tcttccccaa
960gaggaagtgc ttcgtcttcg attggcccgc tcctaagaag taccttgctc acctagagca
1020gctaaaggag gaagagctga accctgattt catagaacaa gttgcagaat tttgttccta
1080catcctcagc cattccaatg tcaagactct ttcaggtggc attccagtca atgggcctcg
1140tctagagagc ctggtgctga cctacgtcaa tgccatcagc agtggggatc taccctgcat
1200ggagaacgca gtcctggcct tggcccagat agagaactca gccgcagtgg aaaaggctat
1260tgcccactat gaacagcaga tgggccagaa ggtgcagctg cccacggaaa ccctccagga
1320gctgctggac ctgcacaggg acagtgagag agaggccatt gaagtcttca tgaagaactc
1380tttcaaggat gtggaccaaa tgttccagag gaaattaggg gcccagttgg aagcaaggcg
1440agatgacttt tgtaagcaga attccaaagc atcatcagat tgttgcatgg ctttacttca
1500ggatatattt ggccctttag aagaagatgt caagcaggga acattttcta aaccaggagg
1560ttaccgtctc tttactcaga agctgcagga gctgaagaat aagtactacc aggtgccaag
1620gaaggggata caggccaaag aggtgctgaa aaaatatttg gagtccaagg aggatgtggc
1680tgatgcactt ctacagactg atcagtcact ctcagaaaag gaaaaagcga ttgaagtgga
1740acgtataaag gctgaatctg cagaagctgc aaagaaaatg ttggaggaaa tacaaaagaa
1800gaatgaggag atgatggaac agaaagagaa gagttatcag gaacatgtga aacaattgac
1860tgagaagatg gagagggaca gggcccagtt aatggcagag caagagaaga ccctcgctct
1920taaacttcag gaacaggaac gccttctcaa ggagggattc gagaatgaga gcaagagact
1980tcaaaaagac atatgggata tccagatgag aagcaaatca ttggagccaa tatgtaacat
2040actctaaaag tccaaggagc aaaatttgcc tgtccagctc cctctcccca agaaacaaca
2100tgaatgagca acttcagagt gtcaaacaac tgccattaaa cttaactcaa aatcatgatg
2160catgcatttt tgttgaacca taaagtttgc aaagtaaagg ttaagtatga ggtcaatgtt
2220ttacctacag agcaattcaa ctcatgctta tttatagtac taacttttaa tatgatcttt
2280aactaaatcc tatatttgaa atcatacaca aggactcaag agagatattg tgtaactagg
2340atgcattttc caatgagata tcttgcagtt tctgttctgg gtagattttt ttctctcata
2400tgcaccaccc ttactgtata ttcagtccta tactcttatt cagggattta actatggtcg
2460tagcataggg ctgaagtgtt gtgaatatga tgaaaatgtg atgagaccaa acaaaccatg
2520gggcacagta gagcatcact cctgccaagt ggtctttgta tggcatgctg gctgcaaata
2580aaggagatct gggac
2595102459DNAHomo sapiens 10tccttcactg aggagccaga gggaatcaat tccccaagct
gggttgagga gaacaggaga 60tggagaaagc aagtggaaga cagtccattg ctctgtccac
agtggagact ggcacagtga 120acccggggct ggagctcatg gaaaaagaag tagagcctga
gggaagcaag aggactgacg 180cacaaggaca cagcctgggg gatggactgg gcccttccac
ttaccagagg aggagtcggt 240ggcctttcag caaagcaaga agtttctgca aaacacacgc
cagattgttc aagaagatcc 300tgttgggcct gttgtgtttg gcctatgctg cctatctcct
ggcagcttgc atcttgaatt 360tccagagggc actggccttg tttgtcatca cctgcttggt
gatctttgtc ctggttcact 420cgtttttgaa aaagctcctg ggcaaaaaat taacaagatg
tctgaagccc tttgaaaact 480cccgcctgag gctttggacg aaatgggtgt ttgcaggagt
ctccttggtt ggccttatac 540tgtggttggc tttagacaca gcccaaaggc cagagcagct
gatccccttt gcaggaatct 600gcatgttcat ccttatcctc tttgcctgct ccaaacacca
cagcgcagtg tcctggagga 660cagtgttttc gggcctaggt cttcaatttg tctttgggat
cttggtcatc agaactgatc 720ttggatatac tgtatttcag tggctgggag agcaggtcca
gattttcctg aactacactg 780tggccggctc cagttttgtc tttggggata cactggtcaa
ggatgtcttt gcttttcagg 840ccttaccaat catcattttc tttggatgtg tggtgtccat
tctctactac ctgggcctgg 900tgcaatgggt agttcagaag gtcgcctggt ttttacaaat
cactatgggc accactgcta 960cagagaccct ggctgtggca ggaaacatct ttgtgggtat
gacagaggca cctctgctca 1020tccgtcccta ccttggggac atgacactct ctgaaatcca
tgcggtgatg actggagggt 1080ttgccaccat ttctggcact gtgctgggag ccttcatagc
ctttggggtt gatgcatcat 1140ccctgatttc tgcctctgtg atggccgccc cttgtgctct
cgcctcatca aagctagcgt 1200atccggaagt ggaggagtcc aagttcaaga gtgaggaggg
ggtaaagctg ccccgtggga 1260aggagaggaa tgtcctggaa gctgccagca acggagccgt
agatgccata ggccttgcta 1320ctaatgtagc agccaacctg attgcctttt tggctgtgtt
ggccttcatc aatgctgccc 1380tctcctggct gggggaattg gtggacatac aggggctcac
tttccaggtc atctgctcct 1440atctcctaag gcccatggtt ttcatgatgg gtgtagagtg
gacagactgt ccaatggtgg 1500ctgagatggt gggaatcaag ttcttcataa atgagtttgt
ggcttatcag caactgtctc 1560aatacaagaa caaacgtctc tctggaatgg aggagtggat
tgagggagag aaacagtgga 1620tttctgtgag agctgaaatc attacaacat tttcactctg
tggatttgcc aatcttagtt 1680ccataggaat cacacttgga ggcttgacat caatagtacc
tcaccggaag agtgacttgt 1740ccaaggttgt ggtcagggcc ctcttcacag gggcctgtgt
atcccttatc agtgcctgta 1800tggcaggaat cctctatgtc cccaggggag ctgaagctga
ctgtgtctcc ttcccaaaca 1860caagtttcac caatagaacc tatgagacct acatgtgctg
cagagggctc tttcagagta 1920cttctctgaa tggcaccaac cctccttctt tttctggtcc
ctgggaagat aaggagttca 1980gtgctatggc ccttactaac tgctgtggat tctacaacaa
taccgtctgt gcctaaggct 2040gcttgatcta tttctataac agttttgatc ttaaaagctt
tgtgattgca aaggtgttta 2100tgtactcagg gtgcccacaa ctcactcacc aagatgttta
acagtaagta acagtaaatg 2160taaaagattc attttgggcc gggctcagtg gctcacgcct
gtaatcccag cgctttggga 2220ggccgaggcg ggcggatcgc agggtcagga gatcgagacc
atcctggcta acacggtgaa 2280accccgtctc tactaaaggt acaaaaaatt ggccgggagt
ggtgtcgggc gactgtagtc 2340ccagctactc gcgagactga ggcaggagaa tggcgtgaat
ccgggaggcg gagcttgcag 2400cgagccggga tcgcgccact gtactccagc ctgggtgaca
gagcgagact ctgtctcag 2459113972DNAHomo sapiens 11gctgccgccg ccgcgcccgg
gcgcacccgc ccgctcgctg tcccgcgcac cccgtagcgc 60ctcgggctcc cgggccggac
agaggagcca gcccggtgcg cccctccacc tcctgctcgg 120ggggctttaa tgagacaccc
accgctgctg tggggccggc ggggagcagc accgcgacgg 180ggaccggggc tgggcgctgg
agccagaatc ggaaccacga tgtgactccg ccgccgggga 240cccgtgaggt ttgtgtggac
cccgagttcc accaggtgag aagagtgatg accatccttt 300tccttactat ggttatttca
tactttggtt gcatgaaggc tgcccccatg aaagaagcaa 360acatccgagg acaaggtggc
ttggcctacc caggtgtgcg gacccatggg actctggaga 420gcgtgaatgg gcccaaggca
ggttcaagag gcttgacatc attggctgac actttcgaac 480acgtgataga agagctgttg
gatgaggacc agaaagttcg gcccaatgaa gaaaacaata 540aggacgcaga cttgtacacg
tccagggtga tgctcagtag tcaagtgcct ttggagcctc 600ctcttctctt tctgctggag
gaatacaaaa attacctaga tgctgcaaac atgtccatga 660gggtccggcg ccactctgac
cctgcccgcc gaggggagct gagcgtgtgt gacagtatta 720gtgagtgggt aacggcggca
gacaaaaaga ctgcagtgga catgtcgggc gggacggtca 780cagtccttga aaaggtccct
gtatcaaaag gccaactgaa gcaatacttc tacgagacca 840agtgcaatcc catgggttac
acaaaagaag gctgcagggg catagacaaa aggcattgga 900actcccagtg ccgaactacc
cagtcgtacg tgcgggccct taccatggat agcaaaaaga 960gaattggctg gcgattcata
aggatagaca cttcttgtgt atgtacattg accattaaaa 1020ggggaagata gtggatttat
gttgtataga ttagattata ttgagacaaa aattatctat 1080ttgtatatat acataacagg
gtaaattatt cagttaagaa aaaaataatt ttatgaactg 1140catgtataaa tgaagtttat
acagtacagt ggttctacaa tctatttatt ggacatgtcc 1200atgaccagaa gggaaacagt
catttgcgca caacttaaaa agtctgcatt acattccttg 1260ataatgttgt ggtttgttgc
cgttgccaag aactgaaaac ataaaaagtt aaaaaaaata 1320ataaattgca tgctgcttta
attgtgaatt gataataaac tgtcctcttt cagaaaacag 1380aaaaaaaaca cacacacaca
caacaaaaat ttgaaccaaa acattccgtt tacattttag 1440acagtaagta tcttcgttct
tgttagtact atatctgttt tactgctttt aacttctgat 1500agcgttggaa ttaaaacaat
gtcaaggtgc tgttgtcatt gctttactgg cttaggggat 1560gggggatggg gggtatattt
ttgtttgttt tgtgtttttt tttcgtttgt ttgttttgtt 1620ttttagttcc cacagggagt
agagatgggg aaagaattcc tacaatatat attctggctg 1680ataaaagata catttgtatg
ttgtgaagat gtttgcaata tcgatcagat gactagaaag 1740tgaataaaaa ttaaggcaac
tgaacaaaaa aatgctcaca ctccacatcc cgtgatgcac 1800ctcccaggcc ccgctcattc
tttgggcgtt ggtcagagta agctgctttt gacggaagga 1860cctatgtttg ctcagaacac
attctttccc cccctccccc tctggtctcc tctttgtttt 1920gttttaagga agaaaaatca
gttgcgcgtt ctgaaatatt ttaccactgc tgtgaacaag 1980tgaacacatt gtgtcacatc
atgacactcg tataagcatg gagaacagtg attttttttt 2040agaacagaaa acaacaaaaa
ataaccccaa aatgaagatt attttttatg aggagtgaac 2100atttgggtaa atcatggcta
agcttaaaaa aaactcatgg tgaggcttaa caatgtcttg 2160taagcaaaag gtagagccct
gtatcaaccc agaaacacct agatcagaac aggaatccac 2220attgccagtg acatgagact
gaacagccaa atggaggcta tgtggagttg gcattgcatt 2280taccggcagt gcgggaggaa
tttctgagtg gccatcccaa ggtctaggtg gaggtggggc 2340atggtatttg agacattcca
aaacgaaggc ctctgaagga cccttcagag gtggctctgg 2400aatgacatgt gtcaagctgc
ttggacctcg tgctttaagt gcctacatta tctaactgtg 2460ctcaagaggt tctcgactgg
aggaccacac tcaagccgac ttatgcccac catcccacct 2520ctggataatt ttgcataaaa
ttggattagc ctggagcagg ttgggagcca aatgtggcat 2580ttgtgatcat gagattgatg
caatgagata gaagatgttt gctacctgaa cacttattgc 2640tttgaaacta gacttgagga
aaccagggtt tatcttttga gaacttttgg taagggaaaa 2700gggaacagga aaagaaaccc
caaactcagg ccgaatgatc aaggggaccc ataggaaatc 2760ttgtccagag acaagacttc
gggaaggtgt ctggacattc agaacaccaa gacttgaagg 2820tgccttgctc aatggaagag
gccaggacag agctgacaaa attttgctcc ccagtgaagg 2880ccacagcaac cttctgccca
tcctgtctgt tcatggagag ggtccctgcc tcacctctgc 2940cattttgggt taggagaagt
caagttggga gcctgaaata gtggttcttg gaaaaatgga 3000tccccagtga aaactagagc
tctaagccca ttcagcccat ttcacacctg aaaatgttag 3060tgatcaccac ttggaccagc
atccttaagt atcagaaagc cccaagcaat tgctgcatct 3120tagtagggtg agggataagc
aaaagaggat gttcaccata acccaggaat gaagatacca 3180tcagcaaaga atttcaattt
gttcagtctt tcatttagag ctagtctttc acagtaccat 3240ctgaatacct ctttgaaaga
aggaagactt tacgtagtgt agatttgttt tgtgttgttt 3300gaaaatatta tctttgtaat
tatttttaat atgtaaggaa tgcttggaat atctgctata 3360tgtcaacttt atgcagcttc
cttttgaggg acaaatttaa aacaaacaac cccccatcac 3420aaacttaaag gattgcaagg
gccagatctg ttaagtggtt tcataggaga cacatccagc 3480aattgtgtgg tcagtggctc
ttttacccaa taagatacat cacagtcaca tgcttgatgg 3540tttatgttga cctaagattt
attttgttaa aatctctctc tgttgtgttc gttcttgttc 3600tgttttgttt tgttttttaa
agtcttgctg tggtctcttt gtggcagaag tgtttcatgc 3660atggcagcag gcctgttgct
tttttatggc gattcccatt gaaaatgtaa gtaaatgtct 3720gtggccttgt tctctctatg
gtaaagatat tattcaccat gtaaaacaaa aaacaatatt 3780tattgtattt tagtatattt
atataattat gttattgaaa aaaattggca ttaaaactta 3840accgcatcag aacctattgt
aaatacaagt tctatttaag tgtactaatt aacatataat 3900atatgtttta aatatagaat
ttttaatgtt tttaaatata ttttcaaagt acataaaaaa 3960aaaaaaaaaa aa
3972124784DNAHomo sapiens
12gaggcgacac tgagtctcca agtccggaga ggtgcccgag ggaaaagagg cggcagctaa
60actggtcctg gagagaagcc ccttccgccc ctctcctcag ccagcatgtc ccggactccg
120ccgctcctca gtccgcgcgg tggggacccc gggccgtggc ggccggcgca gccctgacgg
180gttgcgaacc agggggcgcc ccgaacgcgg gggttggggt ctgggagcgc gagcggccgc
240tacggtacga gcggggtgtg ctgagtcccg tggccacccc cggccccagc catgaggagg
300gacgagcgag acgccaaagc catgcggtcc ctgcagccgc cggatggggc cggctcgccc
360cccgagagtc tgaggaacgg ctacgtgaag agctgcgtga gccccttgcg gcaggaccct
420ccgcgcggct tcttcttcca cctctgccgc ttctgcaacg tggagctgcg gccgccgccg
480gcctctcccc agcagccgcg gcgctgctcc cccttctgcc gggcgcgcct ctcgctgggc
540gccctggctg cctttgtcct cgccctgctg ctgggcgcgg aacccgagag ctgggctgcc
600ggggccgcct ggctgcggac gctgctgagc gtgtgttcgc acagcttgag ccccctcttc
660agcatcgcct gtgccttctt cttcctcacc tgcttcctca cccggaccaa gcggggaccc
720ggcccgggcc ggagctgcgg ctcctggtgg ctgctggcgc tgcccgcctg ctgttacctg
780ggggacttct tggtgtggca gtggtggtct tggccttggg gggatggcga cgcagggtcc
840gcggccccgc acacgccccc ggaggcggca gcgggcaggt tgctgctggt gctgagctgc
900gtagggctgc tgctgacgct cgcgcacccg ctgcggctcc ggcactgcgt tctggtgctg
960ctcctggcca gcttcgtctg gtgggtctcc ttcaccagcc tcgggtcgct gccctccgcc
1020ctcaggccgc tgctctccgg cctggtgggg ggcgctggct gcctgctggc cctggggttg
1080gatcacttct ttcaaatcag ggaagcgcct cttcatcctc gactgtccag tgccgccgaa
1140gaaaaagtgc ctgtgatccg accccggagg aggtccagct gcgtgtcgtt aggagaaact
1200gcagccagtt actatggcag ttgcaaaata ttcaggagac cgtcgttgcc ttgtatttcc
1260agagaacaga tgattctttg ggattgggac ttaaaacaat ggtataagcc tcattatcaa
1320aattctggag gtggaaatgg agttgatctt tcagtgctaa atgaggctcg caatatggtg
1380tcagatcttc tgactgatcc aagccttcca ccacaagtca tttcctctct acggagtatt
1440agtagcttaa tgggtgcttt ctcaggttcc tgtaggccaa agattaatcc tctcacacca
1500tttcctggat tttacccctg ttctgaaata gaggacccag ctgagaaagg ggatagaaaa
1560cttaacaagg gactaaatag gaatagtttg ccaactccac agctgaggag aagctcagga
1620acttcaggat tgctacctgt tgaacagtct tcaaggtggg atcgtaataa tggcaaaagg
1680cctcaccaag aatttggcat ttcaagtcaa ggatgctatc taaatgggcc ttttaattca
1740aatctactga ctatcccgaa gcaaaggtca tcttctgtat cactgactca ccatgtaggt
1800ctcagaagag ctggtgtttt gtccagtctg agtcctgtga attcttccaa ccatggacca
1860gtgtctactg gctctctaac taatcgatca cccatagaat ttcctgatac tgctgatttt
1920cttaataagc caagcgttat cttgcagaga tctctgggca atgcacctaa tactccagat
1980ttttatcagc aacttagaaa ttctgatagc aatctgtgta acagctgtgg acatcaaatg
2040ctgaaatatg tttcaacatc tgaatcagat ggtacagatt gctgcagtgg aaaatcaggt
2100gaagaagaaa acattttctc gaaagaatca ttcaaactta tggaaactca acaagaagag
2160gaaacagaga agaaagacag cagaaaatta tttcaggaag gtgataagtg gctaacagaa
2220gaggcacaga gtgaacagca aacaaatatt gaacaggaag tatcactgga cctgatttta
2280gtagaagagt atgactcatt aatagaaaag atgagcaact ggaattttcc aatttttgaa
2340cttgtagaaa agatgggaga gaaatcagga aggattctca gtcaggttat gtatacctta
2400tttcaagaca ctggtttatt ggaaatattt aaaattccca ctcaacaatt tatgaactat
2460tttcgtgcat tagaaaatgg ctatcgagac attccttatc acaatcgtat acatgccaca
2520gatgtgctac atgcagtttg gtatctgaca acacggccag ttcctggctt acagcagatc
2580cacaatggtt gtggaacagg aaatgaaaca gattctgatg gtagaattaa ccatgggcga
2640attgcttata tttcttcgaa gagctgctct aatcctgatg agagttatgg ctgcctgtct
2700tcaaacattc ctgcattaga attgatggct ctatacgtgg cagctgccat gcatgattat
2760gatcacccag ggaggacaaa tgcatttcta gtggctacaa atgcccctca ggcagtttta
2820tacaatgaca gatctgttct ggaaaatcat catgctgcgt cagcttggaa tctatatctt
2880tctcgcccag aatacaactt ccttcttcat cttgatcatg tggaattcaa gcgctttcgt
2940tttttagtca ttgaagcaat ccttgctacg gatcttaaaa agcattttga ttttctcgca
3000gaattcaatg ccaaggcaaa tgatgtaaat agtaatggca tagaatggag taatgaaaat
3060gatcgcctct tggtatgcca ggtgtgcatc aaactggcag atataaatgg cccagcaaaa
3120gttcgagact tgcatttgaa atggacagaa ggcattgtca atgaatttta tgagcaggga
3180gatgaagaag caaatcttgg tctgcccatc agtccattca tggatcgttc ttctcctcaa
3240ctagcaaaac tccaagaatc ttttatcacc cacatagtgg gtcccctgtg taactcctat
3300gatgctgctg gtttgctacc aggtcagtgg ttagaagcag aagaggataa tgatactgaa
3360agtggtgatg atgaagacgg tgaagaatta gatacagaag atgaagaaat ggaaaacaat
3420ctaaatccaa aaccaccaag aaggaaaagc agacggcgaa tattttgtca gctaatgcac
3480cacctcactg aaaaccacaa gatatggaag gaaatcgtag aggaagaaga aaaatgtaaa
3540gctgatggga ataaactgca ggtggagaat tcctccttac ctcaagcaga tgagattcag
3600gtaattgaag aggcagatga agaggaatag cgacagtttg agtaaaagaa aagtcatatt
3660gaagaagccc agagggttgt gcccaggggc agaaatcatt gcctagtgtt caccggctga
3720ctctcaactg accattccca tgtggacagg ccttaatact gtgagaggat ccttgctctg
3780ctggcagttt cccactccta tgcactttca caggaactag aaaactattc ttaaaccaaa
3840aataccatcc gtgttgaccc atgttgcaga gcccttactt aaatccttca ctggtgtatg
3900aatactttgt cataatgctg ctttgctggg tagtgagctc ttatttttca ctgggggtca
3960gctataacta aaaactcaag tgacatattt cagttaccaa agtggccagg aactttttgc
4020ttttatgaaa atagattcat attgtatttc ccagtgtgtc ttttatgtct ttgaatgttt
4080tggagaaaag tctatgcctg tctaaaaatg aatccagtgt tgcctttctg agggatttct
4140gctcaatgca atacactgtt cagtgctatt ctcccagcta ggtttatcca tgaaggactg
4200agtgaccttt gttgtattta acaaaatcca ggtgcatcaa tttctgatgc tttttactat
4260tgtgtattat ctactatgtg tgttttattt ctgctgagag tattcaggtt tgccatggac
4320atcagaagtt tgaattccag tcttatctta tgttccatgg ctgaatttta aagctgttta
4380ggtttaacaa tgaagggatt tattctttag tcaaaattgt tgtttttact ctagctcagg
4440attcgtattt ttaaagattt agttaatata aacacagcac agatttgtca gaagaaaaaa
4500aatttgctgt aataccaaaa ctaacctcat caaagataca gaaaaaaaga aatatagtga
4560gccctaaagg acacatacat tgaataaata attggaacat gtggttatct ttagatccac
4620atcttagctg tcatttgttc actctaaaac tgatgttcat ctttctgtta atttccctct
4680gcctaaagag tacatgacag aaatgaccta tcactactta ttatttctga agcctaactg
4740caagagtgat ttcttgagaa caagtaaaga actggctcgt gccg
4784132710DNAHomo sapiens 13gcaggcatcg tctcctccgg tctcacagct accccgggct
ggagcgtggg gagggggtaa 60gggagccagc caactcgggg aaacttcgtt gccagagatg
actggggttt tcgggctttc 120ccagccttcc catgccgggc cacacggcat tcattaaagt
ggtggggcaa ctattggtgc 180gctccagccg ggcaggtttt gagctgcaga gacccgtgcc
cggtagcagc gctttgccac 240cgagtgcacc caggacgcag tggccgccag cccgggtctc
ctgctggggg ccggcgagtg 300cagggagact ttgcgccacg atgttttgtt ttcctgccgc
aggaacgcta ggcagtctct 360ttccaccaga cttaacccgc ctgaagcaat agccatggaa
gttgcggcgg cacagaggga 420gacagaaccg cttggagcgg catctttctt ggatgttttt
tctctacaca gacttctttc 480tttccttggt ggccgttccc gcagcagcac cggtgtgcca
gcccaagagc gccactaacg 540ggcaaccccc ggctccggct ccgactccaa ctccgcgcct
gtccatttcc tcccgagcca 600cagtggtagc caggatggaa ggcacctccc aagggggctt
gcagaccgtc atgaagtgga 660agacggtggt tgccatcttt gtggttgtgg tggtctacct
tgtcactggc ggtcttgtct 720tccgggcatt ggagcagccc tttgagagca gccagaagaa
taccatcgcc ttggagaagg 780cggaattcct gcgggatcat gtctgtgtga gcccccagga
gctggagacg ttgatccagc 840atgctcttga tgctgacaat gcgggagtca gtccaatagg
aaactcttcc aacaacagca 900gccactggga cctcggcagt gcctttttct ttgctggaac
tgtcattacg accatagggt 960atgggaatat tgctccgagc actgaaggag gcaaaatctt
ttgtatttta tatgccatct 1020ttggaattcc actctttggt ttcttattgg ctggaattgg
agaccaactt ggaaccatct 1080ttgggaaaag cattgcaaga gtggagaagg tctttcgaaa
aaagcaagtg agtcagacca 1140agatccgggt catctcaacc atcctgttca tcttggccgg
ctgcattgtg tttgtgacga 1200tccctgctgt catctttaag tacatcgagg gctggacggc
cttggagtcc atttactttg 1260tggtggtcac tctgaccacg gtgggctttg gtgattttgt
ggcaggggga aacgctggca 1320tcaattatcg ggagtggtat aagcccctag tgtggttttg
gatccttgtt ggccttgcct 1380actttgcagc tgtcctcagt atgatcggag attggctacg
ggttctgtcc aaaaagacaa 1440aagaagaggt gggtgaaatc aaggcccatg cggcagagtg
gaaggccaat gtcacggctg 1500agttccggga gacacggcga aggctcagcg tggagatcca
cgataagctg cagcgggcgg 1560ccaccatccg cagcatggag cgccggcggc tgggcctgga
ccagcgggcc cactcactgg 1620acatgctgtc ccccgagaag cgctctgtct ttgctgccct
ggacaccggc cgcttcaagg 1680cctcatccca ggagagcatc aacaaccggc ccaacaacct
gcgcctgaag gggccggagc 1740agctgaacaa gcatgggcag ggtgcgtccg aggacaacat
catcaacaag ttcgggtcca 1800cctccagact caccaagagg aaaaacaagg acctcaaaaa
gaccttgccc gaggacgttc 1860agaaaatcta caagaccttc cggaattact ccctggacga
ggagaagaaa gaggaggaga 1920cggaaaagat gtgtaactca gacaactcca gcacagccat
gctgacggac tgtatccagc 1980agcacgctga gttggagaac ggaatgatac ccacggacac
caaagaccgg gagccggaga 2040acaactcatt acttgaagac agaaactaaa tgtgaaggac
attggtcttg gactgagcgt 2100tgtgtgtgtg tgtgtgtgtg tgtttttaat attcacactg
agacatgtgc cttaaacaga 2160ctttttagtc caaaattaca tagcattgaa gaatatattt
cactgtgcca taaacaactg 2220aaagcttgct ctgccaaaag gaatcagaga acaagaactt
catttcagat agcaaacgca 2280ggacacacca agagtgtccg tgcacgtagc cggttctggc
cgtacatgtt aagggcattt 2340cagtggcagt gctgtacccc tgggcagtgc tacctgggca
cacacgtaga caagggcagc 2400tattccttag accagcctcc tgaaagaaac aggtgtgtct
ttttagtgga gtcgtagtaa 2460tatgtgcaca cacagaaggg gacctgattg ggtgggagct
ggttatgtgt aactagcgtt 2520ggagttgaca ttttggcatg tgctctgagc ttgaattttg
ataccaacca ttcagtgcat 2580catacctagt ctttctatgc tccaaatgaa tgtctgtggg
gacctgagag cacctggaat 2640ttgttggaag cagatcagag cacacgtacg aaaaggtgca
attgcttttc tcatgacaaa 2700agaaaaaaaa
2710142391DNAHomo sapiens 14atgctgcgag gcggacggcg
cgggcagctt ggctggcaca gctgggctgc ggggccgggc 60agcctgctgg cttggctgat
actggcatct gcgggcgccg caccctgccc cgatgcctgc 120tgcccccacg gctcctcggg
actgcgatgc acccgggatg gggccctgga tagcctccac 180cacctgcccg gcgcagagaa
cctgactgag ctctacatcg agaaccagca gcatctgcag 240catctggagc tccgtgatct
gaggggcctg ggggagctga gaaacctcac catcgtgaag 300agtggtctcc gtttcgtggc
gccagatgcc ttccatttca ctcctcggct cagtcgcctg 360aatctctcct tcaacgctct
ggagtctctc tcctggaaaa ctgtgcaggg cctctcctta 420caggaactgg tcctgtcggg
gaaccctctg cactgttctt gtgccctgcg ctggctacag 480cgctgggagg aggagggact
gggcggagtg cctgaacaga agctgcagtg tcatgggcaa 540gggcccctgg cccacatgcc
caatgccagc tgtggtgtgc ccacgctgaa ggtccaggtg 600cccaatgcct cggtggatgt
gggggacgac gtgctgctgc ggtgccaggt ggaggggcgg 660ggcctggagc aggccggctg
gatcctcaca gagctggagc agtcagccac ggtgatgaaa 720tctgggggtc tgccatccct
ggggctgacc ctggccaatg tcaccagtga cctcaacagg 780aagaacgtga cgtgctgggc
agagaacgat gtgggccggg cagaggtctc tgttcaggtc 840aacgtctcct tcccggccag
tgtgcagctg cacacggcgg tggagatgca ccactggtgc 900atccccttct ctgtggatgg
gcagccggca ccgtctctgc gctggctctt caatggctcc 960gtgctcaatg agaccagctt
catcttcact gagttcctgg agccggcagc caatgagacc 1020gtgcggcacg ggtgtctgcg
cctcaaccag cccacccacg tcaacaacgg caactacacg 1080ctgctggctg ccaacccctt
cggccaggcc tccgcctcca tcatggctgc cttcatggac 1140aaccctttcg agttcaaccc
cgaggacccc atccctgtct ccttctcgcc ggtggacact 1200aacagcacat ctggagaccc
ggtggagaag aaggacgaaa caccttttgg ggtctcggtg 1260gctgtgggcc tggccgtctt
tgcctgcctc ttcctttcta cgctgctcct tgtgctcaac 1320aaatgtggac ggagaaacaa
gtttgggatc aaccgcccgg ctgtgctggc tccagaggat 1380gggctggcca tgtccctgca
tttcatgaca ttgggtggca gctccctgtc ccccaccgag 1440ggcaaaggct ctgggctcca
aggccacatc atcgagaacc cacaatactt cagtgatgcc 1500tgtgttcacc acatcaagcg
ccgggacatc gtgctcaagt gggagctggg ggagggcgcc 1560tttgggaagg tcttccttgc
tgagtgccac aacctcctgc ctgagcagga caagatgctg 1620gtggctgtca aggcactgaa
ggaggcgtcc gagagtgctc ggcaggactt ccaacgtgag 1680gctgagctgc tcaccatgct
gcagcaccag cacatcgtgc gcttcttcgg cgtctgcacc 1740gagggccgcc ccctgctcat
ggtcttcgag tatatgcggc acggggacct caaccgcttc 1800ctccgatccc atggacccga
tgccaagctg ctggctggtg gggaggatgt ggctccaggc 1860cccctgggtc tggggcagct
gctggccgtg gctagccagg tcgctgcggg gatggtgtac 1920ctggcgggtc tgcattttgt
gcaccgggac ctggccacac gcaactgtct agtgggccag 1980ggactggtgg tcaagattgg
tgattttggc atgagcaggg atatctacag caccgactat 2040taccgtgtgg gaggccgcac
catgctgccc attcgctgga tgccgcccga gagcatcctg 2100taccgtaagt tcaccaccga
gagcgacgtg tggagcttcg gcgtggtgct ctgggagatc 2160ttcacctacg gcaagcagcc
ctggtaccag ctctccaaca cggaggcaat cgactgcatc 2220acgcagggac gtgagttgga
gcggccacgt gcctgcccac cagaggtcta cgccatcatg 2280cggggctgct ggcagcggga
gccccagcaa cgccacagca tcaaggatgt gcacgcccgg 2340ctgcaagccc tggcccaggc
acctcctgtc tacctggatg tcctgggcta g 2391154909DNAHomo sapiens
15tagacgcacc ctctgaagat ggtgactccc tcctgagaag ctggacccct tggtaaaaga
60caaggccttc tccaagaaga atatgaaagt gttactcaga cttatttgtt tcatagctct
120actgatttct tctctggagg ctgataaatg caaggaacgt gaagaaaaaa taattttagt
180gtcatctgca aatgaaattg atgttcgtcc ctgtcctctt aacccaaatg aacacaaagg
240cactataact tggtataaag atgacagcaa gacacctgta tctacagaac aagcctccag
300gattcatcaa cacaaagaga aactttggtt tgttcctgct aaggtggagg attcaggaca
360ttactattgc gtggtaagaa attcatctta ctgcctcaga attaaaataa gtgcaaaatt
420tgtggagaat gagcctaact tatgttataa tgcacaagcc atatttaagc agaaactacc
480cgttgcagga gacggaggac ttgtgtgccc ttatatggag ttttttaaaa atgaaaataa
540tgagttacct aaattacagt ggtataagga ttgcaaacct ctacttcttg acaatataca
600ctttagtgga gtcaaagata ggctcatcgt gatgaatgtg gctgaaaagc atagagggaa
660ctatacttgt catgcatcct acacatactt gggcaagcaa tatcctatta cccgggtaat
720agaatttatt actctagagg aaaacaaacc cacaaggcct gtgattgtga gcccagctaa
780tgagacaatg gaagtagact tgggatccca gatacaattg atctgtaatg tcaccggcca
840gttgagtgac attgcttact ggaagtggaa tgggtcagta attgatgaag atgacccagt
900gctaggggaa gactattaca gtgtggaaaa tcctgcaaac aaaagaagga gtaccctcat
960cacagtgctt aatatatcgg aaattgaaag tagattttat aaacatccat ttacctgttt
1020tgccaagaat acacatggta tagatgcagc atatatccag ttaatatatc cagtcactaa
1080tttccagaag cacatgattg gtatatgtgt cacgttgaca gtcataattg tgtgttctgt
1140tttcatctat aaaatcttca agattgacat tgtgctttgg tacagggatt cctgctatga
1200ttttctccca ataaaagctt cagatggaaa gacctatgac gcatatatac tgtatccaaa
1260gactgttggg gaagggtcta cctctgactg tgatattttt gtgtttaaag tcttgcctga
1320ggtcttggaa aaacagtgtg gatataagct gttcatttat ggaagggatg actacgttgg
1380ggaagacatt gttgaggtca ttaatgaaaa cgtaaagaaa agcagaagac tgattatcat
1440tttagtcaga gaaacatcag gcttcagctg gctgggtggt tcatctgaag agcaaatagc
1500catgtataat gctcttgttc aggatggaat taaagttgtc ctgcttgagc tggagaaaat
1560ccaagactat gagaaaatgc cagaatcgat taaattcatt aagcagaaac atggggctat
1620ccgctggtca ggggacttta cacagggacc acagtctgca aagacaaggt tctggaagaa
1680tgtcaggtac cacatgccag tccagcgacg gtcaccttca tctaaacacc agttactgtc
1740accagccact aaggagaaac tgcaaagaga ggctcacgtg cctctcgggt agcatggaga
1800agttgccaag agttctttag gtgcctcctg tcttatggcg ttgcaggcca ggttatgcct
1860catgctgact tgcagagttc atggaatgta actatatcat cctttatccc tgaggtcacc
1920tggaatcaga ttattaaggg aataagccat gacgtcaata gcagcccagg gcacttcaga
1980gtagagggct tgggaagatc ttttaaaaag gcagtaggcc cggtgtggtg gctcacgcct
2040ataatcccag cactttggga ggctgaagtg ggtggatcac cagaggtcag gagttcgaga
2100ccagcccagc caacatggca aaaccccatc tctactaaaa atacaaaaat gagctaggca
2160tggtggcaca cgcctgtaat cccagctaca cctgaggctg aggcaggaga attgcttgaa
2220ccggggagac ggaggttgca gtgagccgag tttgggccac tgcactctag cctggcaaca
2280gagcaagact ccgtctcaaa aaaagggcaa taaatgccct ctctgaatgt ttgaactgcc
2340aagaaaaggc atggagacag cgaactagaa gaaagggcaa gaaggaaata gccaccgtct
2400acagatggct tagttaagtc atccacagcc caagggcggg gctatgcctt gtctggggac
2460cctgtagagt cactgaccct ggagcggctc tcctgagagg tgctgcaggc aaagtgagac
2520tgacacctca ctgaggaagg gagacatatt cttggagaac tttccatctg cttgtatttt
2580ccatacacat ccccagccag aagttagtgt ccgaagaccg aattttattt tacagagctt
2640gaaaactcac ttcaatgaac aaagggattc tccaggattc caaagttttg aagtcatctt
2700agctttccac aggagggaga gaacttaaaa aagcaacagt agcagggaat tgatccactt
2760cttaatgctt tcctccctgg catgaccatc ctgtcctttg ttattatcct gcattttacg
2820tctttggagg aacagctccc tagtggcttc ctccgtctgc aatgtccctt gcacagccca
2880cacatgaacc atccttccca tgatgccgct cttctgtcat cccgctcctg ctgaaacacc
2940tcccaggggc tccacctgtt caggagctga agcccatgct ttcccaccag catgtcactc
3000ccagaccacc tccctgccct gtcctccagc ttcccctcgc tgtcctgctg tgtgaattcc
3060caggttggcc tggtggccat gtcgcctgcc cccagcactc ctctgtctct gctcttgcct
3120cgacccttcc tcctcctttg cctaggaggc cttctcgcat tttctctagc tgatcagaat
3180tttaccaaaa ttcagaacat cctccaattc cacagtctct gggagacttt ccctaagagg
3240cgacttcctc tccagccttc tctctctggt caggcccact gcagagatgg tggtgagcac
3300atctgggagg ctggtctccc tccagctgga attgctgctc tctgagggag aggctgtggt
3360ggctgtctct gtccctcact gccttccagg agcaatttgc acatgtaaca tagatttatg
3420taatgcttta tgtttaaaaa cattccccaa ttatcttatt taatttttgc aattattcta
3480attttatata tagagaaagt gacctatttt ttaaaaaaat cacactctaa gttctattga
3540acctaggact tgagcctcca tttctggctt ctagtctggt gttctgagta cttgatttca
3600ggtcaataac ggtcccccct cactccacac tggcacgttt gtgagaagaa atgacatttt
3660gctaggaagt gaccgagtct aggaatgctt ttattcaaga caccaaattc caaacttcta
3720aatgttggaa ttttcaaaaa ttgtgtttag attttatgaa aaactcttct actttcatct
3780attctttccc tagaggcaaa catttcttaa aatgtttcat tttcattaaa aatgaaagcc
3840aaatttatat gccaccgatt gcaggacaca agcacagttt taagagttgt atgaacatgg
3900agaggacttt tggtttttat atttctcgta tttaatatgg gtgaacacca acttttattt
3960ggaataataa ttttcctcct aaacaaaaac acattgagtt taagtctctg actcttgcct
4020ttccacctgc tttctcctgg gcccgctttg cctgcttgaa ggaacagtgc tgttctggag
4080ctgctgttcc aacagacagg gcctagcttt catttgacac acagactaca gccagaagcc
4140catggagcag ggatgtcacg tcttgaaaag cctattagat gttttacaaa tttaattttg
4200cagattattt tagtctgtca tccagaaaat gtgtcagcat gcatagtgct aagaaagcaa
4260gccaatttgg aaacttaggt tagtgacaaa attggccaga gagtgggggt gatgatgacc
4320aagaattaca agtagaatgg cagctggaat ttaaggaggg acaagaatca atggataagc
4380gtgggtggag gaagatccaa acagaaaagt gcaaagttat tccccatctt ccaagggttg
4440aattctggag gaagaagaca cattcctagt tccccgtgaa cttcctttga cttattgtcc
4500ccactaaaac aaaacaaaaa acttttaatg ccttccacat taattagatt ttcttgcagt
4560ttttttatgg cattttttta aagatgccct aagtgttgaa gaagagtttg caaatgcaac
4620aaaatattta attaccggtt gttaaaactg gtttagcaca atttatattt tccctctctt
4680gcctttctta tttgcaataa aaggtattga gccatttttt aaatgacatt tttgataaat
4740tatgtttgta ctagttgatg aaggagtttt ttttaacctg tttatataat tttgcagcag
4800aagccaaatt ttttgtatat taaagcacca aattcatgta cagcatgcat cacggatcaa
4860tagactgtac ttattttcca ataaaatttt caaactttgt actgttaaa
4909163599DNAHomo sapiens 16ggcccctgcg cccttcctgg gatcactccg actgccccgc
gcctctgcca actcctctgg 60accctcgcgg ccgtgggcag cggctgccgc gcctgtctgc
ccgagggagg accttcgcct 120ctgcatttgt ccagtaactc tggctgtgcc ggatactgct
tgggtaaaac gggcacccca 180ggaacatggc agacgaagat ctcatcttcc gcctggaagg
cgttgatggc ggccagtccc 240cccgagctgg ccatgatggt gattctgatg gggacagcga
cgatgaggaa ggttacttca 300tctgccccat cacggatgac ccaagctcga accagaatgt
caattccaag gttaataagt 360actacagcaa cctaacaaaa agtgagcggt atagctccag
cgggtccccg gcaaactcct 420tccacttcaa ggaagcctgg aagcacgcaa tccagaaggc
caagcacatg cccgacccct 480gggctgagtt ccacctggaa gatattgcca ccgaacgtgc
tactcgacac aggtacaacg 540ccgtcaccgg ggaatggctg gatgatgaag ttctgatcaa
gatggcatct cagcccttcg 600gccgaggagc aatgagggag tgcttccgga cgaagaagct
ctccaacttc ttgcatgccc 660agcagtggaa gggcgcctcc aactacgtgg cgaagcgcta
catcgagccc gtagaccggg 720atgtgtactt tgaggacgtg cgtctacaga tggaggccaa
gctctggggg gaggagtata 780atcggcacaa gccccccaag caggtggaca tcatgcagat
gtgcatcatc gagctgaagg 840acagaccggg caagcccctc ttccacctgg agcactacat
cgagggcaag tacatcaagt 900acaactccaa ctctggcttt gtccgcgatg acaacatccg
cctgacgccg caggccttca 960gccacttcac ttttgagcgt tccggccatc agctgatagt
ggtggacatc cagggagttg 1020gggatctcta cactgaccca cagatccaca cggagacggg
cactgacttt ggagacggca 1080acctaggtgt ccgcgggatg gcgctcttct tctactctca
tgcctgcaac cggatttgcg 1140agagcatggg ccttgctccc tttgacctct cgccccggga
gagggatgca gtgaatcaga 1200acaccaagct gctgcaatca gccaagacca tcttgagagg
aacagaggaa aaatgtggga 1260gcccccgagt aaggaccctc tctgggagcc ggccacccct
gctccgtccc ctttcagaga 1320actctggaga cgagaacatg agcgacgtga ccttcgactc
tctcccttct tccccatctt 1380cggccacacc acacagccag aagctagacc acctccattg
gccagtgttc agtgacctcg 1440ataacatggc atccagagac catgatcatc tagacaacca
ccgggagtct gagaatagtg 1500gggacagcgg ataccccagt gagaagcggg gtgagctgga
tgaccctgag ccccgagaac 1560atggccactc atacagtaat cggaagtacg agtctgacga
agacagcctg ggcagctctg 1620gacgggtatg tgtagagaag tggaatctcc tcaactcctc
ccgcctccac ctgccgaggg 1680cttcggccgt ggccctggaa gtgcaaaggc ttaatgctct
ggacctcgaa aagaaaatcg 1740ggaagtccat tttggggaag gtccatctgg ccatggtgcg
ctaccacgag ggtgggcgct 1800tctgcgagaa gggcgaggag tgggaccagg agtcggctgt
cttccacctg gagcacgcag 1860ccaacctggg cgagctggag gccatcgtgg gcctgggact
catgtactcg cagttgcctc 1920atcacatcct agccgatgtc tctctgaagg agacagaaga
gaacaaaacc aaaggatttg 1980attacttact aaaggccgct gaagctggcg acaggcagtc
catgatccta gtggcgcgag 2040cttttgactc tggccagaac ctcagcccgg acaggtgcca
agactggcta gaggccctgc 2100actggtacaa cactgccctg gagatgacgg actgtgatga
gggcggtgag tacgacggaa 2160tgcaggacga gccccggtac atgatgctgg ccagggaggc
cgagatgctg ttcacaggag 2220gctacgggct ggagaaggac ccgcagagat caggggactt
gtatacccag gcagcagagg 2280cagcgatgga agccatgaag ggccgactgg ccaaccagta
ctaccaaaag gctgaagagg 2340cctgggccca gatggaggag taaccaggaa aatcactgcc
ggctagtccc aagcaaacgg 2400gctaggagga aagattaaaa aaacaacaac aacaacttat
ttagtttggg gaggggaagc 2460atttttaagt gtgttgtaaa atcaaatttt atatttcatt
ttttgactct tgaaaaatgt 2520ctttgctcct tggcagctac cagcagagac tctatagctg
tctcttaggg cagtattttg 2580gggaagtggg gcttgaagaa gcagcctaat gaaccaacat
accgttttgt gtgtggtttt 2640ttttgtttgt ttgtttgttt gttttgagac agagtcttgc
tctgtcaccc aggctggagt 2700gcagtgacat gatcttagct cactgcaacc tccgcctcct
gggttcaagt gattctcctg 2760cctcagcctc ccaagtagct gggattactg gtgcacacca
ccacactcag ctaatttttg 2820catttttagt agagatgggg tttcaccatg ttggccaggc
tggtctcgaa ctcctggact 2880taagtgagcc tcccgcctca gtctcccaaa gcgctgggat
tacaggcagg agccactgag 2940cccagccaag acttcagtgt tgactgcttt ggaggcacaa
acccatgcaa gcgttagttc 3000caaagttcag tgtgtaccct taaatgaaca atgaagcagg
taaaattacc cttgaaaaaa 3060atcccttgga ccacccataa atgacagtga ctttttcaat
atggactcat catagccagt 3120tttccttttg aagttggaac tgatcaccct tttgtcatct
gtaccagatc agtagttggc 3180ttgtgttaca ttttgtgtgt gtgtgtgcgt gttttaaacc
agtgcatata aattgtatgt 3240taaatgtaag taactttaag ttgacttatc tcttcacagt
aatcaagcct cacgtaattc 3300atgcttttta aattcagcca gccccccctc tctgaaattt
tattatgtaa ataatttgtg 3360ttccctgatc actcgtttaa gttcttagtt gtatgtcatc
tcttctctag caggaattgg 3420caaacttttt tgtaaagggg tagaaagtga agattttagg
ctttgcaggc catatagcct 3480ctgctgcaaa tgctcagccc tgctgttgta atgtaaaagc
tgccacagac actacatgaa 3540cacgaatgag tgtggctggt gttccaataa aactttattt
acacaaaaaa aaaaaaaaa 3599171288DNAHomo sapiens 17acaccagccc tcagtcactg
ggagaagaac ctctcatacc ctcggtgctc cagtccccag 60ctcactcagc cacacacacc
atgtgtgaag aggagaccac cgcgctcgtg tgtgacaatg 120gctctggcct gtgcaaggca
ggcttcgcag gagatgatgc cccccgggct gtcttcccct 180ccattgtggg ccgccctcgc
caccagggtg tgatggtggg aatgggccag aaagacagct 240atgtggggga tgaggctcag
agcaagcgag ggatcctaac tctcaaatac cccattgaac 300acggcatcat caccaactgg
gatgacatgg agaagatctg gcaccactcc ttctacaatg 360agctgcgtgt agcacctgaa
gagcacccca ccctgctcac agaggctccc ctaaatccca 420aggccaacag ggaaaagatg
acccagatca tgtttgaaac cttcaatgtc cctgccatgt 480acgtcgccat tcaagctgtg
ctctccctct atgcctctgg ccgcacgaca ggcatcgtcc 540tggattcagg tgatggcgtc
acccacaatg tccccatcta tgaaggctat gccctgcccc 600atgccatcat gcgcctggac
ttggctggcc gtgacctcac ggactacctc atgaagatcc 660tcacagagag aggctattcc
tttgtgacca cagctgagag agaaattgtg cgagacatca 720aggagaagct gtgctatgtg
gccctggatt ttgagaatga gatggccaca gcagcttcct 780cttcctccct ggagaagagc
tatgagctgc cagatgggca ggttatcacc attggcaatg 840agcgcttccg ctgccctgag
accctcttcc agccttcctt tattggcatg gagtccgctg 900gaattcatga gacaacctac
aattccatca tgaagtgtga cattgacatc cgtaaggact 960tatatgccaa caatgtcctc
tctgggggca ccaccatgta ccctggcatt gctgacagga 1020tgcagaagga gatcacagcc
ctggccccca gcaccatgaa gatcaagatt attgctcccc 1080cagagcggaa gtactcagtc
tggatcgggg gctctatcct ggcctctctc tccaccttcc 1140agcagatgtg gatcagcaag
cctgagtatg atgaggcagg gccctccatt gtccacagga 1200agtgcttcta aagtcagaac
aggttctcca aggatcccct cgagactact ctgttaccag 1260tcatgaaaca ttaaaaccta
caagcctt 1288186861DNAHomo sapiens
18gcctgggagg tgcgtcagat ccgagctcgc catccagttt cctctccact agtcccccca
60gttggagatc tgggaccaac aaggcaccat ggcgcagaag ggccaactca gtgacgatga
120gaagttcctc tttgtggaca aaaacttcat caacagccca gtggcccagg ctgactgggc
180cgccaagaga ctcgtctggg tcccctcgga gaagcagggc ttcgaggcag ccagcattaa
240ggaggagaag ggggatgagg tggttgtgga gctggtggag aatggcaaga aggtcacggt
300tgggaaagat gacatccaga agatgaaccc acccaagttc tccaaggtgg aggacatggc
360ggagctgacg tgcctcaacg aagcctccgt gctacacaac ctgagggagc ggtacttctc
420agggctaata tatacgtact ctggcctctt ctgcgtggtg gtcaacccct ataaacacct
480gcccatctac tcggagaaga tcgtcgacat gtacaagggc aagaagaggc acgagatgcc
540gcctcacatc tacgccatcg cagacacggc ctaccggagc atgcttcaag atcgggagga
600ccagtccatt ctatgcacag gcgagtctgg agccgggaaa accgaaaaca ccaagaaggt
660cattcagtac ctggccgtgg tggcctcctc ccacaagggc aagaaagaca caagtatcac
720gggagagctg gaaaagcagc ttctacaagc aaacccgatt ctggaggctt tcggcaacgc
780caaaacagtg aagaacgaca actcctcacg attcggcaaa ttcatccgca tcaacttcga
840cgtcacgggt tacatcgtgg gagccaacat tgagacctat ctgctagaaa aatcacgggc
900aattcgccaa gccagagacg agaggacatt ccacatcttt tactacatga ttgctggagc
960caaggagaag atgagaagtg acttgctttt ggagggcttc aacaactaca ccttcctctc
1020caatggcttt gtgcccatcc cagcagccca ggatgatgag atgttccagg aaaccgtgga
1080ggccatggca atcatgggtt tcagcgagga ggagcagcta tccatattga aggtggtatc
1140atcggtcctg cagcttggaa atatcgtctt caagaaggaa agaaacacag accaggcgtc
1200catgccagat aacacagctg ctcagaaagt ttgccacctc atgggaatta atgtgacaga
1260tttcaccaga tccatcctca ctcctcgtat caaggttggg cgagatgtgg tacagaaagc
1320tcagacaaaa gaacaggctg actttgctgt agaggctttg gccaaggcaa catatgagcg
1380ccttttccgc tggatactca cccgcgtgaa caaagccctg gacaagaccc atcggcaagg
1440ggcttccttc ctggggatcc tggatatagc tggatttgag atctttgagg tgaactcctt
1500cgagcagctg tgcatcaact acaccaacga gaagctgcag cagctcttca accacaccat
1560gttcatcctg gagcaggagg agtaccagcg cgagggcatc gagtggaact tcatcgactt
1620tgggctggac ctacagccct gcatcgagct catcgagcga ccgaacaacc ctccaggtgt
1680gctggccctg ctggacgagg aatgctggtt ccccaaagcc acggacaagt ctttcgtgga
1740gaagctgtgc acggagcagg gcagccaccc caagttccag aagcccaagc agctcaagga
1800caagactgag ttctccatca tccattatgc tgggaaggtg gactataatg cgagtgcctg
1860gctgaccaag aatatggacc cgctgaatga caacgtgact tccctgctca atgcctcctc
1920cgacaagttt gtggccgacc tgtggaagga cgtggaccgc atcgtgggcc tggaccagat
1980ggccaagatg acggagagct cgctgcccag cgcctccaag accaagaagg gcatgttccg
2040cacagtgggg cagctgtaca aggagcagct gggcaagctg atgaccacgc tacgcaacac
2100cacgcccaac ttcgtgcgct gcatcatccc caaccacgag aagaggtccg gcaagctgga
2160tgcgttcctg gtgctggagc agctgcggtg caatggggtg ctggaaggca ttcgcatctg
2220ccggcagggc ttccccaacc ggatcgtctt ccaggagttc cgccaacgct acgagatcct
2280ggcggcgaat gccatcccca aaggcttcat ggacgggaag caggcctgca ttctcatgat
2340caaagccctg gaacttgacc ccaacttata caggataggg cagagcaaaa tcttcttccg
2400aactggcgtc ctggcccacc tagaggagga gcgagatttg aagatcaccg atgtcatcat
2460ggccttccag gcgatgtgtc gtggctactt ggccagaaag gcttttgcca agaggcagca
2520gcagctgacc gccatgaagg tgattcagag gaactgcgcc gcctacctca agctgcggaa
2580ctggcagtgg tggaggcttt tcaccaaagt gaagccactg ctgcaggtga cacggcagga
2640ggaggagatg caggccaagg aggatgaact gcagaagacc aaggagcggc agcagaaggc
2700agagaatgag cttaaggagc tggaacagaa gcactcgcag ctgaccgagg agaagaacct
2760gctacaggaa cagctgcagg cagagacaga gctgtatgca gaggctgagg agatgcgggt
2820gcggctggcg gccaagaagc aggagctgga ggagatactg catgagatgg aggcccgcct
2880ggaggaggag gaagacaggg gccagcagct acaggctgaa aggaagaaga tggcccagca
2940gatgctggac cttgaagaac agctggagga ggaggaagct gccaggcaga agctgcaact
3000tgagaaggtc acggctgagg ccaagatcaa gaaactggag gatgagatcc tggtcatgga
3060tgatcagaac aataaactat caaaagaacg aaaactcctt gaggagagga ttagtgactt
3120aacgacaaat cttgcagaag aggaagaaaa ggccaagaat cttaccaagc tgaaaaacaa
3180gcatgaatct atgatttcag aactggaagt gcggctaaag aaggaagaga agagccgaca
3240ggagctggag aagctgaaac ggaagctgga gggtgatgcc agcgacttcc acgagcagat
3300cgctgacctc caggcgcaga tcgcagagct caagatgcag ctggccaaga aggaggagga
3360gctgcaggcg gccctggcca ggcttgacga tgaaatcgct cagaagaaca atgccctgaa
3420gaagatccgg gagctggagg gccacatctc agacctccag gaggacctgg actcagagcg
3480ggccgccagg aacaaggctg aaaagcagaa gcgagacctc ggcgaggagc tggaggccct
3540aaagacagag ctggaagaca cactggacag cacagccact cagcaggagc tcagggccaa
3600gagggagcag gaggtgacgg tgctgaagaa ggccctggat gaagagacgc ggtcccatga
3660ggctcaggtc caggagatga ggcagaaaca cgcacaggcg gtggaggagc tcacagagca
3720gcttgagcag ttcaagaggg ccaaggcgaa cctagacaag aataagcaga cgctggagaa
3780agagaacgca gacctggccg gggagctgcg ggtcctgggc caggccaagc aggaggtgga
3840acataagaag aagaagctgg aggcgcaggt gcaggagctg cagtccaagt gcagcgatgg
3900ggagcgggcc cgggcggagc tcaatgacaa agtccacaag ctgcagaatg aagttgagag
3960cgtcacaggg atgcttaacg aggccgaggg gaaggccatt aagctggcca aggacgtggc
4020gtccctcagt tcccagctcc aggacaccca ggagctgctt caagaagaaa cccggcagaa
4080gctcaacgtg tctacgaagc tgcgccagct ggaggaggag cggaacagcc tgcaagacca
4140gctggacgag gagatggagg ccaagcagaa cctggagcgc cacatctcca ctctcaacat
4200ccagctctcc gactcgaaga agaagctgca ggactttgcc agcaccgtgg aagctctgga
4260agaggggaag aagaggttcc agaaggagat cgagaacctc acccagcagt acgaggagaa
4320ggcggccgct tatgataaac tggaaaagac caagaacagg cttcagcagg agctggacga
4380cctggttgtt gatttggaca accagcggca actcgtgtcc aacctggaaa agaagcagag
4440gaaatttgat cagttgttag ccgaggagaa aaacatctct tccaaatacg cggatgagag
4500ggacagagct gaggcagaag ccagggagaa ggaaaccaag gccctgtccc tggctcgggc
4560ccttgaagag gccttggaag ccaaagagga actcgagcgg accaacaaaa tgctcaaagc
4620cgaaatggaa gacctggtca gctccaagga tgacgtgggc aagaacgtcc atgagctgga
4680gaagtccaag cgggccctgg agacccagat ggaggagatg aagacgcagc tggaagagct
4740ggaggacgag ctgcaagcca cggaggacgc caaactgcgg ctggaagtca acatgcaggc
4800gctcaagggc cagttcgaaa gggatctcca agcccgggac gagcagaatg aggagaagag
4860gaggcaactg cagagacagc ttcacgagta tgagacggaa ctggaagacg agcgaaagca
4920acgtgccctg gcagctgcag caaagaagaa gctggaaggg gacctgaaag acctggagct
4980tcaggccgac tctgccatca aggggaggga ggaagccatc aagcagctac gcaaactgca
5040ggctcagatg aaggactttc aaagagagct ggaagatgcc cgtgcctcca gagatgagat
5100ctttgccaca gccaaagaga atgagaagaa agccaagagc ttggaagcag acctcatgca
5160gctacaagag gacctcgccg ccgctgagag ggctcgcaaa caagcggacc tcgagaagga
5220ggaactggca gaggagctgg ccagtagcct gtcgggaagg aacgcactcc aggacgagaa
5280gcgccgcctg gaggcccgga tcgcccagct ggaggaggag ctggaggagg agcagggcaa
5340catggaggcc atgagcgacc gggtccgcaa agccacacag caggccgagc agctcagcaa
5400cgagctggcc acagagcgca gcacggccca gaagaatgag agtgcccggc agcagctcga
5460gcggcagaac aaggagctcc ggagcaagct ccacgagatg gagggggccg tcaagtccaa
5520gttcaagtcc accatcgcgg cgctggaggc caagattgca cagctggagg agcaggtcga
5580gcaggaggcc agagagaaac aggcggccac caagtcgctg aagcagaaag acaagaagct
5640gaaggaaatc ttgctgcagg tggaggacga gcgcaagatg gccgagcagt acaaggagca
5700ggcagagaaa ggcaatgcca gggtcaagca gctcaagagg cagctggagg aggcagagga
5760ggagtcccag cgcatcaacg ccaaccgcag gaagctgcag cgggagctgg atgaggccac
5820ggagagcaac gaggccatgg gccgcgaggt gaacgcactc aagagcaagc tcaggcgagg
5880aaacgagacc tctttcgttc cttctagaag gtctggagga cgtagagtta ttgaaaatgc
5940agatggttct gaggaggaaa cggacactcg agacgcagac ttcaatggaa ccaaggccag
6000tgaataagca actttctaca gttttgcacc acggcaagaa aaccaaaaac caaaacaaac
6060aaacaaaaaa aacccaacaa caacccagaa caaagcaaaa cccagcagac tgtacttagc
6120attgtctaaa tccattctca aattccaaat atcacagaca cccctcacac aaggaatata
6180aaaaccacca ccctccagcc tgggcaacgt agtaaaacct catctataca agaatttaaa
6240aataagctgg gcgtggtggt acacacctgt ggtcccagct actagggagg ctgagccagg
6300aagaacgctc cagcccagga cttcgaggct gcaatgagct ataattgcat cattgcactc
6360cagcctgggc aacagagacc ctgtctcaac caccaccacc accaccaccc ctactacccc
6420tgtattcaag gtaaaaattg aagtttgtat gatgtaagag atgagaaaaa cccaacagga
6480aacacagaca catcctccag ttctatcaat ggattgtgca gacactgagt ttttagaaaa
6540acatatccac ggtaaccggt ccctggcaat tctgtttaca tgaaatgggg agaaagtcac
6600cgaaatgggt gccgccggcc cccactccca attcattccc taacctgcaa acctttccaa
6660cttctcacgt caggcctttg agaattcttt ccccctctcc tggtttccac acctcagaca
6720cgcacagttc accaagtgcc ttctgtagtc acatgaattg aaaaggagac gctgctccca
6780cggaggggag caggaatgct gcactgttta caccctgact gtgcttaaaa acactttcac
6840taataaatgg ttataaatca c
6861191243DNAHomo sapiens 19ggacgcaacc atctgcggaa aggatagggg ggaggaagac
ttaacactgc aacaacaggt 60acaaaaaaga aggaaactca gtaacaggac ccagaggacc
attatatatt gctctatatt 120ctaggtcgcc cactttacac ttccttctca tgcacttggt
caataccacg cccgctgacc 180acactggcga cttccctctc tgtcgcccct ccgtgaagtc
agacccactc tgcgggccaa 240gaaaggtgac cgggcttcct tccggcttgc taagcagagg
ccggaagcgg tggtttttag 300cggctctctg ggtagcaggg tggtgtgata gcggcagcga
ggggctcgga gaggtgctcg 360gattctcgta gctgtgccgg gacttaacca ccaccatgtc
gagcaaaaga acaaagacca 420agaccaagaa gcgccctcag cgtgcaacat ccaatgtgtt
tgctatgttt gaccagtcgc 480agattcagga gttcaaagag gccttcaaca tgattgatca
gaacagagat ggtttcatcg 540acaaggaaga tttgcatgat atgcttgctt cattggggaa
gaatccaact gatgagtatc 600tagatgccat gatgaatgag gctccaggcc ccatcaattt
caccatgttc ctcaccatgt 660ttggtgagaa gttaaatggc acagatcctg aagatgtcat
cagaaatgcc tttgcttgct 720ttgatgaaga agcaactggc accatacagg aagattactt
gagagagctg ctgacaacca 780tgggggatcg gtttacagat gaggaagtgg atgagctgta
cagagaagca cctattgata 840aaaaggggaa tttcaattac atcgagttca cacgcatcct
gaaacatgga gccaaagaca 900aagatgactg aaataacttc aaattccagc caaacgttcc
ttgttgccac tttgggtatt 960ctgagatttt ctcttgcatg cccttagctt tacagctttt
gcatttcctg ttgtatttat 1020tctcagccat tttgggcata tgtatcttta taatcagact
ggaaacggga ctttctatta 1080atatcatttt cagaataaaa aataggataa tttaacctac
cagcccttct cccccaataa 1140ctgtggtcta tacagagtca atatattttt tcggagaaag
ttattcgctc gattttttct 1200gaatcataat taaactttat gataaaataa aaaaaaaaaa
aaa 124320878DNAHomo sapiens 20aggaagtgtc ggcgccgcca
ctgtccggcc acagcctaac gctcttcgct gtcgtttgcg 60gtctcggcag ggcgcccccg
ttctggtgtt tggcgtcgga attaaacaac caccatgtcg 120agcaaaaagg caaagaccaa
gaccaccaag aagcgccctc agcgtgcaac atccaatgtg 180tttgccatgt ttgaccagtc
acagattcag gagttcaaag aggccttcaa catgattgat 240cagaacagag atggcttcat
cgacaaggaa gatttgcatg atatgcttgc ttctctaggg 300aagaatccca ctgatgcata
ccttgatgcc atgatgaatg aggccccagg gccaatcaat 360ttcaccatgt tcctgaccat
gtttggtgag aagttaaatg gcacagatcc tgaagatgtc 420atcagaaacg cctttgcttg
ctttgatgaa gaagcaacag gtaccattca ggaagattac 480ctaagagagc tgctgacaac
catgggggat cggtttacag atgaggaagt ggatgagctg 540tacagagaag cacctattga
caaaaagggg aatttcaatt acatcgagtt cacacgcatc 600ctgaaacatg gagccaaaga
caaagatgac tgaaagaact ttagctaaaa tcttccagtt 660acattgtctt actctctttt
acttctcaga cacttccccc accctcatag aacctgttgc 720atgcaactta gtttcacagc
tttgcctctt ctttttgatg tatttattcc agacctttct 780gccacttagc acttgtataa
tcagactgga aatggggatg agggtgtaaa ttgtattgaa 840aaagatcgcg aataaaaatc
aacaaagtgt gaaagccc 878211681DNAHomo sapiens
21agttggaggg aggcagggaa tctggcttga ttggcgtgct gagacgcacc tggcgcaacc
60ctcccttctg aatcgaagtt caagtcccgc ggacactgca accatgaagg agagacgggc
120cccccagcca gtcgtggcca gatgtaagct cgttctggtc ggggacgtgc agtgtgggaa
180gaccgcgatg ttgcaagtgt tagcgaagga ttgctatcca gagacctatg tgcccaccgt
240gttcgaaaat tacacagcct gtttggagac agaggaacag agggtggagc ttagtctctg
300ggatacctca ggatctccct actacgataa tgtccgtcca ctctgctaca gcgactcgga
360tgcagtatta ctatgttttg acatcagccg tccagagaca gtggacagcg cactcaagaa
420gtggaggaca gaaatcctag attattgtcc cagcacccgc gttttgctca ttggctgcaa
480gacagacctg cgaacagacc tgagtactct gatggagctg tcccaccaga agcaggcgcc
540catctcctat gagcagggtt gtgcaatagc aaagcagctg ggtgcagaaa tctacctgga
600aggctcagct ttcacctcag aaaagagcat ccacagcatc tttcggacgg catccatgct
660gtgtctgaac aagcctagcc cactgcccca gaagagccct gtccgaagcc tctccaaacg
720actgctccac ctccccagtc gctctgaact catctcttct accttcaaga aggaaaaggc
780caaaagctgt tccattatgt gaagtggaaa ttggaggggg gagacaaccc cctacttcct
840cccttggggt gcagaggcac ggggagaggg aggatgagac aatttaggac actggacatg
900agtttttcag atggccacgg tgagggcttg gaaggagaca ggaatggggc gaggaaggag
960ccaggcccgg catgaggacc tgacgctgag agagaaccat cataccccaa gccaggcact
1020agattttgga gggggcgact accccagtgc cccccccgct ccagaggaag gaaagctgtg
1080ggggacgggg ggcatgctgg cctcatgggc ttgggggcct acagcagcct caccttcagc
1140ttcatgcctc ttccacacag cgtttccatg caggtcaggg gatgggaggg gtccctgagc
1200ccttcccttc ccctctaagg aggcagcaac ggagagtggg gaagtggagc ggcagctccc
1260ttgggggctt agcccaggtg cttcgtaact gcaatcggaa gtgcaggggc tggtcagagc
1320caatgagaag gaaacctcat ctttgcatag cccatgcctc atggagaggt gacatcatac
1380attcacatgc ttctcaccta agtccccagg gtccaaggga gaagccccag acccccttct
1440cttgcagagt gtgggggtgg tggtgctgca ggggcagggc tgggtggggg tcaccagact
1500ttttctgccc ttagggtagt acagctggca tttgttttat agactcttgt ctttggaatt
1560ggggggaggg ggggagtgtt tcaatctgtt atatgttctg tgtttaatga agaaaaccta
1620tttattaatg aaaaatataa tacatataaa gaaaaaaaaa aaaaagaaaa aaaaaaaaaa
1680a
168122865DNAHomo sapiens 22ctcaaacacc gcctgctaaa aatacccgac tggaggagca
taaaagcgca gccgagccca 60gcgccccgca cttttctgag cagacgtcca gagcagagtc
agccagcatg accgagcgcc 120gcgtcccctt ctcgctcctg cggggcccca gctgggaccc
cttccgcgac tggtacccgc 180atagccgcct cttcgaccag gccttcgggc tgccccggct
gccggaggag tggtcgcagt 240ggttaggcgg cagcagctgg ccaggctacg tgcgccccct
gccccccgcc gccatcgaga 300gccccgcagt ggccgcgccc gcctacagcc gcgcgctcag
ccggcaactc agcagcgggg 360tctcggagat ccggcacact gcggaccgct ggcgcgtgtc
cctggatgtc aaccacttcg 420ccccggacga gctgacggtc aagaccaagg atggcgtggt
ggagatcacc ggcaagcacg 480aggagcggca ggacgagcat ggctacatct cccggtgctt
cacgcggaaa tacacgctgc 540cccccggtgt ggaccccacc caagtttcct cctccctgtc
ccctgagggc acactgaccg 600tggaggcccc catgcccaag ctagccacgc agtccaacga
gatcaccatc ccagtcacct 660tcgagtcgcg ggcccagctt gggggcccag aagctgcaaa
atccgatgag actgccgcca 720agtaaagcct tagcctggat gcccacccct gctgccgcca
ctggctgtgc ctcccccgcc 780acctgtgtgt tcttttgata catttatctt ctgtttttct
caaataaagt tcaaagcaac 840cacctgtaaa aaaaaaaaaa aaaaa
865233890DNAHomo sapiens 23gcacctgggc acctgggcag
ccgccgcggc gctggctaga cgtgcgcgat ggagggcgac 60ggcgggaccc catgggccct
ggcgctgctg cgcaccttcg acgcgggcga gttcacgggc 120tgggagaagg tgggctcggg
cggcttcggg caggtgtaca aggtgcgcca tgtccactgg 180aagacctggc tggccatcaa
gtgctcgccc agcctgcacg tcgacgacag ggagcgcatg 240gagcttttgg aagaagccaa
gaagatggag atggccaagt ttcgctacat cctgcctgtg 300tatggcatct gccgcgaacc
tgtcggcctg gtcatggagt acatggagac gggctccctg 360gaaaagctgc tggcttcgga
gccattgcca tgggatctcc ggttccgaat catccacgag 420acggcggtgg gcatgaactt
cctgcactgc atggccccgc cactcctgca cctggacctc 480aagcccgcga acatcctgct
ggatgcccac taccacgtca agatttctga ttttggtctg 540gccaagtgca acgggctgtc
ccactcgcat gacctcagca tggatggcct gtttggcaca 600atcgcctacc tccctccaga
gcgcatcagg gagaagagcc ggctcttcga caccaagcac 660gatgtataca gctttgcgat
cgtcatctgg ggcgtgctca cacagaagaa gccgtttgca 720gatgagaaga acatcctgca
catcatggtg aaggtggtga agggccaccg ccccgagctg 780ccgcccgtgt gcagagcccg
gccgcgcgcc tgcagccacc tgatacgcct catgcagcgg 840tgctggcagg gggatccgcg
agttaggccc accttccaag aaattacttc tgaaaccgag 900gacctgtgtg aaaagcctga
tgacgaagtg aaagaaactg ctcatgatct ggacgtgaaa 960agccccccgg agcccaggag
cgaggtggtg cctgcgaggc tcaagcgggc ctctgccccc 1020accttcgata acgactacag
cctctccgag ctgctctcac agctggactc tggagtttcc 1080caggctgtcg agggccccga
ggagctcagc cgcagctcct ctgagtccaa gctgccatcg 1140tccggcagtg ggaagaggct
ctcgggggtg tcctcggtgg actccgcctt ctcttccaga 1200ggatcactgt cgctgtcctt
tgagcgggaa ccttcaacca gcgatctggg caccacagac 1260gtccagaaga agaagcttgt
ggatgccatc gtgtccgggg acaccagcaa actgatgaag 1320atcctgcagc cgcaggacgt
ggacctggca ctggacagcg gtgccagcct gctgcacctg 1380gcggtggagg ccgggcaaga
ggagtgcgcc aagtggctgc tgctcaacaa tgccaacccc 1440aacctgagca accgtagggg
ctccaccccg ttgcacatgg ccgtggagag gagggtgcgg 1500ggtgtcgtgg agctcctgct
ggcgcggaag atcagtgtca acgccaagga tgaggaccag 1560tggacagccc tccactttgc
agcccagaac ggggacgagt ctagcacacg gctgctgttg 1620gagaagaacg cctcggtcaa
cgaggtggac tttgagggcc ggacgcccat gcacgtggcc 1680tgccagcacg ggcaggagaa
tatcgtgcgc atcctgctgc gccgaggcgt ggacgtgagc 1740ctgcagggca aggatgcctg
gctgccactg cactacgctg cctggcaggg ccacctgccc 1800atcgtcaagc tgctggccaa
gcagccgggg gtgagtgtga acgcccagac gctggatggg 1860aggacgccat tgcacctggc
cgcacagcgc gggcactacc gcgtggcccg catcctcatc 1920gacctgtgct ccgacgtcaa
cgtctgcagc ctgctggcac agacacccct gcacgtggcc 1980gcggagacgg ggcacacgag
cactgccagg ctgctcctgc atcggggcgc tggcaaggag 2040gccatgacct cagacggcta
caccgctctg cacctggctg cccgcaacgg acacctggcc 2100actgtcaagc tgcttgtcga
ggagaaggcc gatgtgctgg cccggggacc cctgaaccag 2160acggcgctgc acctggctgc
cgcccacggg cactcggagg tggtggagga gttggtcagc 2220gccgatgtca ttgacctgtt
cgacgagcag gggctcagcg cgctgcacct ggccgcccag 2280ggccggcacg cacagacggt
ggagactctg ctcaggcatg gggcccacat caacctgcag 2340agcctcaagt tccagggcgg
ccatggcccc gccgccacgc tcctgcggcg aagcaagacc 2400tagctggctg cctgcggaga
ccgggggtcc acgtggggct cttgtcctgt cctgtgttcc 2460tcgtggggat ggaacgatcc
tgcgtggggc cctgttgtgg cttacctaaa tgttaaccaa 2520gcagaggtga catggtgcca
tcaggaggcg gctgctgctg accggagtgt cccctccagg 2580tgaagctggc tcaggtgcac
atgcccgctc catcatcgat ctaggcacct gctgtctgaa 2640gggaccgtgg gtcagaatca
tttcgttgtg ctcctaatgg gtcgctgagg ctggtctctc 2700agtgatgaag ccccaggcgt
ggaagcatcc actctctcct gaggcgagcc accttgggtt 2760gctggagctc accagtcttg
agggaggtgc aggggaaact gtgtttttta tcttcataca 2820tgacggtggg cagagaggcc
tgtcttaaag tttccatgga attgttttat aaaatatctt 2880aagagatgaa taccttatca
gctgttgctt gaaacctgtt aaaaatgttc ataacattgg 2940atagtctagt ctctaaatga
tgactaagta gtggggttgg ctttgaaaac aatgttttat 3000gcaacaagga acgaatggta
gcagccagct ttgcggggcg tatgtgtggc cagctcttaa 3060ccattccagt ctattacttg
ggtgagtcct tgtggacaac cacacacacg tgcccacatg 3120gtactagctg ccgttcgttt
ctcgttgcct aagatgtttt ggcaactcta gagccacagg 3180cctaagtcat taaaaaattc
tccctttgta acctcagtgc tggggactga ggcgagcccc 3240ctcaggtcgc tggagtgcac
cagtcttggg gaagaggtgc aggagaagct gtgtttttta 3300tctccacacg cagtatgaag
ataaaattac atagtattac ctagacatag acagtattac 3360ctaggtagat gcactgctca
cctgcgccct tcccagctct catttttgtt aggtgatttg 3420ggatagggat agtgttttgg
ggtatggggg gagtgtttct gacctgcttt gcagacgtgc 3480ctccgcacct cagcagtttg
gggtgtggcc ccagggcggt tcttggatgt aaaagatgtg 3540gccatctagc ctcgtaactt
cactgtcacc tgtgtcccat agggtgcctt ctgaatactg 3600ttattagaat aagtttgttg
cagaacgtga ccctgcgtgc aaacatgtac cgtggcctgg 3660tatatgatag agattgatat
taatgtacca tgtatgttaa tgtgaatctg tgggcaggat 3720acttttccat ggcaggaaat
atccaagctg ttgaaactgg ctatgtttta atatgcctca 3780ttgtgccttt actgttgtgt
ggactgcgtg agggacaaga agttccattt gatgtcaata 3840aagcaaagta cttgcctact
tttttgaagc tgaaaaaaaa aaaaaaaaaa 389024894DNAHomo sapiens
24cgtcatacgc agcgcccttt gtgacaccag ggccctggtg ctttaactag ggcgttggga
60cctgttgccc acacagaccg ccctgcagtt tcagactgga gggcggtgga cggctactca
120gcggcccaac tctctcgcag cccttctctc cgcaaaatgt cagcctccac ctcctcgcac
180cggcccatca aggggatcct gaaaaacaaa agctcgtcgg gttcctcggt ggcgacttcc
240ggtcagcagt ctggagggac tattcaagat gtgaagagaa agaaatccca aaagtgggac
300gaatcaagca tccttgcggc acaccgcgca acgtacagag attacgattt aatgaaggca
360aatgagcccg gcacttccta catgagtgtg caagataatg gggaagattc agtgcgcgat
420gtcgaaggag aagattcagt gcgtggtgtc gaaggaaagg aagccaccga tgcttccgac
480cacagctgtg aggtggacga gcaagagagc agtgaggcct acatgagaaa aatcctcctc
540cacaaacagg agaaaaagcg gcagttcgaa atgagaagaa ggcttcacta caacgaagaa
600ttgaacatca aattagctag acaattaatg tggaaagagc tacaaagtga agataatgaa
660aacgaagaaa cgccacaagg cacgaacgaa gagaagactg ctgcggaaga atcagaggaa
720gctcctctga ccggtggact gcaaacccag tcatgcgacc cttagaagat gcctgcttca
780cccttgcaat tgtttgtgaa tatgtgacgc ttagaagata tctgcttcac ccttgcaatt
840gtttgtgaaa tacaaacctt gttactgtaa aaaaaaaaaa aaaaaaaaaa aaaa
894251574DNAHomo sapiens 25tcaccacggc ggcagccctt taaacccctc acccagccag
cgccccatcc tgtctgtccg 60aacccagaca caagtcttca ctccttcctg cgagccctga
ggaagccttg tgagtgcatt 120ggctggggct tggagggaag ttgggctgga gctggacagg
agcagtgggt gcatttcagg 180caggctctcc tgaggtccca ggcgccagct ccagctccct
ggctagggaa acccaccctc 240tcagtcagca tgggggccca agctccaggc agggtgggct
ggatcactag cgtcctggat 300ctctctcaga ctgggcagcc ccgggctcat tgaaatgccc
cggatgactt ggctagtgca 360gaggaattga tggaaaccac cggggtgaga gggaggctcc
ccatctcagc cagccacatc 420cacaaggtgt gtgtaagggt gcaggcgccg gccggttagg
ccaaggctct actgtctgtt 480gcccctccag gagaacttcc aaggagcttt ccccagacat
ggccaacaag ggtccttcct 540atggcatgag ccgcgaagtg cagtccaaaa tcgagaagaa
gtatgacgag gagctggagg 600agcggctggt ggagtggatc atagtgcagt gtggccctga
tgtgggccgc ccagaccgtg 660ggcgcttggg cttccaggtc tggctgaaga atggcgtgat
tctgagcaag ctggtgaaca 720gcctgtaccc tgatggctcc aagccggtga aggtgcccga
gaacccaccc tccatggtct 780tcaagcagat ggagcaggtg gctcagttcc tgaaggcggc
tgaggactat ggggtcatca 840agactgacat gttccagact gttgacctct ttgaaggcaa
agacatggca gcagtgcaga 900ggaccctgat ggctttgggc agcttggcag tgaccaagaa
tgatgggcac taccgtggag 960atcccaactg gtttatgaag aaagcgcagg agcataagag
ggaattcaca gagagccagc 1020tgcaggaggg aaagcatgtc attggccttc agatgggcag
caacagaggg gcctcccagg 1080ccggcatgac aggctacgga cgacctcggc agatcatcag
ttagagcgga gagggctagc 1140cctgagcccg gccctccccc agctccttgg ctgcagccat
cccgcttagc ctgcctcacc 1200cacacccgtg tggtaccttc agccctggcc aagctttgag
gctctgtcac tgagcaatgg 1260taactgcacc tgggcagctc ctccctgtgc ccccagcctc
agcccaactt cttacccgaa 1320agcatcactg ccttggcccc tccctcccgg ctgcccccat
cacctctact gtctcctccc 1380tgggctaagc aggggagaag cgggctgggg gtagcctgga
tgtgggccaa gtccactgtc 1440ctccttggcg gcaaaagccc attgaagaag aaccagccca
gcctgccccc tatcttgtcc 1500tggaatattt ttggggttgg aactcaaaaa aaaaaaaaaa
aaatcaatct tttctcaaaa 1560aaaaaaaaaa aaaa
1574263700DNAHomo sapiens 26agccgccgcc acccgccgcg
cccgacaccc gggaggcccc gccagcccgc gggagaggcc 60cagcgggagt cgcggaacag
caggcccgag cccaccgcgc cgggccccgg acgccgcgcg 120gaaaagatga atttacaacc
aattttctgg attggactga tcagttcagt ttgctgtgtg 180tttgctcaaa cagatgaaaa
tagatgttta aaagcaaatg ccaaatcatg tggagaatgt 240atacaagcag ggccaaattg
tgggtggtgc acaaattcaa catttttaca ggaaggaatg 300cctacttctg cacgatgtga
tgatttagaa gccttaaaaa agaagggttg ccctccagat 360gacatagaaa atcccagagg
ctccaaagat ataaagaaaa ataaaaatgt aaccaaccgt 420agcaaaggaa cagcagagaa
gctcaagcca gaggatatta ctcagatcca accacagcag 480ttggttttgc gattaagatc
aggggagcca cagacattta cattaaaatt caagagagct 540gaagactatc ccattgacct
ctactacctt atggacctgt cttactcaat gaaagacgat 600ttggagaatg taaaaagtct
tggaacagat ctgatgaatg aaatgaggag gattacttcg 660gacttcagaa ttggatttgg
ctcatttgtg gaaaagactg tgatgcctta cattagcaca 720acaccagcta agctcaggaa
cccttgcaca agtgaacaga actgcaccag cccatttagc 780tacaaaaatg tgctcagtct
tactaataaa ggagaagtat ttaatgaact tgttggaaaa 840cagcgcatat ctggaaattt
ggattctcca gaaggtggtt tcgatgccat catgcaagtt 900gcagtttgtg gatcactgat
tggctggagg aatgttacac ggctgctggt gttttccaca 960gatgccgggt ttcactttgc
tggagatggg aaacttggtg gcattgtttt accaaatgat 1020ggacaatgtc acctggaaaa
taatatgtac acaatgagcc attattatga ttatccttct 1080attgctcacc ttgtccagaa
actgagtgaa aataatattc agacaatttt tgcagttact 1140gaagaatttc agcctgttta
caaggagctg aaaaacttga tccctaagtc agcagtagga 1200acattatctg caaattctag
caatgtaatt cagttgatca ttgatgcata caattccctt 1260tcctcagaag tcattttgga
aaacggcaaa ttgtcagaag gagtaacaat aagttacaaa 1320tcttactgca agaacggggt
gaatggaaca ggggaaaatg gaagaaaatg ttccaatatt 1380tccattggag atgaggttca
atttgaaatt agcataactt caaataagtg tccaaaaaag 1440gattctgaca gctttaaaat
taggcctctg ggctttacgg aggaagtaga ggttattctt 1500cagtacatct gtgaatgtga
atgccaaagc gaaggcatcc ctgaaagtcc caagtgtcat 1560gaaggaaatg ggacatttga
gtgtggcgcg tgcaggtgca atgaagggcg tgttggtaga 1620cattgtgaat gcagcacaga
tgaagttaac agtgaagaca tggatgctta ctgcaggaaa 1680gaaaacagtt cagaaatctg
cagtaacaat ggagagtgcg tctgcggaca gtgtgtttgt 1740aggaagaggg ataatacaaa
tgaaatttat tctggcaaat tctgcgagtg tgataatttc 1800aactgtgata gatccaatgg
cttaatttgt ggaggaaatg gtgtttgcaa gtgtcgtgtg 1860tgtgagtgca accccaacta
cactggcagt gcatgtgact gttctttgga tactagtact 1920tgtgaagcca gcaacggaca
gatctgcaat ggccggggca tctgcgagtg tggtgtctgt 1980aagtgtacag atccgaagtt
tcaagggcaa acgtgtgaga tgtgtcagac ctgccttggt 2040gtctgtgctg agcataaaga
atgtgttcag tgcagagcct tcaataaagg agaaaagaaa 2100gacacatgca cacaggaatg
ttcctatttt aacattacca aggtagaaag tcgggacaaa 2160ttaccccagc cggtccaacc
tgatcctgtg tcccattgta aggagaagga tgttgacgac 2220tgttggttct attttacgta
ttcagtgaat gggaacaacg aggtcatggt tcatgttgtg 2280gagaatccag agtgtcccac
tggtccagac atcattccaa ttgtagctgg tgtggttgct 2340ggaattgttc ttattggcct
tgcattactg ctgatatgga agcttttaat gataattcat 2400gacagaaggg agtttgctaa
atttgaaaag gagaaaatga atgccaaatg ggacacgggt 2460gaaaatccta tttataagag
tgccgtaaca actgtggtca atccgaagta tgagggaaaa 2520tgagtactgc ccgtgcaaat
cccacaacac tgaatgcaaa gtagcaattt ccatagtcac 2580agttaggtag ctttagggca
atattgccat ggttttactc atgtgcaggt tttgaaaatg 2640tacaatatgt ataattttta
aaatgtttta ttattttgaa aataatgttg taattcatgc 2700cagggactga caaaagactt
gagacaggat ggttattctt gtcagctaag gtcacattgt 2760gcctttttga ccttttcttc
ctggactatt gaaatcaagc ttattggatt aagtgatatt 2820tctatagcga ttgaaagggc
aatagttaaa gtaatgagca tgatgagagt ttctgttaat 2880catgtattaa aactgatttt
tagctttaca aatatgtcag tttgcagtta tgcagaatcc 2940aaagtaaatg tcctgctagc
tagttaagga ttgttttaaa tctgttattt tgctatttgc 3000ctgttagaca tgactgatga
catatctgaa agacaagtat gttgagagtt gctggtgtaa 3060aatacgtttg aaatagttga
tctacaaagg ccatgggaaa aattcagaga gttaggaagg 3120aaaaaccaat agctttaaaa
cctgtgtgcc attttaagag ttacttaatg tttggtaact 3180tttatgcctt cactttacaa
attcaagcct tagataaaag aaccgagcaa ttttctgcta 3240aaaagtcctt gatttagcac
tatttacata caggccatac tttacaaagt atttgctgaa 3300tggggacctt ttgagttgaa
tttattttat tatttttatt ttgtttaatg tctggtgctt 3360tctatcacct cttctaatct
tttaatgtat ttgtttgcaa ttttggggta agactttttt 3420atgagtactt tttctttgaa
gttttagcgg tcaatttgcc tttttaatga acatgtgaag 3480ttatactgtg gctatgcaac
agctctcacc tacgcgagtc ttactttgag ttagtgccat 3540aacagaccac tgtatgttta
cttctcacca tttgagttgc ccatcttgtt tcacactagt 3600cacattcttg ttttaagtgc
ctttagtttt aacagttcac tttttacagt gctatttact 3660gaagttattt attaaatatg
cctaaaatac ttaaatcgga 3700276407DNAHomo sapiens
27agtctgcggg cctccggggc tgcggcgagg ccggagcgtc gcggcggaga ggacgagacc
60gggacaagac cagggcagga gggagccggc cagccgcgag aaccccgcac gcccggcaag
120atgctgtcct tgcggctgca gacgggcccc gagaaggccg agctccagga gctcaacgcc
180cggctctatg actacgtgtg tcgggtgcgg gagctggagc gcgaaaacct actcctggag
240gaggagctgc gcggccggcg cgggcgagag ggcctgtggg ccgaggggca ggcccgctgc
300gccgaggagg cgcgcagctt gcggcagcag ctggacgagc tgagctgggc cactgcgctg
360gcggagggcg agcgggacgc tctgcggcgc gagctgcggg agctgcagcg cctggatgcg
420gaggagcgcg ccgcccgcgg ccgcctggac gccgagctgg gtgcgcagca gcgcgagctg
480caggaggcgc tgggcgcgcg cgccgccctc gaggcgctgc tgggccgact gcaggccgag
540cgccgaggcc tcgacgcggc ccacgaacgc gacgtgaggg agctgcgcgc gcgcgccgcc
600agccttacca tgcatttccg cgcccgcgcc accggccccg ccgcgccgcc gccacgcctg
660cgggaggtgc acgacagcta cgcactgctg gtggccgagt cgtggcggga gacggtgcag
720ctgtacgagg acgaggtgcg cgagctggag gaggcgctgc ggcgcggcca ggagagcaga
780ctccaggcgg aggaagagac gcggctgtgc gcgcaggagg cagaggcgct gcggcgcgag
840gcgctcgggt tggagcagct gcgcgcgcgg ctggaggacg cgctgctgcg gatgcgcgag
900gagtacggga tacaggccga ggagcggcag agagtgattg actgcctgga ggatgagaag
960gcaaccctca ccttggccat ggctgactgg ctgcgggact atcaggacct cctgcaggtg
1020aagaccggcc tcagtttgga ggtggcgacc taccgggcct tattggaagg agaaagtaat
1080ccagagatag tgatctgggc tgagcacgtt gaaaacatgc cgtcagaatt cagaaacaaa
1140tcctatcact ataccgactc actactacag agggaaaatg aaaggaatct attttcaagg
1200cagaaagcac ctttggcaag tttcaatcac agctcggcac tgtattctaa cctgtcaggg
1260caccgtggat ctcagacggg cacatctatt ggaggtgatg ccagaagagg cttcttgggc
1320tcgggatatt cttcctcggc cactacccag caggaaaact catacggaaa agccgtcagc
1380agtcaaacca acgtcagaac tttctctcca acctatggcc ttttaagaaa tactgaggct
1440caagtgaaaa cattccctga cagaccaaaa gccggagata caagggaggt ccccgtttac
1500ataggtgaag attccacaat tgcccgcgag tcgtaccggg atcgccgaga caaggtggca
1560gcaggtgctt cggaaagcac acggtcaaat gagaggaccg tcattctggg aaagaaaaca
1620gaagtgaaag ccacgaggga gcaagaaaga aacagaccag aaaccatccg aacaaagcca
1680gaagagaaaa tgttcgattc taaagagaag gcttccgagg agagaaacct aagatgggaa
1740gaattgacaa agttagataa ggaagcgaga cagagagaaa gccagcagat gaaggagaag
1800gctaaggaga aggactcact gaaggagaag agtgtgcgag agagagaggt gccgattagt
1860ctagaagtat cccaggacag aagagcagag gtgtccccga aaggtttgca gacgcctgtg
1920aaggatgctg gtggtgggac cggtagagag gcagaagcaa gagagctacg gttcaggttg
1980ggcaccagtg atgccactgg ttctctgcaa ggcgattcca tgacagaaac cgtagcagaa
2040aacatcgtta ccagtatcct gaagcagttc actcagtctc cagagacaga agcatctgct
2100gattcttttc cagacacaaa agtcacttac gtggacagga aagagcttcc tggggaaagg
2160aaaacaaaga ctgaaatact tgtggagtct aaactgactg aggatgttga tgtttccgat
2220gaagctggcc tggactacct tttaagcaag gatattaagg aagtggggct gaaaggcaag
2280tcagccgagc agatgatagg agacatcatc aacctcggcc tgaaagggag ggaggggaga
2340gcaaaggtcg tcaacgtgga gatcgtggag gagcccgtga gttatgtcag cggggagaag
2400ccggaggagt tttccgtccc attcaaagtg gaggaggtcg aagatgtgtc gccaggcccc
2460tgggggttgg ttaaggagga ggaaggttat ggagaaagcg atgtcacatt ctcagttaat
2520cagcatcgaa ggaccaagca gcctcaggag aacacgactc acgtggaaga agtgacagag
2580gcaggtgatt cagagggcga gcagagttat tttgtgtcca ctccagatga acaccccggg
2640gggcacgaca gagatgacgg ctcggtgtac gggcagatcc acatcgagga ggaatccacc
2700atcaggtact cttggcagga tgaaatcgtg caggggactc gaaggaggac acagaaggac
2760ggtgcagtgg gcgagaaggt tgtgaagccc ttggatgtcc cagcgccctc tctggagggg
2820gacctgggtt ccactcactg gaaagaacaa gctagaagcg gtgaatttca tgccgaaccc
2880acagtcattg aaaaagaaat taaaataccc cacgaattcc acacctccat gaagggcatc
2940tcctccaagg agccccggca gcagctggtg gaagtcatcg ggcagctgga ggaaaccctt
3000cccgagcgca tgagggagga gctgtccgcc ctcaccagag aggggcaggg tgggccgggg
3060agcgtttccg tggatgtcaa gaaggtccag ggtgctggtg gcagttccgt gaccctggtt
3120gctgaagtca acgtctcaca aactgtggat gccgatcggt tagacctgga ggaggtgagc
3180aaagatgagg ccagtgagat ggagaaggct gtggagtcgg tggttcggga gagcctgagc
3240aggcaacgca gcccagcgcc tggcagccca gatgaggaag gtggagcgga ggccccggct
3300gctggcattc gctttaggcg ttgggccacc cgggagctgt acatcccttc aggcgagagc
3360gaggttgctg gtggggcctc tcacagctcg ggacagcgca ctccccaggg cccagtgtcg
3420gccactgtgg aggtcagcag ccccacaggc tttgcccagt cacaggtgct ggaggatgtg
3480agccaggctg caaggcacat aaaactcggc ccctctgaag tctggaggac tgagcgaatg
3540tcatatgaag gacccactgc agaagtggtg gagatggatg tgagtaacgt agaggcgatc
3600cgcagccgga cacaggaagc gggagctctc ggtgtgtctg accgtggttc ctggagagac
3660gcggacagta ggaatgacca ggcagttggt gtgagcttta aggcctctgc tggggaagga
3720gaccaggccc acagagaaca gggcaaggag caggccatgt ttgataagaa ggtgcagctc
3780cagagaatgg tagaccaaag gtcggtgatt tcagatgaaa agaaagttgc cctcctctat
3840ctagacaatg aggaggagga gaatgatggg cattggtttt aataagcaga aacattttgt
3900tttaatggca gcctgttggc gacgtgccaa catccaaagg ccttaactta ttttaagagg
3960ccgagggagt ctatgaaaat ctcccctttt ttactttttt aaagagtact cccggcatgg
4020tcaatttcct ttatagttaa tccgtaaagg tttccagtta attcatgcct taaaaggcac
4080tgcaatttta tttttgagtt gggactttta caaaacactt ttttccctgg agtcttctct
4140ccacttctgg agatgaattt ctatgttttg cacctggtca cagacatggc ttgcatctgt
4200ttgaaactac aattaattat agatgtcaaa acattaacca gattaaagta atatatttaa
4260gagtaaattt tgcttgcatg tgctaatatg aaataacaga ctaacatttt aggggaaaaa
4320taaatacaat ttagactcta aaaagtcttt tcaaaaagaa atgggaaata ggcagactgt
4380ttatgttaaa aaaattcttg ctaaatgatt tcatctttag gaaaaaatta cttgccatat
4440agagctaaat tcatcttaag acttgaatga attgctttct atgtacagaa ctttaaacaa
4500tatagtattt atggcgagga cagctgtagt ctgttgtgat atttcacatt ctatttgcac
4560aggttccctg gcactggtag ggtagatgat tattgggaat cgcttacagt accatttcat
4620tttttggcac taggtcatta agtagcacac agtctgaatg cccttttctg gagtggccag
4680ttcctatcag actgtgcaga cttgcgcttc tctgcacctt atcccttagc acccaaacat
4740ttaatttcac tggtgggagg tagaccttga agacaatgaa gagaatgccg atactcagac
4800tgcagctgga ccggcaagct ggctgtgtac aggaaaattg gaagcacaca gtggactgtg
4860cctcttaaag atgcctttcc caaccctcca ttcatgggat gcaggtcttt ctgagctcaa
4920gggtgaaaga tgaatacaat aacaaccatg aacccacctc acggaagctt tttttgcact
4980ttgaacagaa gtcattgcag ttggggtgtt ttgtccaggg aaacagttta ttaaatagaa
5040ggatgttttg gggaaggaac tggatatctc tcctgcagcc cagcaccgag atacccagga
5100cgggcctggg gggcgagaaa ggcccccatg ctcatgggcc gcggagtgtg gacctgtaga
5160taggcaccac cgagtttaag atactgggat gagcatgctt cattggattc attttatttt
5220acacgtcagt attgttttaa agtttctgtc tgtaaagtgt agcatcatat ataaaaagag
5280tttcgctagc agcgcatttt ttttagttca ggctagcttc tttcacataa tgctgtctca
5340gctgtatttc cagtaacaca gcatcatcac actgactgtg gcgcactggg gaataacagt
5400ctgagctagc accaccctca gccaggctac aacgacagca ctggagggtc ttccctctca
5460gattcacctg gaggccctca gacccccagg gtgcacgtct ccccaggtcc tgggagtggc
5520taccgcaggt agtttctgga gagcacgttt tcttcattga taagtggagg agaaatgcag
5580cacagctttc aagatactat tttaaaaaca ccatgaatca gatagggaaa gaaagttgat
5640tggaatggca agtttaaacc tttgttgtcc atctgccaaa tgaactagtg attgtcagac
5700tggtatggag gtgactgctt tgtaaggttt tgtcgtttct aatacagaca gagatgtgct
5760gattttgttt tagctgtaac aggtaatggt ttttggatag atgattgact ggtgagaatt
5820tggtcaaggt gacagcctcc tgtctgatga caggacagac tggtggtgag gagtctaagt
5880gggctcagtt tgatgtcagt gtctgggctc atgacttgta aatggaagct gatgtgaaca
5940ggtaattaat attatgaccc acttctattt actttgggaa atatcttgga tcttaattat
6000catctgcaag tttcaagaag tattctgcca aaagtattta caagtatgga ctcatgagct
6060attgttggtt gctaaatgtg aatcacgcgg gagtgagtgt gcccttcaca ctgtgacatt
6120gtgacattgt gacaagctcc atgtccttta aaatcagtca ctctgcacac aagagaaatc
6180aacttcgtgg ttggatgggg ccggaacaca accagtcttt ttgtatttat tgttactgag
6240acaaaacagt actcactgag tgtttttcag tttcctactg gtggttttga tattgtttgt
6300ttaagatgta tatttagaat gacatcatct aagaagctga ttttgctaaa ctcctgttcc
6360ctacaatggg aaatgtcaca agaatgtgca aaaataaaaa tctgagg
6407281295DNAHomo sapiens 28cacgtcagcc ggggctagaa aaggcggcgg ggctgggccc
agcgaggtga cagcctcgct 60tggacgcaga gcccggcccg acgccgccat gagcgccgcg
ctcttcagcc tggacggccc 120ggcgcgcggc gcgccctggc ctgcggagcc tgcgcccttc
tacgaaccgg gccgggcggg 180caagccgggc cgcggggccg agccaggggc cctaggcgag
ccaggcgccg ccgcccccgc 240catgtacgac gacgagagcg ccatcgactt cagcgcctac
atcgactcca tggccgccgt 300gcccaccctg gagctgtgcc acgacgagct cttcgccgac
ctcttcaaca gcaatcacaa 360ggcgggcggc gcggggcccc tggagcttct tcccggcggc
cccgcgcgcc ccttgggccc 420gggccctgcc gctccccgcc tgctcaagcg cgagcccgac
tggggcgacg gcgacgcgcc 480cggctcgctg ttgcccgcgc aggtggccgc gtgcgcacag
accgtggtga gcttggcggc 540cgcagggcag cccaccccgc ccacgtcgcc ggagccgccg
cgcagcagcc ccaggcagac 600ccccgcgccc ggccccgccc gggagaagag cgccggcaag
aggggcccgg accgcggcag 660ccccgagtac cggcagcggc gcgagcgcaa caacatcgcc
gtgcgcaaga gccgcgacaa 720ggccaagcgg cgcaaccagg agatgcagca gaagttggtg
gagctgtcgg ctgagaacga 780gaagctgcac cagcgcgtgg agcagctcac gcgggacctg
gccggcctcc ggcagttctt 840caagcagctg cccagcccgc ccttcctgcc ggccgccggg
acagcagact gccggtaacg 900cgcggccggg gcgggagaga ctcagcaacg acccatacct
cagacccgac ggcccggagc 960ggagcgcgcc ctgccctggc gcagccagag ccgccgggtg
cccgctgcag tttcttggga 1020cataggagcg caaagaagct acagcctgga cttaccacca
ctaaactgcg agagaagcta 1080aacgtgttta ttttccctta aattattttt gtaatggtag
ctttttctac atcttactcc 1140tgttgatgca gctaaggtac atttgtaaaa agaaaaaaaa
ccagactttt cagacaaacc 1200ctttgtattg tagataagag gaaaagactg agcatgctca
cttttttata ttaattttta 1260cagtatttgt aagaataaag cagcatttga aatcg
1295291526DNAHomo sapiens 29ggccactgct caccatgcac
ataacccagc tcaaccggga gtgcctgctg cacctcttct 60ccttcctaga caaggacagc
aggaagagcc ttgccaggac ctgctcccag ctccacgacg 120tgtttgagga ccccgcactc
tggtccctgc tgcacttccg ttccctcact gaactccaga 180aggacaactt cctcctgggc
ccggcactcc gcagcctctc catctgctgg cactccagcc 240gcgtgcaggt gtgcagcatt
gaggactggc tcaagagtgc cttccagaga agcatctgca 300gccggcacga gagcctggtc
aatgatttcc tcctccgggt gtgcgacagg ctttctgctg 360tgcgctcccc acggaggcgg
gaggcgcctg caccgtcctc ggggactccg atcgccgttg 420gaccgaaatc acctcggtgg
ggaggacctg accactcgga gttcgccgac ttgcgctcgg 480gggtgacggg ggccagggct
gccgcgcgca ggggtctggg gagcctccgg gcggagcgac 540ccagcgagac cccgccggct
cccggagtgt cctggggacc gccacctcca ggagccccgg 600tggtgatctc ggtgaagcag
gaggagggga agcaggggcg cacgggcaga aggagccacc 660gagccgctcc tccttgcggt
tttgcccgca cgcgcgtctg cccgcccacc tttcctgggg 720cggatgcgtt cccgcagtga
ccgcactcgc gattgtagaa aattcgctcc caattgttga 780atgcttacat aaactgcata
gttacccata ttttctactg ttctcataag ggaaaaagaa 840atgatttgga agaaaaaaaa
agcctcaaac taaacaaaaa caaaaccagc agccctggct 900aagcttttga taatggaacc
attataagct gacaagctag atttttcacc tgttggattt 960gctggacatg agcacaagcc
taggcttcca atttgaattt cagggctgta gtcactgatg 1020tgtcacattt agtgaaggct
ctcggccccc cggcagccgc ctggtcctgg gtcactgggg 1080agaattctgg gtgggtggaa
gaggcgagga gcactgtgat gggcgctgtt cacactccgg 1140ctccacccag cgccagctgc
gacaaagccc tttacccctc ccaggccagg tgtttcttct 1200gtgtgaaaga ggagcttgat
ccctactcac aggttcctta attcattgct caaagaggga 1260aacagcaagg aaaaagacaa
attctgttct catgaagctt accttctagt gggggaagag 1320acaacaggta actggtatat
aagagggtag aagggccagg tgcggtggct caatgcctgt 1380aatcccagca ctttaggagg
ccaagatggg cgattcacga ggtcaggaga tcgagaccat 1440cctggctaac aaggtgaaac
ccccgtctgt actaaaaaaa aaaaaaaaaa aaaaaaaaaa 1500aaaaaaaaaa aaaaaaaaaa
aaaaaa 1526301353DNAHomo sapiens
30atgcgggaga tcgtgcacat ccaggccggc cagtgcggca accagatcgg ggccaagttc
60tgggaagtca tcagtgatga gcatggcatc gaccccagcg gcaactacgt gggcgactcg
120gacttgcagc tggagcggat cagcgtctac tacaacgagg cctcttctca caagtacgtg
180cctcgagcca ttctggtgga cctggaaccc ggaaccatgg acagtgtccg ctcaggggcc
240tttggacatc tcttcaggcc tgacaatttc atctttggtc agagtggggc cggcaacaac
300tgggccaagg gtcactacac ggagggggcg gagctggtgg attcggtcct ggatgtggtg
360cggaaggagt gtgaaaactg cgactgcctg cagggcttcc agctgaccca ctcgctgggg
420ggcggcacgg gctccggcat gggcacgttg ctcatcagca aggtgcgtga ggagtatccc
480gaccgcatca tgaacacctt cagcgtcgtg ccctcaccca aggtgtcaga cacggtggtg
540gagccctaca acgccacgct gtccatccac cagctggtgg agaacacgga tgagacctac
600tgcatcgaca acgaggcgct ctacgacatc tgcttccgca ccctcaagct ggccacgccc
660acctacgggg acctcaacca cctggtatcg gccaccatga gcggagtcac cacctccttg
720cgcttcccgg gccagctcaa cgctgacctg cgcaagctgg ccgtcaacat ggtgcccttc
780ccgcgcctgc acttcttcat gcccggcttc gcccccctca cagcccgggg cagccagcag
840taccgggccc tgaccgtgcc cgagctcacc cagcagatgt tcgatgccaa gaacatgatg
900gccgcctgcg acccgcgcca cggccgctac ctgacggtgg ccaccgtgtt ccggggccgc
960atgtccatga aggaggtgga cgagcagatg ctggccatcc agagcaagaa cagcagctac
1020ttcgtggagt ggatccccaa caacgtgaag gtggccgtgt gtgacatccc gccccgcggc
1080ctcaagatgt cctccacctt catcgggaac agcacggcca tccaggagct gttcaagcgc
1140atctccgagc agttcacggc catgttccgg cgcaaggcct tcctgcactg gtacacgggc
1200gagggcatgg acgagatgga gttcaccgag gccgagagca acatgaacga cctggtgtcc
1260gagtaccagc agtaccagga cgccacggcc gaggaagagg gcgagatgta cgaagacgac
1320gaggaggagt cggaggccca gggccccaag tga
1353312853DNAHomo sapiens 31gctctgctcc aggcatctgc cacaatgtgg gtgcttacac
ctgctgcttt tgctgggaag 60ctcttgagtg tgttcaggca acctctgagc tctctgtgga
ggagcctggt cccgctgttc 120tgctggctga gggcaacctt ctggctgcta gctaccaaga
ggagaaagca gcagctggtc 180ctgagagggc cagatgagac caaagaggag gaagaggacc
ctcctctgcc caccacccca 240accagcgtca actatcactt cactcgccag tgcaactaca
aatgcggctt ctgtttccac 300acagccaaaa catcctttgt gctgcccctt gaggaagcaa
agagaggatt gcttttgctt 360aaggaagctg gtatggagaa gatcaacttt tcaggtggag
agccatttct tcaagaccgg 420ggagaatacc tgggcaagtt ggtgaggttc tgcaaagtag
agttgcggct gcccagcgtg 480agcatcgtga gcaatggaag cctgatccgg gagaggtggt
tccagaatta tggtgagtat 540ttggacattc tcgctatctc ctgtgacagc tttgacgagg
aagtcaatgt ccttattggc 600cgtggccaag gaaagaagaa ccatgtggaa aaccttcaaa
agctgaggag gtggtgtagg 660gattatagag tcgctttcaa gataaattct gtcattaatc
gtttcaacgt ggaagaggac 720atgacggaac agatcaaagc actaaaccct gtccgctgga
aagtgttcca gtgcctctta 780attgagggtg agaattgtgg agaagatgct ctaagagaag
cagaaagatt tgttattggt 840gatgaagaat ttgaaagatt cttggagcgc cacaaagaag
tgtcctgctt ggtgcctgaa 900tctaaccaga agatgaaaga ctcctacctt attctggatg
aatatatgcg ctttctgaac 960tgtagaaagg gacggaagga cccttccaag tccatcctgg
atgttggtgt agaagaagct 1020ataaaattca gtggatttga tgaaaagatg tttctgaagc
gaggaggaaa atacatatgg 1080agtaaggctg atctgaagct ggattggtag agcggaaagt
ggaacgagac ttcaacacac 1140cagtgggaaa actcctagag taactgccat tgtctgcaat
actatcccgt tggtatttcc 1200cagtggctga aaacctgatt ttctgctgca cgtggcatct
gattacctgt ggtcactgaa 1260cacacgaata acttggatag caaatcctga gacaatggaa
aaccattaac tttacttcat 1320tggcttataa ccttgttgtt attgaaacag cacttctgtt
tttgagtttg ttttagctaa 1380aaagaaggaa tacacacagg aataatgacc ccaaaaatgc
ttagataagg cccctataca 1440caggacctga catttagctc aatgatgcgt ttgtaagaaa
taagctctag tgatatctgt 1500gggggcaaaa tttaatttgg atttgatttt ttaaaacaat
gtttactgcg atttctatat 1560ttccattttg aaactatttc ttgttccagg tttgttcatt
tgacagagtc agtatttttt 1620gccaaatatc cagataacca gttttcacat ctgagacatt
acaaagtatc tgcctcaatt 1680atttctgctg gttataatgc tttttttttt ttgcctttat
gccattgcag tcttgtactt 1740tttactgtga tgtacagaaa tagtcaacag atgtttccaa
gaacatatga tatgataatc 1800ctaccaattt tcaagaagtc tctagaaaga gataacacat
ggaaagacgg cgtggtgcag 1860cccagcccac ggtgcctgtt ccatgaatgc tggctaccta
tgtgtgtggt acctgttgtg 1920tccctttctc ttcaaagatc cctgagcaaa acaaagatac
gctttccatt tgatgatgga 1980gttgacatgg aggcagtgct tgcattgctt tgttcgccta
tcatctggcc acatgaggct 2040gtcaagcaaa agaataggag tgtagttgag tagctggttg
gccctacatt tctgagaagt 2100gacgttacac tgggttggca taagatatcc taaaatcacg
ctggaacctt gggcaaggaa 2160gaatgtgagc aagagtagag agagtgcctg gatttcatgt
cagtgaagcc atgtcaccat 2220atcatatttt tgaatgaact ctgagtcagt tgaaataggg
taccatctag gtcagtttaa 2280gaagagtcag ctcagagaaa gcaagcataa gggaaaatgt
cacgtaaact agatcaggga 2340acaaaatcct ctccttgtgg aaatatccca tgcagtttgt
tgatacaact tagtatctta 2400ttgcctaaaa aaaaatttct tatcattgtt tcaaaaaagc
aaaatcatgg aaaatttttg 2460ttgtccaggc aaataaaagg tcattttaat ttaaaaaaaa
aaaaaaaaaa aaaaaaaaaa 2520aaaaggccaa ggaaaaaaaa tattcctact taaattttaa
gtctataatt caatttaaat 2580atgtgtgtgt ctcatccagg ataggatagg ttgtcttcta
ttttccattt tacctattta 2640ctttttttgt aagaaaagag aagaatgaat tctaaagatg
ttccccatgg gttttgattg 2700tgtctaagct atgatgacct tcatataatc agcataaaca
taaaacaaat tttttactta 2760acatgagtgc actttactaa tcctcatggc acagtggctc
acgcctgtaa tcccagcact 2820tggggaggac aatgtggggt ggatcacgag gtc
2853326853DNAHomo sapiens 32atacatagac ttccgttaaa
accagaatga ggaaaatact tttctctagc atcgtaggag 60gaagaaaaca aacacatcag
atattttcag cactaaaaga gatggttttc cccacatata 120tgtaaaagaa atttgcaaga
ctactggatt ttgatctcat ggttgcagtg ggtgaatagg 180tggccttttg tgatctccta
catcaccctg gaagtgagac ttcttcggtt tcttctagag 240tcagtttggt atcagaatgg
caaagcaact taaccttcca gaaaatacag atgattggac 300aaaagaggat gtaaatcagt
ggttagaaag tcataagatt gaccaaaaac acagggaaat 360tttgactgaa caagacgtga
atggagcagt cttgaagtgg ttaaaaaaag aacatcttgt 420tgatatgggc atcacacatg
gaccagctat tcaaatagaa gaactattca aagaattgcg 480gaaaacagcc attgaagatt
cgattcagac atctaagatg ggaaagccca gtaaaaatgc 540tcctaaagac caaactgtgt
ctcaaaagga acgtagagaa acttcaaagc aaaaacaaaa 600gggtaaagag aacccagata
tggctaatcc gtctgcaatg agtacaactg ctaaaggttc 660taagtcacta aaagttgagc
tcatagaaga taaaatagat tatacaaagg aaaggcaacc 720atccatagac ctgacatgtg
tatcatatcc atttgatgaa ttcagtaatc catatcgtta 780caagttggat tttagtctac
agcctgaaac aggaccaggc aatctcattg atccgataca 840tgaattcaaa gccttcacaa
atacagcaac agccacagaa gaggatgtca agatgaaatt 900tagcaatgag gttttccgat
ttgcttcagc ttgtatgaat tcacgtacca atggcactat 960tcattttgga gtcaaagaca
aaccccatgg gaaaattgtt ggcatcaaag tcaccaatga 1020taccaaggaa gccctcatta
accatttcaa tctgatgata aacaagtatt ttgaagacca 1080tcaagtccaa caagcaaaga
agtgcattcg agagccaaga tttgtggaag ttttactgcc 1140aaatagtact ctatctgaca
gatttgttat tgaagtggac attattccac agttctctga 1200atgccaatat gattatttcc
agattaaaat gcaaaattac aacaacaaaa tatgggaaca 1260aagtaaaaaa ttctcactat
ttgtgcgaga tgggaccagc tctaaggaca ttacgaaaaa 1320taaagttgat ttcagagcat
ttaaagcaga ttttaaaaca ctggcagagt ccagaaaagc 1380agcagaagaa aaattcagag
caaaaacaaa taaaaaagaa agagagggac caaagttggt 1440taaattattg acaggaaatc
aagatttgtt agataattca tactatgaac agtacattct 1500tgtaacaaat aaatgccacc
cagatcaaac aaaacactta gatttcctga aggaaattaa 1560atggtttgct gtattggagt
ttgatcctga gtctaacatc aatggagtgg tcaaagctta 1620caaagaaagc cgagtagcaa
accttcactt tccaagtgta tatgtagaac agaaaaccac 1680accaaatgag acgatttcta
ctctaaatct ttaccatcaa cccagctgga ttttctgcaa 1740tggcaggtta gaccttgaca
gtgaaaaata taaacccttt gatccaagtt cctggcaaag 1800agaaagagct tctgatgtca
ggaaactgat ttcatttctt acacatgaag acataatgcc 1860aagagggaag tttttggtgg
tatttctatt actgtcctct gtggatgacc caagagatcc 1920cctcattgag actttctgtg
ctttctacca ggatctcaaa ggaatggaaa atatactgtg 1980tatttgtgtg cacccacaca
tatttcaggg atggaaagat ctacttgaag caagattaat 2040aaaacaccaa gatgaaattt
caagccaatg tatttctgct ttaagccttg aagagatcaa 2100tggcactatt cttaaactaa
aatctgtgac tcaatcttca aaaaggcttt tgccatctat 2160tggtttatcg actgtccttc
tgaaaaagga agaagatatc atgactgctc tggaaattat 2220ctgtgaaaat gaatgtgagg
gtacactgtt agagaaggac aaaaataaat tccttgaatt 2280caaggcatca aaagaggaag
acttctatcg aggtggcaaa gtgtcatggt ggaacttcta 2340cttctcttct gaaagttatt
cttcaccttt tgtcaaaagg gataaatatg aaagacttga 2400agcaatgatt caaaactgtg
cagattcttc taaaccaaca agtaccaaaa ttattcatct 2460gtatcatcat ccaggctgtg
ggggaactac cttggctatg cacattctct gggaactaag 2520gaagaaattc agatgtgctg
tgctgaaaaa caagacagtg gatttttctg aaattggaga 2580acaggtaacc agtttaatca
cctatggggc aatgaaccgt caggaatacg tacctgtact 2640actccttgtt gatgattttg
aagaacaaga taatgtctat cttctgcagt actctattca 2700aacagctata gctaaaaagt
acattcgata tgaaaaacct ctggtgatta tcctaaattg 2760tatgagatca caaaatcctg
aaaaaagtgc aaggatccca gacagtattg ccgtaataca 2820gcaactctct cccaaagaac
agagagcttt tgagcttaaa ttgaaagaaa tcaaagaaca 2880gcataaaaac tttgaggatt
tttattcctt tatgatcatg aaaaccaatt ttaataaaga 2940atacatagaa aatgtggtcc
ggaatatcct gaaagggcag aatattttca ccaaggaagc 3000aaagctcttt tcttttctgg
ctcttcttaa ttcatatgtg cctgatacca ccatttcact 3060atcacagtgt gaaaaattct
taggaattgg aaacaagaag gctttctggg ggacagaaaa 3120atttgaagac aagatgggca
cctactctac aattctgata aaaacagagg tcatcgaatg 3180tgggaactac tgtggagtac
gcatcattca ctctttgatt gcagagttct cactggaaga 3240attgaagaaa agctatcacc
tgaataaaag tcaaattatg ttggatatgc taactgagaa 3300tttgttcttc gatactggta
tgggaaaaag taaatttttg caagatatgc acacactcct 3360actcacaaga caccgcgatg
aacatgaagg tgaaacagga aattggtttt ccccatttat 3420tgaagcatta cataaagatg
aaggaaatga agcagttgaa gctgtattgc ttgaaagtat 3480ccatcggttc aacccaaatg
cattcatttg ccaagcgttg gcaagacatt tctacattaa 3540aaagaaggac tttggcaatg
ctctaaactg ggcaaaacaa gcaaaaatca tagaacctga 3600caattcttat atctcagata
cactgggtca agtctacaaa agtaaaataa gatggtggat 3660agaggaaaac ggaggaaacg
ggaacatttc agttgatgat ctaattgctc ttttggattt 3720agcagaacat gcctcaagtg
cattcaaaga atctcaacag caaagtgaag atagagagta 3780tgaagtgaag gaaagattgt
atccgaagtc aaaaaggcgg tatgatactt acaatatagc 3840tggttatcaa ggagagatag
aagttgggct ttacacaatc caaattctcc agctcattcc 3900tttttttgat aataaaaatg
agctatctaa aagatatatg gtcaattttg tatcaggaag 3960tagtgatatt ccaggggatc
caaacaatga atataaatta gccctcaaaa actatattcc 4020ttatttaact aaattgaaat
tttctttgaa aaagtccttt gatttttttg atgaatactt 4080tgtcctgcta aaacccagga
acaatattaa gcaaaatgaa gaggccaaaa ctcggagaaa 4140ggtggctgga tattttaaga
aatatgtaga tatattttgt ctcttagaag aatcacaaaa 4200caacacaggt cttggatcaa
agttcagtga gccacttcaa gtagagagat gcaggagaaa 4260cctagtagct ttaaaagcag
acaagttttc tgggctcttg gaatatctta tcaaaagtca 4320agaggatgct ataagcacta
tgaaatgtat agtgaacgaa tatacttttc tcttagaaca 4380atgcactgtc aaaatccagt
caaaagaaaa gctaaatttc atcttggcca acattattct 4440ctcctgtatc caacctacct
ccagattagt aaagccagtt gaaaaactaa aagatcagct 4500tcgagaagtc ttgcaaccaa
taggactgac ttatcagttt tcagaaccgt attttctagc 4560ttccctctta ttctggccag
aaaatcaaca actagatcaa cattctgaac aaatgaaaga 4620gtatgctcaa gcactaaaaa
attctttcaa ggggcaatat aaacatatgc atcgtacaaa 4680gcaaccaatt gcatatttct
ttcttggaaa aggtaaaaga ctggaaagac ttgttcacaa 4740aggaaaaatt gaccagtgct
ttaagaagac accagatatt aattccttgt ggcagagtgg 4800agatgtgtgg aaggaggaaa
aagtccaaga acttttgctt cgtttacaag gtcgagctga 4860aaacaattgt ttatatatag
aatatggaat caatgaaaaa atcacaatac ccatcactcc 4920cgctttttta ggtcaactta
gaagtggcag aagcatagag aaggtgtctt tttacctggg 4980attttccatt ggaggcccac
ttgcttatga cattgaaatt gtttaagagc ctgatattct 5040tcctccaaga atttgatctc
agtacccatt taattttttt ggactcaaga tctatgcttt 5100aaaccggcaa ggttatagat
acagcctcta gctcttcaga tctgtacatg cagtatttaa 5160tttcctctta aacatgttat
gagttctaca aagacaatag tgaaaaagga aggagtgaga 5220tatatgaaaa gtagcaaata
tgttccttgg tttggttaac atcattgatg acaaaataat 5280aaggagctat gactggagtc
aggagaagtt agtgtaataa gctggctaca cagaacccca 5340ctacttacca ggcatggatt
gaagaagatt gtctactcaa atggcattta gacattagaa 5400tgtctgggaa aatatttctc
aaagacagca aaaacctctc aaactgagga gcaacattta 5460ttcttactaa gcagatcatc
aatgtatcat gtgcttggca ctcaaggatc ttccaaaaca 5520gaggaccaac cagtcttctg
aaggtcatgc ccacagaagt catcagacct taccaaagta 5580ggttggagaa ttagattgcc
ttttcatgca gtgagattca gttaagcaaa aatgaaattt 5640gtctctatag ctaattagct
tatcaactcc cctccaaaca aacaattaaa aaaaaaacat 5700acagacactc aaattccaca
agctaatgaa caaaagggac tcttgtgaga agactaatga 5760gtccctcatc cagaagatgc
caatgtactg gcagattaac atacaaccta tgttttgaac 5820aaaaacaacc agcgatacgt
aatcaaaatg taattttccc ctaataaaat tatggatatg 5880ggcagtcatc aatggctgcc
aaaaccatta agtggaaagc tgattaaaaa acaaaaattt 5940ctaatggatt tatcaaactg
tcccaaatcc tgataaatat taacatcaca gaggaagacc 6000agacattatg ggcctggaag
tactatagga gtgcacacat cacccgtgac atggtcttgc 6060caaataatta aacctgaatt
tgatcaggtc tctggatctt atttgcaatt caaaagaaat 6120tttaaaaaaa tcctactaac
accaccacaa atatgcaatc agcaatatcc agaaagggga 6180aattcacagg acaaaaacct
ggttttcttt tttggtttct tcaaccaaaa aagaaagaaa 6240ttgcaaagga ccaaaaaaat
gttggggaat ctatacatta taagggactt aacaactaaa 6300gggcaacata tagactttag
atcctaattt gagcaaaatc taaaatcaat tattaggcaa 6360tcagaaaaat ttgaacacag
actagatatt tgaggatatt aaggtactat attattgaag 6420attccatggt tatgtttttt
aaagagttca tgccttttag agatacatac taaagtattt 6480gtaaataaat gacatgatct
agaatcagta attttgtgtt tggggtgtgg gggtggtgaa 6540agggaagtag aaccgaaaca
agattagtcc tgagttaaca atggctgcaa gctggataca 6600tggaattcag cacacttttc
tccctcttac tgattatgct tttgaaattt tctcttgtaa 6660aacatttaga aaacaaaaac
aaaaaaaatg tgatttgttt ctgtcttcaa aatctcatta 6720gaattttttc actggaggaa
gattttccct tgcttctgca taaaatttta actccataac 6780ttataagctc actctttatt
gttactttct aattgacaaa taaaaattgt atattaaaaa 6840aaaaaaaaaa aaa
685333672DNAHomo sapiens
33aacacatcca agcttaagac ggtgaggtca gcttcacatt ctcaggaact ctccttcttt
60gggtctggct gaagttgagg atctcttact ctctaggcca cggaattaac ccgagcaggc
120atggaggcct ctgctctcac ctcatcagca gtgaccagtg tggccaaagt ggtcagggtg
180gcctctggct ctgccgtagt tttgcccctg gccaggattg ctacagttgt gattggagga
240gttgtggcca tggcggctgt gcccatggtg ctcagtgcca tgggcttcac tgcggcggga
300atcgcctcgt cctccatagc agccaagatg atgtccgcgg cggccattgc caatgggggt
360ggagttgcct cgggcagcct tgtggctact ctgcagtcac tgggagcaac tggactctcc
420ggattgacca agttcatcct gggctccatt gggtctgcca ttgcggctgt cattgcgagg
480ttctactagc tccctgcccc tcgccctgca gagaagagaa ccatgccagg ggagaaggca
540cccagccatc ctgacccagc gaggagccaa ctatcccaaa tatacctggg gtgaaatata
600ccaaattctg catctccaga ggaaaataag aaataaagat gaattgttgc aactcttaaa
660aaaaaaaaaa aa
67234469PRTHomo sapiens 34Met Ser Gln Ala Tyr Ser Ser Ser Gln Arg Val Ser
Ser Tyr Arg Arg1 5 10
15Thr Phe Gly Gly Ala Pro Val Phe Pro Leu Gly Ser Pro Leu Ser Ser
20 25 30Pro Val Phe Pro Arg Ala Pro
Phe Gly Ser Lys Gly Ser Ser Ser Ser 35 40
45Val Thr Ser Arg Val Tyr Gln Val Ser Arg Thr Ser Gly Gly Ala
Gly 50 55 60Gly Leu Gly Ser Leu Arg
Ala Ser Arg Leu Gly Thr Thr Arg Thr Pro65 70
75 80Ser Ser Tyr Gly Ala Gly Glu Leu Leu Asp Phe
Ser Leu Ala Asp Ala 85 90
95Val Asn Gln Glu Phe Leu Thr Thr Arg Thr Asn Glu Lys Val Glu Leu
100 105 110Gln Glu Leu Asn Asp Arg
Ser Pro Ile Tyr Met Glu Lys Val Arg Phe 115 120
125Leu Glu Gln Gln Asn Ala Leu Ala Ala Glu Val Asn Arg Leu
Lys Gly 130 135 140Arg Glu Pro Thr Arg
Val Ala Glu Leu Tyr Glu Glu Glu Leu Arg Glu145 150
155 160Leu Arg Arg Gln Val Glu Val Leu Thr Asn
Gln Arg Ala Arg Val Asp 165 170
175Val Glu Arg Asp Asn Leu Leu Asp Asp Leu Gln Arg Leu Lys Ala Lys
180 185 190Leu Gln Glu Glu Ile
Gln Leu Lys Glu Glu Ala Glu Asn Asn Leu Ala 195
200 205Ala Phe Arg Ala Asp Val Asp Ala Ala Thr Leu Ala
Arg Ile Asp Leu 210 215 220Glu Arg Arg
Ile Glu Ser Leu Asn Glu Glu Ile Ala Phe Leu Lys Lys225
230 235 240Val His Glu Glu Glu Ile Arg
Glu Leu Gln Ala Gln Leu Gln Glu Gln 245
250 255Gln Val Gln Val Glu Met Asp Met Ser Lys Pro Asp
Leu Thr Ala Ala 260 265 270Leu
Arg Asp Ile Arg Ala Gln Tyr Glu Thr Ile Ala Ala Lys Asn Ile 275
280 285Ser Glu Ala Glu Glu Trp Tyr Lys Ser
Lys Val Ser Asp Leu Thr Gln 290 295
300Ala Ala Asn Lys Asn Asn Asp Ala Leu Arg Gln Ala Lys Gln Glu Met305
310 315 320Met Glu Tyr Arg
His Gln Ile Gln Ser Tyr Thr Cys Glu Ile Asp Ala 325
330 335Leu Lys Gly Thr Asn Asp Ser Leu Met Arg
Gln Met Arg Glu Leu Glu 340 345
350Asp Arg Phe Ala Ser Glu Ala Ser Gly Tyr Gln Asp Asn Ile Ala Arg
355 360 365Leu Glu Glu Glu Ile Arg His
Leu Lys Asp Glu Met Ala Arg His Leu 370 375
380Arg Glu Tyr Gln Asp Leu Leu Asn Val Lys Met Ala Leu Asp Val
Glu385 390 395 400Ile Ala
Thr Tyr Arg Lys Leu Leu Glu Gly Glu Glu Ser Arg Ile Asn
405 410 415Leu Pro Ile Gln Thr Tyr Ser
Ala Leu Asn Phe Arg Glu Thr Ser Pro 420 425
430Glu Gln Arg Gly Ser Glu Val His Thr Lys Lys Thr Val Met
Ile Lys 435 440 445Thr Ile Glu Thr
Arg Asp Gly Glu Val Val Ser Glu Ala Thr Gln Gln 450
455 460Gln His Glu Val Leu4653562PRTHomo sapiens 35Met
Ser Glu Arg Gln Gly Ala Gly Ala Thr Asn Gly Lys Asp Lys Thr1
5 10 15Ser Gly Glu Asn Asp Gly Gln
Lys Lys Val Gln Glu Glu Phe Asp Ile 20 25
30Asp Met Asp Ala Pro Glu Thr Glu Arg Ala Ala Val Ala Ile
Gln Ser 35 40 45Gln Phe Arg Lys
Phe Gln Lys Lys Lys Ala Gly Ser Gln Ser 50 55
6036328PRTHomo sapiens 36Met Leu Pro Arg Val Gly Cys Pro Ala Leu
Pro Leu Pro Pro Pro Pro1 5 10
15Leu Leu Pro Leu Leu Pro Leu Leu Leu Leu Leu Leu Gly Ala Ser Gly
20 25 30Gly Gly Gly Gly Ala Arg
Ala Glu Val Leu Phe Arg Cys Pro Pro Cys 35 40
45Thr Pro Glu Arg Leu Ala Ala Cys Gly Pro Pro Pro Val Ala
Pro Pro 50 55 60Ala Ala Val Ala Ala
Val Ala Gly Gly Ala Arg Met Pro Cys Ala Glu65 70
75 80Leu Val Arg Glu Pro Gly Cys Gly Cys Cys
Ser Val Cys Ala Arg Leu 85 90
95Glu Gly Glu Ala Cys Gly Val Tyr Thr Pro Arg Cys Gly Gln Gly Leu
100 105 110Arg Cys Tyr Pro His
Pro Gly Ser Glu Leu Pro Leu Gln Ala Leu Val 115
120 125Met Gly Glu Gly Thr Cys Glu Lys Arg Arg Asp Ala
Glu Tyr Gly Ala 130 135 140Ser Pro Glu
Gln Val Ala Asp Asn Gly Asp Asp His Ser Glu Gly Gly145
150 155 160Leu Val Glu Asn His Val Asp
Ser Thr Met Asn Met Leu Gly Gly Gly 165
170 175Gly Ser Ala Gly Arg Lys Pro Leu Lys Ser Gly Met
Lys Glu Leu Ala 180 185 190Val
Phe Arg Glu Lys Val Thr Glu Gln His Arg Gln Met Gly Lys Gly 195
200 205Gly Lys His His Leu Gly Leu Glu Glu
Pro Lys Lys Leu Arg Pro Pro 210 215
220Pro Ala Arg Thr Pro Cys Gln Gln Glu Leu Asp Gln Val Leu Glu Arg225
230 235 240Ile Ser Thr Met
Arg Leu Pro Asp Glu Arg Gly Pro Leu Glu His Leu 245
250 255Tyr Ser Leu His Ile Pro Asn Cys Asp Lys
His Gly Leu Tyr Asn Leu 260 265
270Lys Gln Cys Lys Met Ser Leu Asn Gly Gln Arg Gly Glu Cys Trp Cys
275 280 285Val Asn Pro Asn Thr Gly Lys
Leu Ile Gln Gly Ala Pro Thr Ile Arg 290 295
300Gly Asp Pro Glu Cys His Leu Phe Tyr Asn Glu Gln Gln Glu Ala
Cys305 310 315 320Gly Val
His Thr Gln Arg Met Gln 32537967PRTHomo sapiens 37Met Gln
Arg Ala Val Pro Glu Gly Phe Gly Arg Arg Lys Leu Gly Ser1 5
10 15Asp Met Gly Asn Ala Glu Arg Ala
Pro Gly Ser Arg Ser Phe Gly Pro 20 25
30Val Pro Thr Leu Leu Leu Leu Ala Ala Ala Leu Leu Ala Val Ser
Asp 35 40 45Ala Leu Gly Arg Pro
Ser Glu Glu Asp Glu Glu Leu Val Val Pro Glu 50 55
60Leu Glu Arg Ala Pro Gly His Gly Thr Thr Arg Leu Arg Leu
His Ala65 70 75 80Phe
Asp Gln Gln Leu Asp Leu Glu Leu Arg Pro Asp Ser Ser Phe Leu
85 90 95Ala Pro Gly Phe Thr Leu Gln
Asn Val Gly Arg Lys Ser Gly Ser Glu 100 105
110Thr Pro Leu Pro Glu Thr Asp Leu Ala His Cys Phe Tyr Ser
Gly Thr 115 120 125Val Asn Gly Asp
Pro Ser Ser Ala Ala Ala Leu Ser Leu Cys Glu Gly 130
135 140Val Arg Gly Ala Phe Tyr Leu Leu Gly Glu Ala Tyr
Phe Ile Gln Pro145 150 155
160Leu Pro Ala Ala Ser Glu Arg Leu Ala Thr Ala Ala Pro Gly Glu Lys
165 170 175Pro Pro Ala Pro Leu
Gln Phe His Leu Leu Arg Arg Asn Arg Gln Gly 180
185 190Asp Val Gly Gly Thr Cys Gly Val Val Asp Asp Glu
Pro Arg Pro Thr 195 200 205Gly Lys
Ala Glu Thr Glu Asp Glu Asp Glu Gly Thr Glu Gly Glu Asp 210
215 220Glu Gly Ala Gln Trp Ser Pro Gln Asp Pro Ala
Leu Gln Gly Val Gly225 230 235
240Gln Pro Thr Gly Thr Gly Ser Ile Arg Lys Lys Arg Phe Val Ser Ser
245 250 255His Arg Tyr Val
Glu Thr Met Leu Val Ala Asp Gln Ser Met Ala Glu 260
265 270Phe His Gly Ser Gly Leu Lys His Tyr Leu Leu
Thr Leu Phe Ser Val 275 280 285Ala
Ala Arg Leu Tyr Lys His Pro Ser Ile Arg Asn Ser Val Ser Leu 290
295 300Val Val Val Lys Ile Leu Val Ile His Asp
Glu Gln Lys Gly Pro Glu305 310 315
320Val Thr Ser Asn Ala Ala Leu Thr Leu Arg Asn Phe Cys Asn Trp
Gln 325 330 335Lys Gln His
Asn Pro Pro Ser Asp Arg Asp Ala Glu His Tyr Asp Thr 340
345 350Ala Ile Leu Phe Thr Arg Gln Asp Leu Cys
Gly Ser Gln Thr Cys Asp 355 360
365Thr Leu Gly Met Ala Asp Val Gly Thr Val Cys Asp Pro Ser Arg Ser 370
375 380Cys Ser Val Ile Glu Asp Asp Gly
Leu Gln Ala Ala Phe Thr Thr Ala385 390
395 400His Glu Leu Gly His Val Phe Asn Met Pro His Asp
Asp Ala Lys Gln 405 410
415Cys Ala Ser Leu Asn Gly Val Asn Gln Asp Ser His Met Met Ala Ser
420 425 430Met Leu Ser Asn Leu Asp
His Ser Gln Pro Trp Ser Pro Cys Ser Ala 435 440
445Tyr Met Ile Thr Ser Phe Leu Asp Asn Gly His Gly Glu Cys
Leu Met 450 455 460Asp Lys Pro Gln Asn
Pro Ile Gln Leu Pro Gly Asp Leu Pro Gly Thr465 470
475 480Ser Tyr Asp Ala Asn Arg Gln Cys Gln Phe
Thr Phe Gly Glu Asp Ser 485 490
495Lys His Cys Pro Asp Ala Ala Ser Thr Cys Ser Thr Leu Trp Cys Thr
500 505 510Gly Thr Ser Gly Gly
Val Leu Val Cys Gln Thr Lys His Phe Pro Trp 515
520 525Ala Asp Gly Thr Ser Cys Gly Glu Gly Lys Trp Cys
Ile Asn Gly Lys 530 535 540Cys Val Asn
Lys Thr Asp Arg Lys His Phe Asp Thr Pro Phe His Gly545
550 555 560Ser Trp Gly Met Trp Gly Pro
Trp Gly Asp Cys Ser Arg Thr Cys Gly 565
570 575Gly Gly Val Gln Tyr Thr Met Arg Glu Cys Asp Asn
Pro Val Pro Lys 580 585 590Asn
Gly Gly Lys Tyr Cys Glu Gly Lys Arg Val Arg Tyr Arg Ser Cys 595
600 605Asn Leu Glu Asp Cys Pro Asp Asn Asn
Gly Lys Thr Phe Arg Glu Glu 610 615
620Gln Cys Glu Ala His Asn Glu Phe Ser Lys Ala Ser Phe Gly Ser Gly625
630 635 640Pro Ala Val Glu
Trp Ile Pro Lys Tyr Ala Gly Val Ser Pro Lys Asp 645
650 655Arg Cys Lys Leu Ile Cys Gln Ala Lys Gly
Ile Gly Tyr Phe Phe Val 660 665
670Leu Gln Pro Lys Val Val Asp Gly Thr Pro Cys Ser Pro Asp Ser Thr
675 680 685Ser Val Cys Val Gln Gly Gln
Cys Val Lys Ala Gly Cys Asp Arg Ile 690 695
700Ile Asp Ser Lys Lys Lys Phe Asp Lys Cys Gly Val Cys Gly Gly
Asn705 710 715 720Gly Ser
Thr Cys Lys Lys Ile Ser Gly Ser Val Thr Ser Ala Lys Pro
725 730 735Gly Tyr His Asp Ile Ile Thr
Ile Pro Thr Gly Ala Thr Asn Ile Glu 740 745
750Val Lys Gln Arg Asn Gln Arg Gly Ser Arg Asn Asn Gly Ser
Phe Leu 755 760 765Ala Ile Lys Ala
Ala Asp Gly Thr Tyr Ile Leu Asn Gly Asp Tyr Thr 770
775 780Leu Ser Thr Leu Glu Gln Asp Ile Met Tyr Lys Gly
Val Val Leu Arg785 790 795
800Tyr Ser Gly Ser Ser Ala Ala Leu Glu Arg Ile Arg Ser Phe Ser Pro
805 810 815Leu Lys Glu Pro Leu
Thr Ile Gln Val Leu Thr Val Gly Asn Ala Leu 820
825 830Arg Pro Lys Ile Lys Tyr Thr Tyr Phe Val Lys Lys
Lys Lys Glu Ser 835 840 845Phe Asn
Ala Ile Pro Thr Phe Ser Ala Trp Val Ile Glu Glu Trp Gly 850
855 860Glu Cys Ser Lys Ser Cys Glu Leu Gly Trp Gln
Arg Arg Leu Val Glu865 870 875
880Cys Arg Asp Ile Asn Gly Gln Pro Ala Ser Glu Cys Ala Lys Glu Val
885 890 895Lys Pro Ala Ser
Thr Arg Pro Cys Ala Asp His Pro Cys Pro Gln Trp 900
905 910Gln Leu Gly Glu Trp Ser Ser Cys Ser Lys Thr
Cys Gly Lys Gly Tyr 915 920 925Lys
Lys Arg Ser Leu Lys Cys Leu Ser His Asp Gly Gly Val Leu Ser 930
935 940His Glu Ser Cys Asp Pro Leu Lys Lys Pro
Lys His Phe Ile Asp Phe945 950 955
960Cys Thr Met Ala Glu Cys Ser 96538620PRTHomo
sapiens 38Met Ser Tyr Tyr Gln Arg Pro Phe Ser Pro Ser Ala Tyr Ser Leu
Pro1 5 10 15Ala Ser Leu
Asn Ser Ser Ile Val Met Gln His Gly Thr Ser Leu Asp 20
25 30Ser Thr Asp Thr Tyr Pro Gln His Ala Gln
Ser Leu Asp Gly Thr Thr 35 40
45Ser Ser Ser Ile Pro Leu Tyr Arg Ser Ser Glu Glu Glu Lys Arg Val 50
55 60Thr Val Ile Lys Ala Pro His Tyr Pro
Gly Ile Gly Pro Val Asp Glu65 70 75
80Ser Gly Ile Pro Thr Ala Ile Arg Thr Thr Val Asp Arg Pro
Lys Asp 85 90 95Trp Tyr
Lys Thr Met Phe Lys Gln Ile His Met Val His Lys Pro Asp 100
105 110Asp Asp Thr Asp Met Tyr Asn Thr Pro
Tyr Thr Tyr Asn Ala Gly Leu 115 120
125Tyr Asn Pro Pro Tyr Ser Ala Gln Ser His Pro Ala Ala Lys Thr Gln
130 135 140Thr Tyr Arg Pro Leu Ser Lys
Ser His Ser Asp Asn Ser Pro Asn Ala145 150
155 160Phe Lys Asp Ala Ser Ser Pro Val Pro Pro Pro His
Val Pro Pro Pro 165 170
175Val Pro Pro Leu Arg Pro Arg Asp Arg Ser Ser Thr Glu Lys His Asp
180 185 190Trp Asp Pro Pro Asp Arg
Lys Val Asp Thr Arg Lys Phe Arg Ser Glu 195 200
205Pro Arg Ser Ile Phe Glu Tyr Glu Pro Gly Lys Ser Ser Ile
Leu Gln 210 215 220His Glu Arg Pro Thr
Asp Arg Ile Asn Pro Asp Asp Ile Asp Leu Glu225 230
235 240Asn Glu Pro Trp Tyr Lys Phe Phe Ser Glu
Leu Glu Phe Gly Arg Pro 245 250
255Pro Pro Lys Lys Pro Leu Asp Tyr Val Gln Asp His Ser Ser Gly Val
260 265 270Phe Asn Glu Ala Ser
Leu Tyr Gln Ser Ser Ile Asp Arg Ser Leu Glu 275
280 285Arg Pro Met Ser Ser Ala Ser Met Ala Ser Asp Phe
Arg Lys Arg Arg 290 295 300Lys Ser Glu
Pro Ala Val Gly Pro Pro Arg Gly Leu Gly Asp Gln Ser305
310 315 320Ala Ser Arg Thr Ser Pro Gly
Arg Val Asp Leu Pro Gly Ser Ser Thr 325
330 335Thr Leu Thr Lys Ser Phe Thr Ser Ser Ser Pro Ser
Ser Pro Ser Arg 340 345 350Ala
Lys Asp Arg Glu Ser Pro Arg Ser Tyr Ser Ser Thr Leu Thr Asp 355
360 365Met Gly Arg Ser Ala Pro Arg Glu Arg
Arg Gly Thr Pro Glu Lys Glu 370 375
380Lys Leu Pro Ala Lys Ala Val Tyr Asp Phe Lys Ala Gln Thr Ser Lys385
390 395 400Glu Leu Ser Phe
Lys Lys Gly Asp Thr Val Tyr Ile Leu Arg Lys Ile 405
410 415Asp Gln Asn Trp Tyr Glu Gly Glu His His
Gly Arg Val Gly Ile Phe 420 425
430Pro Ile Ser Tyr Val Glu Lys Leu Thr Pro Pro Glu Lys Ala Gln Pro
435 440 445Ala Arg Pro Pro Pro Pro Ala
Gln Pro Gly Glu Ile Gly Glu Ala Ile 450 455
460Ala Lys Tyr Asn Phe Asn Ala Asp Thr Asn Val Glu Leu Ser Leu
Arg465 470 475 480Lys Gly
Asp Arg Val Ile Leu Leu Lys Arg Val Asp Gln Asn Trp Tyr
485 490 495Glu Gly Lys Ile Pro Gly Thr
Asn Arg Gln Gly Ile Phe Pro Val Ser 500 505
510Tyr Val Glu Val Val Lys Lys Asn Thr Lys Gly Ala Glu Asp
Tyr Pro 515 520 525Asp Pro Pro Ile
Pro His Ser Tyr Ser Ser Asp Arg Ile His Ser Leu 530
535 540Ser Ser Asn Lys Pro Gln Arg Pro Val Phe Thr His
Glu Asn Ile Gln545 550 555
560Gly Gly Gly Glu Pro Phe Gln Ala Leu Tyr Asn Tyr Thr Pro Arg Asn
565 570 575Glu Asp Glu Leu Glu
Leu Arg Glu Ser Asp Val Ile Asp Val Met Glu 580
585 590Lys Cys Asp Asp Gly Trp Phe Val Gly Thr Ser Arg
Arg Thr Lys Phe 595 600 605Phe Gly
Thr Phe Pro Gly Asn Tyr Val Lys Arg Leu 610 615
62039179PRTHomo sapiens 39Met Ala Lys Thr Ala Met Ala Tyr Lys
Glu Lys Met Lys Glu Leu Ser1 5 10
15Met Leu Ser Leu Ile Cys Ser Cys Phe Tyr Pro Glu Pro Arg Asn
Ile 20 25 30Asn Ile Tyr Thr
Tyr Asp Asp Met Glu Val Lys Gln Ile Asn Lys Arg 35
40 45Ala Ser Gly Gln Ala Phe Glu Leu Ile Leu Lys Pro
Pro Ser Pro Ile 50 55 60Ser Glu Ala
Pro Arg Thr Leu Ala Ser Pro Lys Lys Lys Asp Leu Ser65 70
75 80Leu Glu Glu Ile Gln Lys Lys Leu
Glu Ala Ala Glu Glu Arg Arg Lys 85 90
95Ser Gln Glu Ala Gln Val Leu Lys Gln Leu Ala Glu Lys Arg
Glu His 100 105 110Glu Arg Glu
Val Leu Gln Lys Ala Leu Glu Glu Asn Asn Asn Phe Ser 115
120 125Lys Met Ala Glu Glu Lys Leu Ile Leu Lys Met
Glu Gln Ile Lys Glu 130 135 140Asn Arg
Glu Ala Asn Leu Ala Ala Ile Ile Glu Arg Leu Gln Glu Lys145
150 155 160Glu Arg His Ala Ala Glu Val
Arg Arg Asn Lys Glu Leu Gln Val Glu 165
170 175Leu Ser Gly40715PRTHomo sapiens 40Met Ser Lys Ala
His Lys Pro Trp Pro Tyr Arg Arg Arg Ser Gln Phe1 5
10 15Ser Ser Arg Lys Tyr Leu Lys Lys Glu Met
Asn Ser Phe Gln Gln Gln 20 25
30Pro Pro Pro Phe Gly Thr Val Pro Pro Gln Met Met Phe Pro Pro Asn
35 40 45Trp Gln Gly Ala Glu Lys Asp Ala
Ala Phe Leu Ala Lys Asp Phe Asn 50 55
60Phe Leu Thr Leu Asn Asn Gln Pro Pro Pro Gly Asn Arg Ser Gln Pro65
70 75 80Arg Ala Met Gly Pro
Glu Asn Asn Leu Tyr Ser Gln Tyr Glu Gln Lys 85
90 95Val Arg Pro Cys Ile Asp Leu Ile Asp Ser Leu
Arg Ala Leu Gly Val 100 105
110Glu Gln Asp Leu Ala Leu Pro Ala Ile Ala Val Ile Gly Asp Gln Ser
115 120 125Ser Gly Lys Ser Ser Val Leu
Glu Ala Leu Ser Gly Val Ala Leu Pro 130 135
140Arg Gly Ser Gly Ile Val Thr Arg Cys Pro Leu Val Leu Lys Leu
Lys145 150 155 160Lys Gln
Pro Cys Glu Ala Trp Ala Gly Arg Ile Ser Tyr Arg Asn Thr
165 170 175Glu Leu Glu Leu Gln Asp Pro
Gly Gln Val Glu Lys Glu Ile His Lys 180 185
190Ala Gln Asn Val Met Ala Gly Asn Gly Arg Gly Ile Ser His
Glu Leu 195 200 205Ile Ser Leu Glu
Ile Thr Ser Pro Glu Val Pro Asp Leu Thr Ile Ile 210
215 220Asp Leu Pro Gly Ile Thr Arg Val Ala Val Asp Asn
Gln Pro Arg Asp225 230 235
240Ile Gly Leu Gln Ile Lys Ala Leu Ile Lys Lys Tyr Ile Gln Arg Gln
245 250 255Gln Thr Ile Asn Leu
Val Val Val Pro Cys Asn Val Asp Ile Ala Thr 260
265 270Thr Glu Ala Leu Ser Met Ala His Glu Val Asp Pro
Glu Gly Asp Arg 275 280 285Thr Ile
Gly Ile Leu Thr Lys Pro Asp Leu Met Asp Arg Gly Thr Glu 290
295 300Lys Ser Val Met Asn Val Val Arg Asn Leu Thr
Tyr Pro Leu Lys Lys305 310 315
320Gly Tyr Met Ile Val Lys Cys Arg Gly Gln Gln Glu Ile Thr Asn Arg
325 330 335Leu Ser Leu Ala
Glu Ala Thr Lys Lys Glu Ile Thr Phe Phe Gln Thr 340
345 350His Pro Tyr Phe Arg Val Leu Leu Glu Glu Gly
Ser Ala Thr Val Pro 355 360 365Arg
Leu Ala Glu Arg Leu Thr Thr Glu Leu Ile Met His Ile Gln Lys 370
375 380Ser Leu Pro Leu Leu Glu Gly Gln Ile Arg
Glu Ser His Gln Lys Ala385 390 395
400Thr Glu Glu Leu Arg Arg Cys Gly Ala Asp Ile Pro Ser Gln Glu
Ala 405 410 415Asp Lys Met
Phe Phe Leu Ile Glu Lys Ile Lys Met Phe Asn Gln Asp 420
425 430Ile Glu Lys Leu Val Glu Gly Glu Glu Val
Val Arg Glu Asn Glu Thr 435 440
445Arg Leu Tyr Asn Lys Ile Arg Glu Asp Phe Lys Asn Trp Val Gly Ile 450
455 460Leu Ala Thr Asn Thr Gln Lys Val
Lys Asn Ile Ile His Glu Glu Val465 470
475 480Glu Lys Tyr Glu Lys Gln Tyr Arg Gly Lys Glu Leu
Leu Gly Phe Val 485 490
495Asn Tyr Lys Thr Phe Glu Ile Ile Val His Gln Tyr Ile Gln Gln Leu
500 505 510Val Glu Pro Ala Leu Ser
Met Leu Gln Lys Ala Met Glu Ile Ile Gln 515 520
525Gln Ala Phe Ile Asn Val Ala Lys Lys His Phe Gly Glu Phe
Phe Asn 530 535 540Leu Asn Gln Thr Val
Gln Ser Thr Ile Glu Asp Ile Lys Val Lys His545 550
555 560Thr Ala Lys Ala Glu Asn Met Ile Gln Leu
Gln Phe Arg Met Glu Gln 565 570
575Met Val Phe Cys Gln Asp Gln Ile Tyr Ser Val Val Leu Lys Lys Val
580 585 590Arg Glu Glu Ile Phe
Asn Pro Leu Gly Thr Pro Ser Gln Asn Met Lys 595
600 605Leu Asn Ser His Phe Pro Ser Asn Glu Ser Ser Val
Ser Ser Phe Thr 610 615 620Glu Ile Gly
Ile His Leu Asn Ala Tyr Phe Leu Glu Thr Ser Lys Arg625
630 635 640Leu Ala Asn Gln Ile Pro Phe
Ile Ile Gln Tyr Phe Met Leu Arg Glu 645
650 655Asn Gly Asp Ser Leu Gln Lys Ala Met Met Gln Ile
Leu Gln Glu Lys 660 665 670Asn
Arg Tyr Ser Trp Leu Leu Gln Glu Gln Ser Glu Thr Ala Thr Lys 675
680 685Arg Arg Ile Leu Lys Glu Arg Ile Tyr
Arg Leu Thr Gln Ala Arg His 690 695
700Ala Leu Cys Gln Phe Ser Ser Lys Glu Ile His705 710
71541503PRTHomo sapiens 41Met Ala Leu Ala Pro Glu Arg Ala Ala
Pro Arg Val Leu Phe Gly Glu1 5 10
15Trp Leu Leu Gly Glu Ile Ser Ser Gly Cys Tyr Glu Gly Leu Gln
Trp 20 25 30Leu Asp Glu Ala
Arg Thr Cys Phe Arg Val Pro Trp Lys His Phe Ala 35
40 45Arg Lys Asp Leu Ser Glu Ala Asp Ala Arg Ile Phe
Lys Ala Trp Ala 50 55 60Val Ala Arg
Gly Arg Trp Pro Pro Ser Ser Arg Gly Gly Gly Pro Pro65 70
75 80Pro Glu Ala Glu Thr Ala Glu Arg
Ala Gly Trp Lys Thr Asn Phe Arg 85 90
95Cys Ala Leu Arg Ser Thr Arg Arg Phe Val Met Leu Arg Asp
Asn Ser 100 105 110Gly Asp Pro
Ala Asp Pro His Lys Val Tyr Ala Leu Ser Arg Glu Leu 115
120 125Cys Trp Arg Glu Gly Pro Gly Thr Asp Gln Thr
Glu Ala Glu Ala Pro 130 135 140Ala Ala
Val Pro Pro Pro Gln Gly Gly Pro Pro Gly Pro Phe Leu Ala145
150 155 160His Thr His Ala Gly Leu Gln
Ala Pro Gly Pro Leu Pro Ala Pro Ala 165
170 175Gly Asp Lys Gly Asp Leu Leu Leu Gln Ala Val Gln
Gln Ser Cys Leu 180 185 190Ala
Asp His Leu Leu Thr Ala Ser Trp Gly Ala Asp Pro Val Pro Thr 195
200 205Lys Ala Pro Gly Glu Gly Gln Glu Gly
Leu Pro Leu Thr Gly Ala Cys 210 215
220Ala Gly Gly Pro Gly Leu Pro Ala Gly Glu Leu Tyr Gly Trp Ala Val225
230 235 240Glu Thr Thr Pro
Ser Pro Gly Pro Gln Pro Ala Ala Leu Thr Thr Gly 245
250 255Glu Ala Ala Ala Pro Glu Ser Pro His Gln
Ala Glu Pro Tyr Leu Ser 260 265
270Pro Ser Pro Ser Ala Cys Thr Ala Val Gln Glu Pro Ser Pro Gly Ala
275 280 285Leu Asp Val Thr Ile Met Tyr
Lys Gly Arg Thr Val Leu Gln Lys Val 290 295
300Val Gly His Pro Ser Cys Thr Phe Leu Tyr Gly Pro Pro Asp Pro
Ala305 310 315 320Val Arg
Ala Thr Asp Pro Gln Gln Val Ala Phe Pro Ser Pro Ala Glu
325 330 335Leu Pro Asp Gln Lys Gln Leu
Arg Tyr Thr Glu Glu Leu Leu Arg His 340 345
350Val Ala Pro Gly Leu His Leu Glu Leu Arg Gly Pro Gln Leu
Trp Ala 355 360 365Arg Arg Met Gly
Lys Cys Lys Val Tyr Trp Glu Val Gly Gly Pro Pro 370
375 380Gly Ser Ala Ser Pro Ser Thr Pro Ala Cys Leu Leu
Pro Arg Asn Cys385 390 395
400Asp Thr Pro Ile Phe Asp Phe Arg Val Phe Phe Gln Glu Leu Val Glu
405 410 415Phe Arg Ala Arg Gln
Arg Arg Gly Ser Pro Arg Tyr Thr Ile Tyr Leu 420
425 430Gly Phe Gly Gln Asp Leu Ser Ala Gly Arg Pro Lys
Glu Lys Ser Leu 435 440 445Val Leu
Val Lys Leu Glu Pro Trp Leu Cys Arg Val His Leu Glu Gly 450
455 460Thr Gln Arg Glu Gly Val Ser Ser Leu Asp Ser
Ser Ser Leu Ser Leu465 470 475
480Cys Leu Ser Ser Ala Asn Ser Leu Tyr Asp Asp Ile Glu Cys Phe Leu
485 490 495Met Glu Leu Glu
Gln Pro Ala 50042591PRTHomo sapiens 42Met Ala Pro Glu Ile Asn
Leu Pro Gly Pro Met Ser Leu Ile Asp Asn1 5
10 15Thr Lys Gly Gln Leu Val Val Asn Pro Glu Ala Leu
Lys Ile Leu Ser 20 25 30Ala
Ile Thr Gln Pro Val Val Val Val Ala Ile Val Gly Leu Tyr Arg 35
40 45Thr Gly Lys Ser Tyr Leu Met Asn Lys
Leu Ala Gly Lys Lys Asn Gly 50 55
60Phe Ser Leu Gly Ser Thr Val Lys Ser His Thr Lys Gly Ile Trp Met65
70 75 80Trp Cys Val Pro His
Pro Lys Lys Pro Glu His Thr Leu Val Leu Leu 85
90 95Asp Thr Glu Gly Leu Gly Asp Ile Glu Lys Gly
Asp Asn Glu Asn Asp 100 105
110Ser Trp Ile Phe Ala Leu Ala Ile Leu Leu Ser Ser Thr Phe Val Tyr
115 120 125Asn Ser Met Gly Thr Ile Asn
Gln Gln Ala Met Asp Gln Leu His Tyr 130 135
140Val Thr Glu Leu Thr Asp Arg Ile Lys Ala Asn Ser Ser Pro Gly
Asn145 150 155 160Asn Ser
Val Asp Asp Ser Ala Asp Phe Val Ser Phe Phe Pro Ala Phe
165 170 175Val Trp Thr Leu Arg Asp Phe
Thr Leu Glu Leu Glu Val Asp Gly Glu 180 185
190Pro Ile Thr Ala Asp Asp Tyr Leu Glu Leu Ser Leu Lys Leu
Arg Lys 195 200 205Gly Thr Asp Lys
Lys Ser Lys Ser Phe Asn Asp Pro Arg Leu Cys Ile 210
215 220Arg Lys Phe Phe Pro Lys Arg Lys Cys Phe Val Phe
Asp Trp Pro Ala225 230 235
240Pro Lys Lys Tyr Leu Ala His Leu Glu Gln Leu Lys Glu Glu Glu Leu
245 250 255Asn Pro Asp Phe Ile
Glu Gln Val Ala Glu Phe Cys Ser Tyr Ile Leu 260
265 270Ser His Ser Asn Val Lys Thr Leu Ser Gly Gly Ile
Pro Val Asn Gly 275 280 285Pro Arg
Leu Glu Ser Leu Val Leu Thr Tyr Val Asn Ala Ile Ser Ser 290
295 300Gly Asp Leu Pro Cys Met Glu Asn Ala Val Leu
Ala Leu Ala Gln Ile305 310 315
320Glu Asn Ser Ala Ala Val Glu Lys Ala Ile Ala His Tyr Glu Gln Gln
325 330 335Met Gly Gln Lys
Val Gln Leu Pro Thr Glu Thr Leu Gln Glu Leu Leu 340
345 350Asp Leu His Arg Asp Ser Glu Arg Glu Ala Ile
Glu Val Phe Met Lys 355 360 365Asn
Ser Phe Lys Asp Val Asp Gln Met Phe Gln Arg Lys Leu Gly Ala 370
375 380Gln Leu Glu Ala Arg Arg Asp Asp Phe Cys
Lys Gln Asn Ser Lys Ala385 390 395
400Ser Ser Asp Cys Cys Met Ala Leu Leu Gln Asp Ile Phe Gly Pro
Leu 405 410 415Glu Glu Asp
Val Lys Gln Gly Thr Phe Ser Lys Pro Gly Gly Tyr Arg 420
425 430Leu Phe Thr Gln Lys Leu Gln Glu Leu Lys
Asn Lys Tyr Tyr Gln Val 435 440
445Pro Arg Lys Gly Ile Gln Ala Lys Glu Val Leu Lys Lys Tyr Leu Glu 450
455 460Ser Lys Glu Asp Val Ala Asp Ala
Leu Leu Gln Thr Asp Gln Ser Leu465 470
475 480Ser Glu Lys Glu Lys Ala Ile Glu Val Glu Arg Ile
Lys Ala Glu Ser 485 490
495Ala Glu Ala Ala Lys Lys Met Leu Glu Glu Ile Gln Lys Lys Asn Glu
500 505 510Glu Met Met Glu Gln Lys
Glu Lys Ser Tyr Gln Glu His Val Lys Gln 515 520
525Leu Thr Glu Lys Met Glu Arg Asp Arg Ala Gln Leu Met Ala
Glu Gln 530 535 540Glu Lys Thr Leu Ala
Leu Lys Leu Gln Glu Gln Glu Arg Leu Leu Lys545 550
555 560Glu Gly Phe Glu Asn Glu Ser Lys Arg Leu
Gln Lys Asp Ile Trp Asp 565 570
575Ile Gln Met Arg Ser Lys Ser Leu Glu Pro Ile Cys Asn Ile Leu
580 585 59043658PRTHomo sapiens
43Met Glu Lys Ala Ser Gly Arg Gln Ser Ile Ala Leu Ser Thr Val Glu1
5 10 15Thr Gly Thr Val Asn Pro
Gly Leu Glu Leu Met Glu Lys Glu Val Glu 20 25
30Pro Glu Gly Ser Lys Arg Thr Asp Ala Gln Gly His Ser
Leu Gly Asp 35 40 45Gly Leu Gly
Pro Ser Thr Tyr Gln Arg Arg Ser Arg Trp Pro Phe Ser 50
55 60Lys Ala Arg Ser Phe Cys Lys Thr His Ala Arg Leu
Phe Lys Lys Ile65 70 75
80Leu Leu Gly Leu Leu Cys Leu Ala Tyr Ala Ala Tyr Leu Leu Ala Ala
85 90 95Cys Ile Leu Asn Phe Gln
Arg Ala Leu Ala Leu Phe Val Ile Thr Cys 100
105 110Leu Val Ile Phe Val Leu Val His Ser Phe Leu Lys
Lys Leu Leu Gly 115 120 125Lys Lys
Leu Thr Arg Cys Leu Lys Pro Phe Glu Asn Ser Arg Leu Arg 130
135 140Leu Trp Thr Lys Trp Val Phe Ala Gly Val Ser
Leu Val Gly Leu Ile145 150 155
160Leu Trp Leu Ala Leu Asp Thr Ala Gln Arg Pro Glu Gln Leu Ile Pro
165 170 175Phe Ala Gly Ile
Cys Met Phe Ile Leu Ile Leu Phe Ala Cys Ser Lys 180
185 190His His Ser Ala Val Ser Trp Arg Thr Val Phe
Ser Gly Leu Gly Leu 195 200 205Gln
Phe Val Phe Gly Ile Leu Val Ile Arg Thr Asp Leu Gly Tyr Thr 210
215 220Val Phe Gln Trp Leu Gly Glu Gln Val Gln
Ile Phe Leu Asn Tyr Thr225 230 235
240Val Ala Gly Ser Ser Phe Val Phe Gly Asp Thr Leu Val Lys Asp
Val 245 250 255Phe Ala Phe
Gln Ala Leu Pro Ile Ile Ile Phe Phe Gly Cys Val Val 260
265 270Ser Ile Leu Tyr Tyr Leu Gly Leu Val Gln
Trp Val Val Gln Lys Val 275 280
285Ala Trp Phe Leu Gln Ile Thr Met Gly Thr Thr Ala Thr Glu Thr Leu 290
295 300Ala Val Ala Gly Asn Ile Phe Val
Gly Met Thr Glu Ala Pro Leu Leu305 310
315 320Ile Arg Pro Tyr Leu Gly Asp Met Thr Leu Ser Glu
Ile His Ala Val 325 330
335Met Thr Gly Gly Phe Ala Thr Ile Ser Gly Thr Val Leu Gly Ala Phe
340 345 350Ile Ala Phe Gly Val Asp
Ala Ser Ser Leu Ile Ser Ala Ser Val Met 355 360
365Ala Ala Pro Cys Ala Leu Ala Ser Ser Lys Leu Ala Tyr Pro
Glu Val 370 375 380Glu Glu Ser Lys Phe
Lys Ser Glu Glu Gly Val Lys Leu Pro Arg Gly385 390
395 400Lys Glu Arg Asn Val Leu Glu Ala Ala Ser
Asn Gly Ala Val Asp Ala 405 410
415Ile Gly Leu Ala Thr Asn Val Ala Ala Asn Leu Ile Ala Phe Leu Ala
420 425 430Val Leu Ala Phe Ile
Asn Ala Ala Leu Ser Trp Leu Gly Glu Leu Val 435
440 445Asp Ile Gln Gly Leu Thr Phe Gln Val Ile Cys Ser
Tyr Leu Leu Arg 450 455 460Pro Met Val
Phe Met Met Gly Val Glu Trp Thr Asp Cys Pro Met Val465
470 475 480Ala Glu Met Val Gly Ile Lys
Phe Phe Ile Asn Glu Phe Val Ala Tyr 485
490 495Gln Gln Leu Ser Gln Tyr Lys Asn Lys Arg Leu Ser
Gly Met Glu Glu 500 505 510Trp
Ile Glu Gly Glu Lys Gln Trp Ile Ser Val Arg Ala Glu Ile Ile 515
520 525Thr Thr Phe Ser Leu Cys Gly Phe Ala
Asn Leu Ser Ser Ile Gly Ile 530 535
540Thr Leu Gly Gly Leu Thr Ser Ile Val Pro His Arg Lys Ser Asp Leu545
550 555 560Ser Lys Val Val
Val Arg Ala Leu Phe Thr Gly Ala Cys Val Ser Leu 565
570 575Ile Ser Ala Cys Met Ala Gly Ile Leu Tyr
Val Pro Arg Gly Ala Glu 580 585
590Ala Asp Cys Val Ser Phe Pro Asn Thr Ser Phe Thr Asn Arg Thr Tyr
595 600 605Glu Thr Tyr Met Cys Cys Arg
Gly Leu Phe Gln Ser Thr Ser Leu Asn 610 615
620Gly Thr Asn Pro Pro Ser Phe Ser Gly Pro Trp Glu Asp Lys Glu
Phe625 630 635 640Ser Ala
Met Ala Leu Thr Asn Cys Cys Gly Phe Tyr Asn Asn Thr Val
645 650 655Cys Ala44247PRTHomo sapiens
44Met Thr Ile Leu Phe Leu Thr Met Val Ile Ser Tyr Phe Gly Cys Met1
5 10 15Lys Ala Ala Pro Met Lys
Glu Ala Asn Ile Arg Gly Gln Gly Gly Leu 20 25
30Ala Tyr Pro Gly Val Arg Thr His Gly Thr Leu Glu Ser
Val Asn Gly 35 40 45Pro Lys Ala
Gly Ser Arg Gly Leu Thr Ser Leu Ala Asp Thr Phe Glu 50
55 60His Val Ile Glu Glu Leu Leu Asp Glu Asp Gln Lys
Val Arg Pro Asn65 70 75
80Glu Glu Asn Asn Lys Asp Ala Asp Leu Tyr Thr Ser Arg Val Met Leu
85 90 95Ser Ser Gln Val Pro Leu
Glu Pro Pro Leu Leu Phe Leu Leu Glu Glu 100
105 110Tyr Lys Asn Tyr Leu Asp Ala Ala Asn Met Ser Met
Arg Val Arg Arg 115 120 125His Ser
Asp Pro Ala Arg Arg Gly Glu Leu Ser Val Cys Asp Ser Ile 130
135 140Ser Glu Trp Val Thr Ala Ala Asp Lys Lys Thr
Ala Val Asp Met Ser145 150 155
160Gly Gly Thr Val Thr Val Leu Glu Lys Val Pro Val Ser Lys Gly Gln
165 170 175Leu Lys Gln Tyr
Phe Tyr Glu Thr Lys Cys Asn Pro Met Gly Tyr Thr 180
185 190Lys Glu Gly Cys Arg Gly Ile Asp Lys Arg His
Trp Asn Ser Gln Cys 195 200 205Arg
Thr Thr Gln Ser Tyr Val Arg Ala Leu Thr Met Asp Ser Lys Lys 210
215 220Arg Ile Gly Trp Arg Phe Ile Arg Ile Asp
Thr Ser Cys Val Cys Thr225 230 235
240Leu Thr Ile Lys Arg Gly Arg 245451112PRTHomo
sapiens 45Met Arg Arg Asp Glu Arg Asp Ala Lys Ala Met Arg Ser Leu Gln
Pro1 5 10 15Pro Asp Gly
Ala Gly Ser Pro Pro Glu Ser Leu Arg Asn Gly Tyr Val 20
25 30Lys Ser Cys Val Ser Pro Leu Arg Gln Asp
Pro Pro Arg Gly Phe Phe 35 40
45Phe His Leu Cys Arg Phe Cys Asn Val Glu Leu Arg Pro Pro Pro Ala 50
55 60Ser Pro Gln Gln Pro Arg Arg Cys Ser
Pro Phe Cys Arg Ala Arg Leu65 70 75
80Ser Leu Gly Ala Leu Ala Ala Phe Val Leu Ala Leu Leu Leu
Gly Ala 85 90 95Glu Pro
Glu Ser Trp Ala Ala Gly Ala Ala Trp Leu Arg Thr Leu Leu 100
105 110Ser Val Cys Ser His Ser Leu Ser Pro
Leu Phe Ser Ile Ala Cys Ala 115 120
125Phe Phe Phe Leu Thr Cys Phe Leu Thr Arg Thr Lys Arg Gly Pro Gly
130 135 140Pro Gly Arg Ser Cys Gly Ser
Trp Trp Leu Leu Ala Leu Pro Ala Cys145 150
155 160Cys Tyr Leu Gly Asp Phe Leu Val Trp Gln Trp Trp
Ser Trp Pro Trp 165 170
175Gly Asp Gly Asp Ala Gly Ser Ala Ala Pro His Thr Pro Pro Glu Ala
180 185 190Ala Ala Gly Arg Leu Leu
Leu Val Leu Ser Cys Val Gly Leu Leu Leu 195 200
205Thr Leu Ala His Pro Leu Arg Leu Arg His Cys Val Leu Val
Leu Leu 210 215 220Leu Ala Ser Phe Val
Trp Trp Val Ser Phe Thr Ser Leu Gly Ser Leu225 230
235 240Pro Ser Ala Leu Arg Pro Leu Leu Ser Gly
Leu Val Gly Gly Ala Gly 245 250
255Cys Leu Leu Ala Leu Gly Leu Asp His Phe Phe Gln Ile Arg Glu Ala
260 265 270Pro Leu His Pro Arg
Leu Ser Ser Ala Ala Glu Glu Lys Val Pro Val 275
280 285Ile Arg Pro Arg Arg Arg Ser Ser Cys Val Ser Leu
Gly Glu Thr Ala 290 295 300Ala Ser Tyr
Tyr Gly Ser Cys Lys Ile Phe Arg Arg Pro Ser Leu Pro305
310 315 320Cys Ile Ser Arg Glu Gln Met
Ile Leu Trp Asp Trp Asp Leu Lys Gln 325
330 335Trp Tyr Lys Pro His Tyr Gln Asn Ser Gly Gly Gly
Asn Gly Val Asp 340 345 350Leu
Ser Val Leu Asn Glu Ala Arg Asn Met Val Ser Asp Leu Leu Thr 355
360 365Asp Pro Ser Leu Pro Pro Gln Val Ile
Ser Ser Leu Arg Ser Ile Ser 370 375
380Ser Leu Met Gly Ala Phe Ser Gly Ser Cys Arg Pro Lys Ile Asn Pro385
390 395 400Leu Thr Pro Phe
Pro Gly Phe Tyr Pro Cys Ser Glu Ile Glu Asp Pro 405
410 415Ala Glu Lys Gly Asp Arg Lys Leu Asn Lys
Gly Leu Asn Arg Asn Ser 420 425
430Leu Pro Thr Pro Gln Leu Arg Arg Ser Ser Gly Thr Ser Gly Leu Leu
435 440 445Pro Val Glu Gln Ser Ser Arg
Trp Asp Arg Asn Asn Gly Lys Arg Pro 450 455
460His Gln Glu Phe Gly Ile Ser Ser Gln Gly Cys Tyr Leu Asn Gly
Pro465 470 475 480Phe Asn
Ser Asn Leu Leu Thr Ile Pro Lys Gln Arg Ser Ser Ser Val
485 490 495Ser Leu Thr His His Val Gly
Leu Arg Arg Ala Gly Val Leu Ser Ser 500 505
510Leu Ser Pro Val Asn Ser Ser Asn His Gly Pro Val Ser Thr
Gly Ser 515 520 525Leu Thr Asn Arg
Ser Pro Ile Glu Phe Pro Asp Thr Ala Asp Phe Leu 530
535 540Asn Lys Pro Ser Val Ile Leu Gln Arg Ser Leu Gly
Asn Ala Pro Asn545 550 555
560Thr Pro Asp Phe Tyr Gln Gln Leu Arg Asn Ser Asp Ser Asn Leu Cys
565 570 575Asn Ser Cys Gly His
Gln Met Leu Lys Tyr Val Ser Thr Ser Glu Ser 580
585 590Asp Gly Thr Asp Cys Cys Ser Gly Lys Ser Gly Glu
Glu Glu Asn Ile 595 600 605Phe Ser
Lys Glu Ser Phe Lys Leu Met Glu Thr Gln Gln Glu Glu Glu 610
615 620Thr Glu Lys Lys Asp Ser Arg Lys Leu Phe Gln
Glu Gly Asp Lys Trp625 630 635
640Leu Thr Glu Glu Ala Gln Ser Glu Gln Gln Thr Asn Ile Glu Gln Glu
645 650 655Val Ser Leu Asp
Leu Ile Leu Val Glu Glu Tyr Asp Ser Leu Ile Glu 660
665 670Lys Met Ser Asn Trp Asn Phe Pro Ile Phe Glu
Leu Val Glu Lys Met 675 680 685Gly
Glu Lys Ser Gly Arg Ile Leu Ser Gln Val Met Tyr Thr Leu Phe 690
695 700Gln Asp Thr Gly Leu Leu Glu Ile Phe Lys
Ile Pro Thr Gln Gln Phe705 710 715
720Met Asn Tyr Phe Arg Ala Leu Glu Asn Gly Tyr Arg Asp Ile Pro
Tyr 725 730 735His Asn Arg
Ile His Ala Thr Asp Val Leu His Ala Val Trp Tyr Leu 740
745 750Thr Thr Arg Pro Val Pro Gly Leu Gln Gln
Ile His Asn Gly Cys Gly 755 760
765Thr Gly Asn Glu Thr Asp Ser Asp Gly Arg Ile Asn His Gly Arg Ile 770
775 780Ala Tyr Ile Ser Ser Lys Ser Cys
Ser Asn Pro Asp Glu Ser Tyr Gly785 790
795 800Cys Leu Ser Ser Asn Ile Pro Ala Leu Glu Leu Met
Ala Leu Tyr Val 805 810
815Ala Ala Ala Met His Asp Tyr Asp His Pro Gly Arg Thr Asn Ala Phe
820 825 830Leu Val Ala Thr Asn Ala
Pro Gln Ala Val Leu Tyr Asn Asp Arg Ser 835 840
845Val Leu Glu Asn His His Ala Ala Ser Ala Trp Asn Leu Tyr
Leu Ser 850 855 860Arg Pro Glu Tyr Asn
Phe Leu Leu His Leu Asp His Val Glu Phe Lys865 870
875 880Arg Phe Arg Phe Leu Val Ile Glu Ala Ile
Leu Ala Thr Asp Leu Lys 885 890
895Lys His Phe Asp Phe Leu Ala Glu Phe Asn Ala Lys Ala Asn Asp Val
900 905 910Asn Ser Asn Gly Ile
Glu Trp Ser Asn Glu Asn Asp Arg Leu Leu Val 915
920 925Cys Gln Val Cys Ile Lys Leu Ala Asp Ile Asn Gly
Pro Ala Lys Val 930 935 940Arg Asp Leu
His Leu Lys Trp Thr Glu Gly Ile Val Asn Glu Phe Tyr945
950 955 960Glu Gln Gly Asp Glu Glu Ala
Asn Leu Gly Leu Pro Ile Ser Pro Phe 965
970 975Met Asp Arg Ser Ser Pro Gln Leu Ala Lys Leu Gln
Glu Ser Phe Ile 980 985 990Thr
His Ile Val Gly Pro Leu Cys Asn Ser Tyr Asp Ala Ala Gly Leu 995
1000 1005Leu Pro Gly Gln Trp Leu Glu Ala
Glu Glu Asp Asn Asp Thr Glu 1010 1015
1020Ser Gly Asp Asp Glu Asp Gly Glu Glu Leu Asp Thr Glu Asp Glu
1025 1030 1035Glu Met Glu Asn Asn Leu
Asn Pro Lys Pro Pro Arg Arg Lys Ser 1040 1045
1050Arg Arg Arg Ile Phe Cys Gln Leu Met His His Leu Thr Glu
Asn 1055 1060 1065His Lys Ile Trp Lys
Glu Ile Val Glu Glu Glu Glu Lys Cys Lys 1070 1075
1080Ala Asp Gly Asn Lys Leu Gln Val Glu Asn Ser Ser Leu
Pro Gln 1085 1090 1095Ala Asp Glu Ile
Gln Val Ile Glu Glu Ala Asp Glu Glu Glu 1100 1105
111046538PRTHomo sapiens 46Met Phe Phe Leu Tyr Thr Asp Phe
Phe Leu Ser Leu Val Ala Val Pro1 5 10
15Ala Ala Ala Pro Val Cys Gln Pro Lys Ser Ala Thr Asn Gly
Gln Pro 20 25 30Pro Ala Pro
Ala Pro Thr Pro Thr Pro Arg Leu Ser Ile Ser Ser Arg 35
40 45Ala Thr Val Val Ala Arg Met Glu Gly Thr Ser
Gln Gly Gly Leu Gln 50 55 60Thr Val
Met Lys Trp Lys Thr Val Val Ala Ile Phe Val Val Val Val65
70 75 80Val Tyr Leu Val Thr Gly Gly
Leu Val Phe Arg Ala Leu Glu Gln Pro 85 90
95Phe Glu Ser Ser Gln Lys Asn Thr Ile Ala Leu Glu Lys
Ala Glu Phe 100 105 110Leu Arg
Asp His Val Cys Val Ser Pro Gln Glu Leu Glu Thr Leu Ile 115
120 125Gln His Ala Leu Asp Ala Asp Asn Ala Gly
Val Ser Pro Ile Gly Asn 130 135 140Ser
Ser Asn Asn Ser Ser His Trp Asp Leu Gly Ser Ala Phe Phe Phe145
150 155 160Ala Gly Thr Val Ile Thr
Thr Ile Gly Tyr Gly Asn Ile Ala Pro Ser 165
170 175Thr Glu Gly Gly Lys Ile Phe Cys Ile Leu Tyr Ala
Ile Phe Gly Ile 180 185 190Pro
Leu Phe Gly Phe Leu Leu Ala Gly Ile Gly Asp Gln Leu Gly Thr 195
200 205Ile Phe Gly Lys Ser Ile Ala Arg Val
Glu Lys Val Phe Arg Lys Lys 210 215
220Gln Val Ser Gln Thr Lys Ile Arg Val Ile Ser Thr Ile Leu Phe Ile225
230 235 240Leu Ala Gly Cys
Ile Val Phe Val Thr Ile Pro Ala Val Ile Phe Lys 245
250 255Tyr Ile Glu Gly Trp Thr Ala Leu Glu Ser
Ile Tyr Phe Val Val Val 260 265
270Thr Leu Thr Thr Val Gly Phe Gly Asp Phe Val Ala Gly Gly Asn Ala
275 280 285Gly Ile Asn Tyr Arg Glu Trp
Tyr Lys Pro Leu Val Trp Phe Trp Ile 290 295
300Leu Val Gly Leu Ala Tyr Phe Ala Ala Val Leu Ser Met Ile Gly
Asp305 310 315 320Trp Leu
Arg Val Leu Ser Lys Lys Thr Lys Glu Glu Val Gly Glu Ile
325 330 335Lys Ala His Ala Ala Glu Trp
Lys Ala Asn Val Thr Ala Glu Phe Arg 340 345
350Glu Thr Arg Arg Arg Leu Ser Val Glu Ile His Asp Lys Leu
Gln Arg 355 360 365Ala Ala Thr Ile
Arg Ser Met Glu Arg Arg Arg Leu Gly Leu Asp Gln 370
375 380Arg Ala His Ser Leu Asp Met Leu Ser Pro Glu Lys
Arg Ser Val Phe385 390 395
400Ala Ala Leu Asp Thr Gly Arg Phe Lys Ala Ser Ser Gln Glu Ser Ile
405 410 415Asn Asn Arg Pro Asn
Asn Leu Arg Leu Lys Gly Pro Glu Gln Leu Asn 420
425 430Lys His Gly Gln Gly Ala Ser Glu Asp Asn Ile Ile
Asn Lys Phe Gly 435 440 445Ser Thr
Ser Arg Leu Thr Lys Arg Lys Asn Lys Asp Leu Lys Lys Thr 450
455 460Leu Pro Glu Asp Val Gln Lys Ile Tyr Lys Thr
Phe Arg Asn Tyr Ser465 470 475
480Leu Asp Glu Glu Lys Lys Glu Glu Glu Thr Glu Lys Met Cys Asn Ser
485 490 495Asp Asn Ser Ser
Thr Ala Met Leu Thr Asp Cys Ile Gln Gln His Ala 500
505 510Glu Leu Glu Asn Gly Met Ile Pro Thr Asp Thr
Lys Asp Arg Glu Pro 515 520 525Glu
Asn Asn Ser Leu Leu Glu Asp Arg Asn 530
53547796PRTHomo sapiens 47Met Leu Arg Gly Gly Arg Arg Gly Gln Leu Gly Trp
His Ser Trp Ala1 5 10
15Ala Gly Pro Gly Ser Leu Leu Ala Trp Leu Ile Leu Ala Ser Ala Gly
20 25 30Ala Ala Pro Cys Pro Asp Ala
Cys Cys Pro His Gly Ser Ser Gly Leu 35 40
45Arg Cys Thr Arg Asp Gly Ala Leu Asp Ser Leu His His Leu Pro
Gly 50 55 60Ala Glu Asn Leu Thr Glu
Leu Tyr Ile Glu Asn Gln Gln His Leu Gln65 70
75 80His Leu Glu Leu Arg Asp Leu Arg Gly Leu Gly
Glu Leu Arg Asn Leu 85 90
95Thr Ile Val Lys Ser Gly Leu Arg Phe Val Ala Pro Asp Ala Phe His
100 105 110Phe Thr Pro Arg Leu Ser
Arg Leu Asn Leu Ser Phe Asn Ala Leu Glu 115 120
125Ser Leu Ser Trp Lys Thr Val Gln Gly Leu Ser Leu Gln Glu
Leu Val 130 135 140Leu Ser Gly Asn Pro
Leu His Cys Ser Cys Ala Leu Arg Trp Leu Gln145 150
155 160Arg Trp Glu Glu Glu Gly Leu Gly Gly Val
Pro Glu Gln Lys Leu Gln 165 170
175Cys His Gly Gln Gly Pro Leu Ala His Met Pro Asn Ala Ser Cys Gly
180 185 190Val Pro Thr Leu Lys
Val Gln Val Pro Asn Ala Ser Val Asp Val Gly 195
200 205Asp Asp Val Leu Leu Arg Cys Gln Val Glu Gly Arg
Gly Leu Glu Gln 210 215 220Ala Gly Trp
Ile Leu Thr Glu Leu Glu Gln Ser Ala Thr Val Met Lys225
230 235 240Ser Gly Gly Leu Pro Ser Leu
Gly Leu Thr Leu Ala Asn Val Thr Ser 245
250 255Asp Leu Asn Arg Lys Asn Val Thr Cys Trp Ala Glu
Asn Asp Val Gly 260 265 270Arg
Ala Glu Val Ser Val Gln Val Asn Val Ser Phe Pro Ala Ser Val 275
280 285Gln Leu His Thr Ala Val Glu Met His
His Trp Cys Ile Pro Phe Ser 290 295
300Val Asp Gly Gln Pro Ala Pro Ser Leu Arg Trp Leu Phe Asn Gly Ser305
310 315 320Val Leu Asn Glu
Thr Ser Phe Ile Phe Thr Glu Phe Leu Glu Pro Ala 325
330 335Ala Asn Glu Thr Val Arg His Gly Cys Leu
Arg Leu Asn Gln Pro Thr 340 345
350His Val Asn Asn Gly Asn Tyr Thr Leu Leu Ala Ala Asn Pro Phe Gly
355 360 365Gln Ala Ser Ala Ser Ile Met
Ala Ala Phe Met Asp Asn Pro Phe Glu 370 375
380Phe Asn Pro Glu Asp Pro Ile Pro Val Ser Phe Ser Pro Val Asp
Thr385 390 395 400Asn Ser
Thr Ser Gly Asp Pro Val Glu Lys Lys Asp Glu Thr Pro Phe
405 410 415Gly Val Ser Val Ala Val Gly
Leu Ala Val Phe Ala Cys Leu Phe Leu 420 425
430Ser Thr Leu Leu Leu Val Leu Asn Lys Cys Gly Arg Arg Asn
Lys Phe 435 440 445Gly Ile Asn Arg
Pro Ala Val Leu Ala Pro Glu Asp Gly Leu Ala Met 450
455 460Ser Leu His Phe Met Thr Leu Gly Gly Ser Ser Leu
Ser Pro Thr Glu465 470 475
480Gly Lys Gly Ser Gly Leu Gln Gly His Ile Ile Glu Asn Pro Gln Tyr
485 490 495Phe Ser Asp Ala Cys
Val His His Ile Lys Arg Arg Asp Ile Val Leu 500
505 510Lys Trp Glu Leu Gly Glu Gly Ala Phe Gly Lys Val
Phe Leu Ala Glu 515 520 525Cys His
Asn Leu Leu Pro Glu Gln Asp Lys Met Leu Val Ala Val Lys 530
535 540Ala Leu Lys Glu Ala Ser Glu Ser Ala Arg Gln
Asp Phe Gln Arg Glu545 550 555
560Ala Glu Leu Leu Thr Met Leu Gln His Gln His Ile Val Arg Phe Phe
565 570 575Gly Val Cys Thr
Glu Gly Arg Pro Leu Leu Met Val Phe Glu Tyr Met 580
585 590Arg His Gly Asp Leu Asn Arg Phe Leu Arg Ser
His Gly Pro Asp Ala 595 600 605Lys
Leu Leu Ala Gly Gly Glu Asp Val Ala Pro Gly Pro Leu Gly Leu 610
615 620Gly Gln Leu Leu Ala Val Ala Ser Gln Val
Ala Ala Gly Met Val Tyr625 630 635
640Leu Ala Gly Leu His Phe Val His Arg Asp Leu Ala Thr Arg Asn
Cys 645 650 655Leu Val Gly
Gln Gly Leu Val Val Lys Ile Gly Asp Phe Gly Met Ser 660
665 670Arg Asp Ile Tyr Ser Thr Asp Tyr Tyr Arg
Val Gly Gly Arg Thr Met 675 680
685Leu Pro Ile Arg Trp Met Pro Pro Glu Ser Ile Leu Tyr Arg Lys Phe 690
695 700Thr Thr Glu Ser Asp Val Trp Ser
Phe Gly Val Val Leu Trp Glu Ile705 710
715 720Phe Thr Tyr Gly Lys Gln Pro Trp Tyr Gln Leu Ser
Asn Thr Glu Ala 725 730
735Ile Asp Cys Ile Thr Gln Gly Arg Glu Leu Glu Arg Pro Arg Ala Cys
740 745 750Pro Pro Glu Val Tyr Ala
Ile Met Arg Gly Cys Trp Gln Arg Glu Pro 755 760
765Gln Gln Arg His Ser Ile Lys Asp Val His Ala Arg Leu Gln
Ala Leu 770 775 780Ala Gln Ala Pro Pro
Val Tyr Leu Asp Val Leu Gly785 790
79548569PRTHomo sapiens 48Met Lys Val Leu Leu Arg Leu Ile Cys Phe Ile Ala
Leu Leu Ile Ser1 5 10
15Ser Leu Glu Ala Asp Lys Cys Lys Glu Arg Glu Glu Lys Ile Ile Leu
20 25 30Val Ser Ser Ala Asn Glu Ile
Asp Val Arg Pro Cys Pro Leu Asn Pro 35 40
45Asn Glu His Lys Gly Thr Ile Thr Trp Tyr Lys Asp Asp Ser Lys
Thr 50 55 60Pro Val Ser Thr Glu Gln
Ala Ser Arg Ile His Gln His Lys Glu Lys65 70
75 80Leu Trp Phe Val Pro Ala Lys Val Glu Asp Ser
Gly His Tyr Tyr Cys 85 90
95Val Val Arg Asn Ser Ser Tyr Cys Leu Arg Ile Lys Ile Ser Ala Lys
100 105 110Phe Val Glu Asn Glu Pro
Asn Leu Cys Tyr Asn Ala Gln Ala Ile Phe 115 120
125Lys Gln Lys Leu Pro Val Ala Gly Asp Gly Gly Leu Val Cys
Pro Tyr 130 135 140Met Glu Phe Phe Lys
Asn Glu Asn Asn Glu Leu Pro Lys Leu Gln Trp145 150
155 160Tyr Lys Asp Cys Lys Pro Leu Leu Leu Asp
Asn Ile His Phe Ser Gly 165 170
175Val Lys Asp Arg Leu Ile Val Met Asn Val Ala Glu Lys His Arg Gly
180 185 190Asn Tyr Thr Cys His
Ala Ser Tyr Thr Tyr Leu Gly Lys Gln Tyr Pro 195
200 205Ile Thr Arg Val Ile Glu Phe Ile Thr Leu Glu Glu
Asn Lys Pro Thr 210 215 220Arg Pro Val
Ile Val Ser Pro Ala Asn Glu Thr Met Glu Val Asp Leu225
230 235 240Gly Ser Gln Ile Gln Leu Ile
Cys Asn Val Thr Gly Gln Leu Ser Asp 245
250 255Ile Ala Tyr Trp Lys Trp Asn Gly Ser Val Ile Asp
Glu Asp Asp Pro 260 265 270Val
Leu Gly Glu Asp Tyr Tyr Ser Val Glu Asn Pro Ala Asn Lys Arg 275
280 285Arg Ser Thr Leu Ile Thr Val Leu Asn
Ile Ser Glu Ile Glu Ser Arg 290 295
300Phe Tyr Lys His Pro Phe Thr Cys Phe Ala Lys Asn Thr His Gly Ile305
310 315 320Asp Ala Ala Tyr
Ile Gln Leu Ile Tyr Pro Val Thr Asn Phe Gln Lys 325
330 335His Met Ile Gly Ile Cys Val Thr Leu Thr
Val Ile Ile Val Cys Ser 340 345
350Val Phe Ile Tyr Lys Ile Phe Lys Ile Asp Ile Val Leu Trp Tyr Arg
355 360 365Asp Ser Cys Tyr Asp Phe Leu
Pro Ile Lys Ala Ser Asp Gly Lys Thr 370 375
380Tyr Asp Ala Tyr Ile Leu Tyr Pro Lys Thr Val Gly Glu Gly Ser
Thr385 390 395 400Ser Asp
Cys Asp Ile Phe Val Phe Lys Val Leu Pro Glu Val Leu Glu
405 410 415Lys Gln Cys Gly Tyr Lys Leu
Phe Ile Tyr Gly Arg Asp Asp Tyr Val 420 425
430Gly Glu Asp Ile Val Glu Val Ile Asn Glu Asn Val Lys Lys
Ser Arg 435 440 445Arg Leu Ile Ile
Ile Leu Val Arg Glu Thr Ser Gly Phe Ser Trp Leu 450
455 460Gly Gly Ser Ser Glu Glu Gln Ile Ala Met Tyr Asn
Ala Leu Val Gln465 470 475
480Asp Gly Ile Lys Val Val Leu Leu Glu Leu Glu Lys Ile Gln Asp Tyr
485 490 495Glu Lys Met Pro Glu
Ser Ile Lys Phe Ile Lys Gln Lys His Gly Ala 500
505 510Ile Arg Trp Ser Gly Asp Phe Thr Gln Gly Pro Gln
Ser Ala Lys Thr 515 520 525Arg Phe
Trp Lys Asn Val Arg Tyr His Met Pro Val Gln Arg Arg Ser 530
535 540Pro Ser Ser Lys His Gln Leu Leu Ser Pro Ala
Thr Lys Glu Lys Leu545 550 555
560Gln Arg Glu Ala His Val Pro Leu Gly
56549725PRTHomo sapiens 49Met Ala Asp Glu Asp Leu Ile Phe Arg Leu Glu Gly
Val Asp Gly Gly1 5 10
15Gln Ser Pro Arg Ala Gly His Asp Gly Asp Ser Asp Gly Asp Ser Asp
20 25 30Asp Glu Glu Gly Tyr Phe Ile
Cys Pro Ile Thr Asp Asp Pro Ser Ser 35 40
45Asn Gln Asn Val Asn Ser Lys Val Asn Lys Tyr Tyr Ser Asn Leu
Thr 50 55 60Lys Ser Glu Arg Tyr Ser
Ser Ser Gly Ser Pro Ala Asn Ser Phe His65 70
75 80Phe Lys Glu Ala Trp Lys His Ala Ile Gln Lys
Ala Lys His Met Pro 85 90
95Asp Pro Trp Ala Glu Phe His Leu Glu Asp Ile Ala Thr Glu Arg Ala
100 105 110Thr Arg His Arg Tyr Asn
Ala Val Thr Gly Glu Trp Leu Asp Asp Glu 115 120
125Val Leu Ile Lys Met Ala Ser Gln Pro Phe Gly Arg Gly Ala
Met Arg 130 135 140Glu Cys Phe Arg Thr
Lys Lys Leu Ser Asn Phe Leu His Ala Gln Gln145 150
155 160Trp Lys Gly Ala Ser Asn Tyr Val Ala Lys
Arg Tyr Ile Glu Pro Val 165 170
175Asp Arg Asp Val Tyr Phe Glu Asp Val Arg Leu Gln Met Glu Ala Lys
180 185 190Leu Trp Gly Glu Glu
Tyr Asn Arg His Lys Pro Pro Lys Gln Val Asp 195
200 205Ile Met Gln Met Cys Ile Ile Glu Leu Lys Asp Arg
Pro Gly Lys Pro 210 215 220Leu Phe His
Leu Glu His Tyr Ile Glu Gly Lys Tyr Ile Lys Tyr Asn225
230 235 240Ser Asn Ser Gly Phe Val Arg
Asp Asp Asn Ile Arg Leu Thr Pro Gln 245
250 255Ala Phe Ser His Phe Thr Phe Glu Arg Ser Gly His
Gln Leu Ile Val 260 265 270Val
Asp Ile Gln Gly Val Gly Asp Leu Tyr Thr Asp Pro Gln Ile His 275
280 285Thr Glu Thr Gly Thr Asp Phe Gly Asp
Gly Asn Leu Gly Val Arg Gly 290 295
300Met Ala Leu Phe Phe Tyr Ser His Ala Cys Asn Arg Ile Cys Glu Ser305
310 315 320Met Gly Leu Ala
Pro Phe Asp Leu Ser Pro Arg Glu Arg Asp Ala Val 325
330 335Asn Gln Asn Thr Lys Leu Leu Gln Ser Ala
Lys Thr Ile Leu Arg Gly 340 345
350Thr Glu Glu Lys Cys Gly Ser Pro Arg Val Arg Thr Leu Ser Gly Ser
355 360 365Arg Pro Pro Leu Leu Arg Pro
Leu Ser Glu Asn Ser Gly Asp Glu Asn 370 375
380Met Ser Asp Val Thr Phe Asp Ser Leu Pro Ser Ser Pro Ser Ser
Ala385 390 395 400Thr Pro
His Ser Gln Lys Leu Asp His Leu His Trp Pro Val Phe Ser
405 410 415Asp Leu Asp Asn Met Ala Ser
Arg Asp His Asp His Leu Asp Asn His 420 425
430Arg Glu Ser Glu Asn Ser Gly Asp Ser Gly Tyr Pro Ser Glu
Lys Arg 435 440 445Gly Glu Leu Asp
Asp Pro Glu Pro Arg Glu His Gly His Ser Tyr Ser 450
455 460Asn Arg Lys Tyr Glu Ser Asp Glu Asp Ser Leu Gly
Ser Ser Gly Arg465 470 475
480Val Cys Val Glu Lys Trp Asn Leu Leu Asn Ser Ser Arg Leu His Leu
485 490 495Pro Arg Ala Ser Ala
Val Ala Leu Glu Val Gln Arg Leu Asn Ala Leu 500
505 510Asp Leu Glu Lys Lys Ile Gly Lys Ser Ile Leu Gly
Lys Val His Leu 515 520 525Ala Met
Val Arg Tyr His Glu Gly Gly Arg Phe Cys Glu Lys Gly Glu 530
535 540Glu Trp Asp Gln Glu Ser Ala Val Phe His Leu
Glu His Ala Ala Asn545 550 555
560Leu Gly Glu Leu Glu Ala Ile Val Gly Leu Gly Leu Met Tyr Ser Gln
565 570 575Leu Pro His His
Ile Leu Ala Asp Val Ser Leu Lys Glu Thr Glu Glu 580
585 590Asn Lys Thr Lys Gly Phe Asp Tyr Leu Leu Lys
Ala Ala Glu Ala Gly 595 600 605Asp
Arg Gln Ser Met Ile Leu Val Ala Arg Ala Phe Asp Ser Gly Gln 610
615 620Asn Leu Ser Pro Asp Arg Cys Gln Asp Trp
Leu Glu Ala Leu His Trp625 630 635
640Tyr Asn Thr Ala Leu Glu Met Thr Asp Cys Asp Glu Gly Gly Glu
Tyr 645 650 655Asp Gly Met
Gln Asp Glu Pro Arg Tyr Met Met Leu Ala Arg Glu Ala 660
665 670Glu Met Leu Phe Thr Gly Gly Tyr Gly Leu
Glu Lys Asp Pro Gln Arg 675 680
685Ser Gly Asp Leu Tyr Thr Gln Ala Ala Glu Ala Ala Met Glu Ala Met 690
695 700Lys Gly Arg Leu Ala Asn Gln Tyr
Tyr Gln Lys Ala Glu Glu Ala Trp705 710
715 720Ala Gln Met Glu Glu 72550376PRTHomo
sapiens 50Met Cys Glu Glu Glu Thr Thr Ala Leu Val Cys Asp Asn Gly Ser
Gly1 5 10 15Leu Cys Lys
Ala Gly Phe Ala Gly Asp Asp Ala Pro Arg Ala Val Phe 20
25 30Pro Ser Ile Val Gly Arg Pro Arg His Gln
Gly Val Met Val Gly Met 35 40
45Gly Gln Lys Asp Ser Tyr Val Gly Asp Glu Ala Gln Ser Lys Arg Gly 50
55 60 Ile Leu Thr Leu Lys Tyr Pro Ile Glu
His Gly Ile Ile Thr Asn Trp65 70 75
80Asp Asp Met Glu Lys Ile Trp His His Ser Phe Tyr Asn Glu
Leu Arg 85 90 95Val Ala
Pro Glu Glu His Pro Thr Leu Leu Thr Glu Ala Pro Leu Asn 100
105 110Pro Lys Ala Asn Arg Glu Lys Met Thr
Gln Ile Met Phe Glu Thr Phe 115 120
125Asn Val Pro Ala Met Tyr Val Ala Ile Gln Ala Val Leu Ser Leu Tyr
130 135 140Ala Ser Gly Arg Thr Thr Gly
Ile Val Leu Asp Ser Gly Asp Gly Val145 150
155 160Thr His Asn Val Pro Ile Tyr Glu Gly Tyr Ala Leu
Pro His Ala Ile 165 170
175Met Arg Leu Asp Leu Ala Gly Arg Asp Leu Thr Asp Tyr Leu Met Lys
180 185 190Ile Leu Thr Glu Arg Gly
Tyr Ser Phe Val Thr Thr Ala Glu Arg Glu 195 200
205Ile Val Arg Asp Ile Lys Glu Lys Leu Cys Tyr Val Ala Leu
Asp Phe 210 215 220Glu Asn Glu Met Ala
Thr Ala Ala Ser Ser Ser Ser Leu Glu Lys Ser225 230
235 240Tyr Glu Leu Pro Asp Gly Gln Val Ile Thr
Ile Gly Asn Glu Arg Phe 245 250
255Arg Cys Pro Glu Thr Leu Phe Gln Pro Ser Phe Ile Gly Met Glu Ser
260 265 270Ala Gly Ile His Glu
Thr Thr Tyr Asn Ser Ile Met Lys Cys Asp Ile 275
280 285Asp Ile Arg Lys Asp Leu Tyr Ala Asn Asn Val Leu
Ser Gly Gly Thr 290 295 300Thr Met Tyr
Pro Gly Ile Ala Asp Arg Met Gln Lys Glu Ile Thr Ala305
310 315 320Leu Ala Pro Ser Thr Met Lys
Ile Lys Ile Ile Ala Pro Pro Glu Arg 325
330 335Lys Tyr Ser Val Trp Ile Gly Gly Ser Ile Leu Ala
Ser Leu Ser Thr 340 345 350Phe
Gln Gln Met Trp Ile Ser Lys Pro Glu Tyr Asp Glu Ala Gly Pro 355
360 365Ser Ile Val His Arg Lys Cys Phe
370 375511972PRTHomo sapiens 51Met Ala Gln Lys Gly Gln
Leu Ser Asp Asp Glu Lys Phe Leu Phe Val1 5
10 15Asp Lys Asn Phe Ile Asn Ser Pro Val Ala Gln Ala
Asp Trp Ala Ala 20 25 30Lys
Arg Leu Val Trp Val Pro Ser Glu Lys Gln Gly Phe Glu Ala Ala 35
40 45Ser Ile Lys Glu Glu Lys Gly Asp Glu
Val Val Val Glu Leu Val Glu 50 55
60Asn Gly Lys Lys Val Thr Val Gly Lys Asp Asp Ile Gln Lys Met Asn65
70 75 80Pro Pro Lys Phe Ser
Lys Val Glu Asp Met Ala Glu Leu Thr Cys Leu 85
90 95Asn Glu Ala Ser Val Leu His Asn Leu Arg Glu
Arg Tyr Phe Ser Gly 100 105
110Leu Ile Tyr Thr Tyr Ser Gly Leu Phe Cys Val Val Val Asn Pro Tyr
115 120 125Lys His Leu Pro Ile Tyr Ser
Glu Lys Ile Val Asp Met Tyr Lys Gly 130 135
140Lys Lys Arg His Glu Met Pro Pro His Ile Tyr Ala Ile Ala Asp
Thr145 150 155 160Ala Tyr
Arg Ser Met Leu Gln Asp Arg Glu Asp Gln Ser Ile Leu Cys
165 170 175Thr Gly Glu Ser Gly Ala Gly
Lys Thr Glu Asn Thr Lys Lys Val Ile 180 185
190Gln Tyr Leu Ala Val Val Ala Ser Ser His Lys Gly Lys Lys
Asp Thr 195 200 205Ser Ile Thr Gly
Glu Leu Glu Lys Gln Leu Leu Gln Ala Asn Pro Ile 210
215 220Leu Glu Ala Phe Gly Asn Ala Lys Thr Val Lys Asn
Asp Asn Ser Ser225 230 235
240Arg Phe Gly Lys Phe Ile Arg Ile Asn Phe Asp Val Thr Gly Tyr Ile
245 250 255Val Gly Ala Asn Ile
Glu Thr Tyr Leu Leu Glu Lys Ser Arg Ala Ile 260
265 270Arg Gln Ala Arg Asp Glu Arg Thr Phe His Ile Phe
Tyr Tyr Met Ile 275 280 285Ala Gly
Ala Lys Glu Lys Met Arg Ser Asp Leu Leu Leu Glu Gly Phe 290
295 300Asn Asn Tyr Thr Phe Leu Ser Asn Gly Phe Val
Pro Ile Pro Ala Ala305 310 315
320Gln Asp Asp Glu Met Phe Gln Glu Thr Val Glu Ala Met Ala Ile Met
325 330 335Gly Phe Ser Glu
Glu Glu Gln Leu Ser Ile Leu Lys Val Val Ser Ser 340
345 350Val Leu Gln Leu Gly Asn Ile Val Phe Lys Lys
Glu Arg Asn Thr Asp 355 360 365Gln
Ala Ser Met Pro Asp Asn Thr Ala Ala Gln Lys Val Cys His Leu 370
375 380Met Gly Ile Asn Val Thr Asp Phe Thr Arg
Ser Ile Leu Thr Pro Arg385 390 395
400Ile Lys Val Gly Arg Asp Val Val Gln Lys Ala Gln Thr Lys Glu
Gln 405 410 415Ala Asp Phe
Ala Val Glu Ala Leu Ala Lys Ala Thr Tyr Glu Arg Leu 420
425 430Phe Arg Trp Ile Leu Thr Arg Val Asn Lys
Ala Leu Asp Lys Thr His 435 440
445Arg Gln Gly Ala Ser Phe Leu Gly Ile Leu Asp Ile Ala Gly Phe Glu 450
455 460Ile Phe Glu Val Asn Ser Phe Glu
Gln Leu Cys Ile Asn Tyr Thr Asn465 470
475 480Glu Lys Leu Gln Gln Leu Phe Asn His Thr Met Phe
Ile Leu Glu Gln 485 490
495Glu Glu Tyr Gln Arg Glu Gly Ile Glu Trp Asn Phe Ile Asp Phe Gly
500 505 510Leu Asp Leu Gln Pro Cys
Ile Glu Leu Ile Glu Arg Pro Asn Asn Pro 515 520
525Pro Gly Val Leu Ala Leu Leu Asp Glu Glu Cys Trp Phe Pro
Lys Ala 530 535 540Thr Asp Lys Ser Phe
Val Glu Lys Leu Cys Thr Glu Gln Gly Ser His545 550
555 560Pro Lys Phe Gln Lys Pro Lys Gln Leu Lys
Asp Lys Thr Glu Phe Ser 565 570
575Ile Ile His Tyr Ala Gly Lys Val Asp Tyr Asn Ala Ser Ala Trp Leu
580 585 590Thr Lys Asn Met Asp
Pro Leu Asn Asp Asn Val Thr Ser Leu Leu Asn 595
600 605Ala Ser Ser Asp Lys Phe Val Ala Asp Leu Trp Lys
Asp Val Asp Arg 610 615 620Ile Val Gly
Leu Asp Gln Met Ala Lys Met Thr Glu Ser Ser Leu Pro625
630 635 640Ser Ala Ser Lys Thr Lys Lys
Gly Met Phe Arg Thr Val Gly Gln Leu 645
650 655Tyr Lys Glu Gln Leu Gly Lys Leu Met Thr Thr Leu
Arg Asn Thr Thr 660 665 670Pro
Asn Phe Val Arg Cys Ile Ile Pro Asn His Glu Lys Arg Ser Gly 675
680 685Lys Leu Asp Ala Phe Leu Val Leu Glu
Gln Leu Arg Cys Asn Gly Val 690 695
700Leu Glu Gly Ile Arg Ile Cys Arg Gln Gly Phe Pro Asn Arg Ile Val705
710 715 720Phe Gln Glu Phe
Arg Gln Arg Tyr Glu Ile Leu Ala Ala Asn Ala Ile 725
730 735Pro Lys Gly Phe Met Asp Gly Lys Gln Ala
Cys Ile Leu Met Ile Lys 740 745
750Ala Leu Glu Leu Asp Pro Asn Leu Tyr Arg Ile Gly Gln Ser Lys Ile
755 760 765Phe Phe Arg Thr Gly Val Leu
Ala His Leu Glu Glu Glu Arg Asp Leu 770 775
780Lys Ile Thr Asp Val Ile Met Ala Phe Gln Ala Met Cys Arg Gly
Tyr785 790 795 800Leu Ala
Arg Lys Ala Phe Ala Lys Arg Gln Gln Gln Leu Thr Ala Met
805 810 815Lys Val Ile Gln Arg Asn Cys
Ala Ala Tyr Leu Lys Leu Arg Asn Trp 820 825
830Gln Trp Trp Arg Leu Phe Thr Lys Val Lys Pro Leu Leu Gln
Val Thr 835 840 845Arg Gln Glu Glu
Glu Met Gln Ala Lys Glu Asp Glu Leu Gln Lys Thr 850
855 860Lys Glu Arg Gln Gln Lys Ala Glu Asn Glu Leu Lys
Glu Leu Glu Gln865 870 875
880Lys His Ser Gln Leu Thr Glu Glu Lys Asn Leu Leu Gln Glu Gln Leu
885 890 895Gln Ala Glu Thr Glu
Leu Tyr Ala Glu Ala Glu Glu Met Arg Val Arg 900
905 910Leu Ala Ala Lys Lys Gln Glu Leu Glu Glu Ile Leu
His Glu Met Glu 915 920 925Ala Arg
Leu Glu Glu Glu Glu Asp Arg Gly Gln Gln Leu Gln Ala Glu 930
935 940Arg Lys Lys Met Ala Gln Gln Met Leu Asp Leu
Glu Glu Gln Leu Glu945 950 955
960Glu Glu Glu Ala Ala Arg Gln Lys Leu Gln Leu Glu Lys Val Thr Ala
965 970 975Glu Ala Lys Ile
Lys Lys Leu Glu Asp Glu Ile Leu Val Met Asp Asp 980
985 990Gln Asn Asn Lys Leu Ser Lys Glu Arg Lys Leu
Leu Glu Glu Arg Ile 995 1000
1005Ser Asp Leu Thr Thr Asn Leu Ala Glu Glu Glu Glu Lys Ala Lys
1010 1015 1020Asn Leu Thr Lys Leu Lys
Asn Lys His Glu Ser Met Ile Ser Glu 1025 1030
1035Leu Glu Val Arg Leu Lys Lys Glu Glu Lys Ser Arg Gln Glu
Leu 1040 1045 1050Glu Lys Leu Lys Arg
Lys Leu Glu Gly Asp Ala Ser Asp Phe His 1055 1060
1065Glu Gln Ile Ala Asp Leu Gln Ala Gln Ile Ala Glu Leu
Lys Met 1070 1075 1080Gln Leu Ala Lys
Lys Glu Glu Glu Leu Gln Ala Ala Leu Ala Arg 1085
1090 1095Leu Asp Asp Glu Ile Ala Gln Lys Asn Asn Ala
Leu Lys Lys Ile 1100 1105 1110Arg Glu
Leu Glu Gly His Ile Ser Asp Leu Gln Glu Asp Leu Asp 1115
1120 1125Ser Glu Arg Ala Ala Arg Asn Lys Ala Glu
Lys Gln Lys Arg Asp 1130 1135 1140Leu
Gly Glu Glu Leu Glu Ala Leu Lys Thr Glu Leu Glu Asp Thr 1145
1150 1155Leu Asp Ser Thr Ala Thr Gln Gln Glu
Leu Arg Ala Lys Arg Glu 1160 1165
1170Gln Glu Val Thr Val Leu Lys Lys Ala Leu Asp Glu Glu Thr Arg
1175 1180 1185Ser His Glu Ala Gln Val
Gln Glu Met Arg Gln Lys His Ala Gln 1190 1195
1200Ala Val Glu Glu Leu Thr Glu Gln Leu Glu Gln Phe Lys Arg
Ala 1205 1210 1215Lys Ala Asn Leu Asp
Lys Asn Lys Gln Thr Leu Glu Lys Glu Asn 1220 1225
1230Ala Asp Leu Ala Gly Glu Leu Arg Val Leu Gly Gln Ala
Lys Gln 1235 1240 1245Glu Val Glu His
Lys Lys Lys Lys Leu Glu Ala Gln Val Gln Glu 1250
1255 1260Leu Gln Ser Lys Cys Ser Asp Gly Glu Arg Ala
Arg Ala Glu Leu 1265 1270 1275Asn Asp
Lys Val His Lys Leu Gln Asn Glu Val Glu Ser Val Thr 1280
1285 1290Gly Met Leu Asn Glu Ala Glu Gly Lys Ala
Ile Lys Leu Ala Lys 1295 1300 1305Asp
Val Ala Ser Leu Ser Ser Gln Leu Gln Asp Thr Gln Glu Leu 1310
1315 1320Leu Gln Glu Glu Thr Arg Gln Lys Leu
Asn Val Ser Thr Lys Leu 1325 1330
1335Arg Gln Leu Glu Glu Glu Arg Asn Ser Leu Gln Asp Gln Leu Asp
1340 1345 1350Glu Glu Met Glu Ala Lys
Gln Asn Leu Glu Arg His Ile Ser Thr 1355 1360
1365Leu Asn Ile Gln Leu Ser Asp Ser Lys Lys Lys Leu Gln Asp
Phe 1370 1375 1380Ala Ser Thr Val Glu
Ala Leu Glu Glu Gly Lys Lys Arg Phe Gln 1385 1390
1395Lys Glu Ile Glu Asn Leu Thr Gln Gln Tyr Glu Glu Lys
Ala Ala 1400 1405 1410Ala Tyr Asp Lys
Leu Glu Lys Thr Lys Asn Arg Leu Gln Gln Glu 1415
1420 1425Leu Asp Asp Leu Val Val Asp Leu Asp Asn Gln
Arg Gln Leu Val 1430 1435 1440Ser Asn
Leu Glu Lys Lys Gln Arg Lys Phe Asp Gln Leu Leu Ala 1445
1450 1455Glu Glu Lys Asn Ile Ser Ser Lys Tyr Ala
Asp Glu Arg Asp Arg 1460 1465 1470Ala
Glu Ala Glu Ala Arg Glu Lys Glu Thr Lys Ala Leu Ser Leu 1475
1480 1485Ala Arg Ala Leu Glu Glu Ala Leu Glu
Ala Lys Glu Glu Leu Glu 1490 1495
1500Arg Thr Asn Lys Met Leu Lys Ala Glu Met Glu Asp Leu Val Ser
1505 1510 1515Ser Lys Asp Asp Val Gly
Lys Asn Val His Glu Leu Glu Lys Ser 1520 1525
1530Lys Arg Ala Leu Glu Thr Gln Met Glu Glu Met Lys Thr Gln
Leu 1535 1540 1545Glu Glu Leu Glu Asp
Glu Leu Gln Ala Thr Glu Asp Ala Lys Leu 1550 1555
1560Arg Leu Glu Val Asn Met Gln Ala Leu Lys Gly Gln Phe
Glu Arg 1565 1570 1575Asp Leu Gln Ala
Arg Asp Glu Gln Asn Glu Glu Lys Arg Arg Gln 1580
1585 1590Leu Gln Arg Gln Leu His Glu Tyr Glu Thr Glu
Leu Glu Asp Glu 1595 1600 1605Arg Lys
Gln Arg Ala Leu Ala Ala Ala Ala Lys Lys Lys Leu Glu 1610
1615 1620Gly Asp Leu Lys Asp Leu Glu Leu Gln Ala
Asp Ser Ala Ile Lys 1625 1630 1635Gly
Arg Glu Glu Ala Ile Lys Gln Leu Arg Lys Leu Gln Ala Gln 1640
1645 1650Met Lys Asp Phe Gln Arg Glu Leu Glu
Asp Ala Arg Ala Ser Arg 1655 1660
1665Asp Glu Ile Phe Ala Thr Ala Lys Glu Asn Glu Lys Lys Ala Lys
1670 1675 1680Ser Leu Glu Ala Asp Leu
Met Gln Leu Gln Glu Asp Leu Ala Ala 1685 1690
1695Ala Glu Arg Ala Arg Lys Gln Ala Asp Leu Glu Lys Glu Glu
Leu 1700 1705 1710Ala Glu Glu Leu Ala
Ser Ser Leu Ser Gly Arg Asn Ala Leu Gln 1715 1720
1725Asp Glu Lys Arg Arg Leu Glu Ala Arg Ile Ala Gln Leu
Glu Glu 1730 1735 1740Glu Leu Glu Glu
Glu Gln Gly Asn Met Glu Ala Met Ser Asp Arg 1745
1750 1755Val Arg Lys Ala Thr Gln Gln Ala Glu Gln Leu
Ser Asn Glu Leu 1760 1765 1770Ala Thr
Glu Arg Ser Thr Ala Gln Lys Asn Glu Ser Ala Arg Gln 1775
1780 1785Gln Leu Glu Arg Gln Asn Lys Glu Leu Arg
Ser Lys Leu His Glu 1790 1795 1800Met
Glu Gly Ala Val Lys Ser Lys Phe Lys Ser Thr Ile Ala Ala 1805
1810 1815Leu Glu Ala Lys Ile Ala Gln Leu Glu
Glu Gln Val Glu Gln Glu 1820 1825
1830Ala Arg Glu Lys Gln Ala Ala Thr Lys Ser Leu Lys Gln Lys Asp
1835 1840 1845Lys Lys Leu Lys Glu Ile
Leu Leu Gln Val Glu Asp Glu Arg Lys 1850 1855
1860Met Ala Glu Gln Tyr Lys Glu Gln Ala Glu Lys Gly Asn Ala
Arg 1865 1870 1875Val Lys Gln Leu Lys
Arg Gln Leu Glu Glu Ala Glu Glu Glu Ser 1880 1885
1890Gln Arg Ile Asn Ala Asn Arg Arg Lys Leu Gln Arg Glu
Leu Asp 1895 1900 1905Glu Ala Thr Glu
Ser Asn Glu Ala Met Gly Arg Glu Val Asn Ala 1910
1915 1920Leu Lys Ser Lys Leu Arg Arg Gly Asn Glu Thr
Ser Phe Val Pro 1925 1930 1935Ser Arg
Arg Ser Gly Gly Arg Arg Val Ile Glu Asn Ala Asp Gly 1940
1945 1950Ser Glu Glu Glu Thr Asp Thr Arg Asp Ala
Asp Phe Asn Gly Thr 1955 1960 1965Lys
Ala Ser Glu 197052171PRTHomo sapiens 52Met Ser Ser Lys Arg Thr Lys
Thr Lys Thr Lys Lys Arg Pro Gln Arg1 5 10
15Ala Thr Ser Asn Val Phe Ala Met Phe Asp Gln Ser Gln
Ile Gln Glu 20 25 30Phe Lys
Glu Ala Phe Asn Met Ile Asp Gln Asn Arg Asp Gly Phe Ile 35
40 45Asp Lys Glu Asp Leu His Asp Met Leu Ala
Ser Leu Gly Lys Asn Pro 50 55 60Thr
Asp Glu Tyr Leu Asp Ala Met Met Asn Glu Ala Pro Gly Pro Ile65
70 75 80Asn Phe Thr Met Phe Leu
Thr Met Phe Gly Glu Lys Leu Asn Gly Thr 85
90 95Asp Pro Glu Asp Val Ile Arg Asn Ala Phe Ala Cys
Phe Asp Glu Glu 100 105 110Ala
Thr Gly Thr Ile Gln Glu Asp Tyr Leu Arg Glu Leu Leu Thr Thr 115
120 125Met Gly Asp Arg Phe Thr Asp Glu Glu
Val Asp Glu Leu Tyr Arg Glu 130 135
140Ala Pro Ile Asp Lys Lys Gly Asn Phe Asn Tyr Ile Glu Phe Thr Arg145
150 155 160Ile Leu Lys His
Gly Ala Lys Asp Lys Asp Asp 165
17053172PRTHomo sapiens 53Met Ser Ser Lys Lys Ala Lys Thr Lys Thr Thr Lys
Lys Arg Pro Gln1 5 10
15Arg Ala Thr Ser Asn Val Phe Ala Met Phe Asp Gln Ser Gln Ile Gln
20 25 30Glu Phe Lys Glu Ala Phe Asn
Met Ile Asp Gln Asn Arg Asp Gly Phe 35 40
45Ile Asp Lys Glu Asp Leu His Asp Met Leu Ala Ser Leu Gly Lys
Asn 50 55 60Pro Thr Asp Ala Tyr Leu
Asp Ala Met Met Asn Glu Ala Pro Gly Pro65 70
75 80Ile Asn Phe Thr Met Phe Leu Thr Met Phe Gly
Glu Lys Leu Asn Gly 85 90
95Thr Asp Pro Glu Asp Val Ile Arg Asn Ala Phe Ala Cys Phe Asp Glu
100 105 110Glu Ala Thr Gly Thr Ile
Gln Glu Asp Tyr Leu Arg Glu Leu Leu Thr 115 120
125Thr Met Gly Asp Arg Phe Thr Asp Glu Glu Val Asp Glu Leu
Tyr Arg 130 135 140Glu Ala Pro Ile Asp
Lys Lys Gly Asn Phe Asn Tyr Ile Glu Phe Thr145 150
155 160Arg Ile Leu Lys His Gly Ala Lys Asp Lys
Asp Asp 165 17054232PRTHomo sapiens 54Met
Lys Glu Arg Arg Ala Pro Gln Pro Val Val Ala Arg Cys Lys Leu1
5 10 15Val Leu Val Gly Asp Val Gln
Cys Gly Lys Thr Ala Met Leu Gln Val 20 25
30Leu Ala Lys Asp Cys Tyr Pro Glu Thr Tyr Val Pro Thr Val
Phe Glu 35 40 45Asn Tyr Thr Ala
Cys Leu Glu Thr Glu Glu Gln Arg Val Glu Leu Ser 50 55
60Leu Trp Asp Thr Ser Gly Ser Pro Tyr Tyr Asp Asn Val
Arg Pro Leu65 70 75
80Cys Tyr Ser Asp Ser Asp Ala Val Leu Leu Cys Phe Asp Ile Ser Arg
85 90 95Pro Glu Thr Val Asp Ser
Ala Leu Lys Lys Trp Arg Thr Glu Ile Leu 100
105 110Asp Tyr Cys Pro Ser Thr Arg Val Leu Leu Ile Gly
Cys Lys Thr Asp 115 120 125Leu Arg
Thr Asp Leu Ser Thr Leu Met Glu Leu Ser His Gln Lys Gln 130
135 140Ala Pro Ile Ser Tyr Glu Gln Gly Cys Ala Ile
Ala Lys Gln Leu Gly145 150 155
160Ala Glu Ile Tyr Leu Glu Gly Ser Ala Phe Thr Ser Glu Lys Ser Ile
165 170 175His Ser Ile Phe
Arg Thr Ala Ser Met Leu Cys Leu Asn Lys Pro Ser 180
185 190Pro Leu Pro Gln Lys Ser Pro Val Arg Ser Leu
Ser Lys Arg Leu Leu 195 200 205His
Leu Pro Ser Arg Ser Glu Leu Ile Ser Ser Thr Phe Lys Lys Glu 210
215 220Lys Ala Lys Ser Cys Ser Ile Met225
23055205PRTHomo sapiens 55Met Thr Glu Arg Arg Val Pro Phe Ser
Leu Leu Arg Gly Pro Ser Trp1 5 10
15Asp Pro Phe Arg Asp Trp Tyr Pro His Ser Arg Leu Phe Asp Gln
Ala 20 25 30Phe Gly Leu Pro
Arg Leu Pro Glu Glu Trp Ser Gln Trp Leu Gly Gly 35
40 45Ser Ser Trp Pro Gly Tyr Val Arg Pro Leu Pro Pro
Ala Ala Ile Glu 50 55 60Ser Pro Ala
Val Ala Ala Pro Ala Tyr Ser Arg Ala Leu Ser Arg Gln65 70
75 80Leu Ser Ser Gly Val Ser Glu Ile
Arg His Thr Ala Asp Arg Trp Arg 85 90
95Val Ser Leu Asp Val Asn His Phe Ala Pro Asp Glu Leu Thr
Val Lys 100 105 110Thr Lys Asp
Gly Val Val Glu Ile Thr Gly Lys His Glu Glu Arg Gln 115
120 125Asp Glu His Gly Tyr Ile Ser Arg Cys Phe Thr
Arg Lys Tyr Thr Leu 130 135 140Pro Pro
Gly Val Asp Pro Thr Gln Val Ser Ser Ser Leu Ser Pro Glu145
150 155 160Gly Thr Leu Thr Val Glu Ala
Pro Met Pro Lys Leu Ala Thr Gln Ser 165
170 175Asn Glu Ile Thr Ile Pro Val Thr Phe Glu Ser Arg
Ala Gln Leu Gly 180 185 190Gly
Pro Glu Ala Ala Lys Ser Asp Glu Thr Ala Ala Lys 195
200 20556784PRTHomo sapiens 56Met Glu Gly Asp Gly Gly
Thr Pro Trp Ala Leu Ala Leu Leu Arg Thr1 5
10 15Phe Asp Ala Gly Glu Phe Thr Gly Trp Glu Lys Val
Gly Ser Gly Gly 20 25 30Phe
Gly Gln Val Tyr Lys Val Arg His Val His Trp Lys Thr Trp Leu 35
40 45Ala Ile Lys Cys Ser Pro Ser Leu His
Val Asp Asp Arg Glu Arg Met 50 55
60Glu Leu Leu Glu Glu Ala Lys Lys Met Glu Met Ala Lys Phe Arg Tyr65
70 75 80Ile Leu Pro Val Tyr
Gly Ile Cys Arg Glu Pro Val Gly Leu Val Met 85
90 95Glu Tyr Met Glu Thr Gly Ser Leu Glu Lys Leu
Leu Ala Ser Glu Pro 100 105
110Leu Pro Trp Asp Leu Arg Phe Arg Ile Ile His Glu Thr Ala Val Gly
115 120 125Met Asn Phe Leu His Cys Met
Ala Pro Pro Leu Leu His Leu Asp Leu 130 135
140Lys Pro Ala Asn Ile Leu Leu Asp Ala His Tyr His Val Lys Ile
Ser145 150 155 160Asp Phe
Gly Leu Ala Lys Cys Asn Gly Leu Ser His Ser His Asp Leu
165 170 175Ser Met Asp Gly Leu Phe Gly
Thr Ile Ala Tyr Leu Pro Pro Glu Arg 180 185
190Ile Arg Glu Lys Ser Arg Leu Phe Asp Thr Lys His Asp Val
Tyr Ser 195 200 205Phe Ala Ile Val
Ile Trp Gly Val Leu Thr Gln Lys Lys Pro Phe Ala 210
215 220Asp Glu Lys Asn Ile Leu His Ile Met Val Lys Val
Val Lys Gly His225 230 235
240Arg Pro Glu Leu Pro Pro Val Cys Arg Ala Arg Pro Arg Ala Cys Ser
245 250 255His Leu Ile Arg Leu
Met Gln Arg Cys Trp Gln Gly Asp Pro Arg Val 260
265 270Arg Pro Thr Phe Gln Glu Ile Thr Ser Glu Thr Glu
Asp Leu Cys Glu 275 280 285Lys Pro
Asp Asp Glu Val Lys Glu Thr Ala His Asp Leu Asp Val Lys 290
295 300Ser Pro Pro Glu Pro Arg Ser Glu Val Val Pro
Ala Arg Leu Lys Arg305 310 315
320Ala Ser Ala Pro Thr Phe Asp Asn Asp Tyr Ser Leu Ser Glu Leu Leu
325 330 335Ser Gln Leu Asp
Ser Gly Val Ser Gln Ala Val Glu Gly Pro Glu Glu 340
345 350Leu Ser Arg Ser Ser Ser Glu Ser Lys Leu Pro
Ser Ser Gly Ser Gly 355 360 365Lys
Arg Leu Ser Gly Val Ser Ser Val Asp Ser Ala Phe Ser Ser Arg 370
375 380Gly Ser Leu Ser Leu Ser Phe Glu Arg Glu
Pro Ser Thr Ser Asp Leu385 390 395
400Gly Thr Thr Asp Val Gln Lys Lys Lys Leu Val Asp Ala Ile Val
Ser 405 410 415Gly Asp Thr
Ser Lys Leu Met Lys Ile Leu Gln Pro Gln Asp Val Asp 420
425 430Leu Ala Leu Asp Ser Gly Ala Ser Leu Leu
His Leu Ala Val Glu Ala 435 440
445Gly Gln Glu Glu Cys Ala Lys Trp Leu Leu Leu Asn Asn Ala Asn Pro 450
455 460Asn Leu Ser Asn Arg Arg Gly Ser
Thr Pro Leu His Met Ala Val Glu465 470
475 480Arg Arg Val Arg Gly Val Val Glu Leu Leu Leu Ala
Arg Lys Ile Ser 485 490
495Val Asn Ala Lys Asp Glu Asp Gln Trp Thr Ala Leu His Phe Ala Ala
500 505 510Gln Asn Gly Asp Glu Ser
Ser Thr Arg Leu Leu Leu Glu Lys Asn Ala 515 520
525Ser Val Asn Glu Val Asp Phe Glu Gly Arg Thr Pro Met His
Val Ala 530 535 540Cys Gln His Gly Gln
Glu Asn Ile Val Arg Ile Leu Leu Arg Arg Gly545 550
555 560Val Asp Val Ser Leu Gln Gly Lys Asp Ala
Trp Leu Pro Leu His Tyr 565 570
575Ala Ala Trp Gln Gly His Leu Pro Ile Val Lys Leu Leu Ala Lys Gln
580 585 590Pro Gly Val Ser Val
Asn Ala Gln Thr Leu Asp Gly Arg Thr Pro Leu 595
600 605His Leu Ala Ala Gln Arg Gly His Tyr Arg Val Ala
Arg Ile Leu Ile 610 615 620Asp Leu Cys
Ser Asp Val Asn Val Cys Ser Leu Leu Ala Gln Thr Pro625
630 635 640Leu His Val Ala Ala Glu Thr
Gly His Thr Ser Thr Ala Arg Leu Leu 645
650 655Leu His Arg Gly Ala Gly Lys Glu Ala Met Thr Ser
Asp Gly Tyr Thr 660 665 670Ala
Leu His Leu Ala Ala Arg Asn Gly His Leu Ala Thr Val Lys Leu 675
680 685Leu Val Glu Glu Lys Ala Asp Val Leu
Ala Arg Gly Pro Leu Asn Gln 690 695
700Thr Ala Leu His Leu Ala Ala Ala His Gly His Ser Glu Val Val Glu705
710 715 720Glu Leu Val Ser
Ala Asp Val Ile Asp Leu Phe Asp Glu Gln Gly Leu 725
730 735Ser Ala Leu His Leu Ala Ala Gln Gly Arg
His Ala Gln Thr Val Glu 740 745
750Thr Leu Leu Arg His Gly Ala His Ile Asn Leu Gln Ser Leu Lys Phe
755 760 765Gln Gly Gly His Gly Pro Ala
Ala Thr Leu Leu Arg Arg Ser Lys Thr 770 775
78057202PRTHomo sapiens 57Met Ser Ala Ser Thr Ser Ser His Arg Pro
Ile Lys Gly Ile Leu Lys1 5 10
15Asn Lys Ser Ser Ser Gly Ser Ser Val Ala Thr Ser Gly Gln Gln Ser
20 25 30Gly Gly Thr Ile Gln Asp
Val Lys Arg Lys Lys Ser Gln Lys Trp Asp 35 40
45Glu Ser Ser Ile Leu Ala Ala His Arg Ala Thr Tyr Arg Asp
Tyr Asp 50 55 60Leu Met Lys Ala Asn
Glu Pro Gly Thr Ser Tyr Met Ser Val Gln Asp65 70
75 80Asn Gly Glu Asp Ser Val Arg Asp Val Glu
Gly Glu Asp Ser Val Arg 85 90
95Gly Val Glu Gly Lys Glu Ala Thr Asp Ala Ser Asp His Ser Cys Glu
100 105 110Val Asp Glu Gln Glu
Ser Ser Glu Ala Tyr Met Arg Lys Ile Leu Leu 115
120 125His Lys Gln Glu Lys Lys Arg Gln Phe Glu Met Arg
Arg Arg Leu His 130 135 140Tyr Asn Glu
Glu Leu Asn Ile Lys Leu Ala Arg Gln Leu Met Trp Lys145
150 155 160Glu Leu Gln Ser Glu Asp Asn
Glu Asn Glu Glu Thr Pro Gln Gly Thr 165
170 175Asn Glu Glu Lys Thr Ala Ala Glu Glu Ser Glu Glu
Ala Pro Leu Thr 180 185 190Gly
Gly Leu Gln Thr Gln Ser Cys Asp Pro 195
20058201PRTHomo sapiens 58Met Ala Asn Lys Gly Pro Ser Tyr Gly Met Ser Arg
Glu Val Gln Ser1 5 10
15Lys Ile Glu Lys Lys Tyr Asp Glu Glu Leu Glu Glu Arg Leu Val Glu
20 25 30Trp Ile Ile Val Gln Cys Gly
Pro Asp Val Gly Arg Pro Asp Arg Gly 35 40
45Arg Leu Gly Phe Gln Val Trp Leu Lys Asn Gly Val Ile Leu Ser
Lys 50 55 60Leu Val Asn Ser Leu Tyr
Pro Asp Gly Ser Lys Pro Val Lys Val Pro65 70
75 80Glu Asn Pro Pro Ser Met Val Phe Lys Gln Met
Glu Gln Val Ala Gln 85 90
95Phe Leu Lys Ala Ala Glu Asp Tyr Gly Val Ile Lys Thr Asp Met Phe
100 105 110Gln Thr Val Asp Leu Phe
Glu Gly Lys Asp Met Ala Ala Val Gln Arg 115 120
125Thr Leu Met Ala Leu Gly Ser Leu Ala Val Thr Lys Asn Asp
Gly His 130 135 140Tyr Arg Gly Asp Pro
Asn Trp Phe Met Lys Lys Ala Gln Glu His Lys145 150
155 160Arg Glu Phe Thr Glu Ser Gln Leu Gln Glu
Gly Lys His Val Ile Gly 165 170
175Leu Gln Met Gly Ser Asn Arg Gly Ala Ser Gln Ala Gly Met Thr Gly
180 185 190Tyr Gly Arg Pro Arg
Gln Ile Ile Ser 195 20059798PRTHomo sapiens 59Met
Asn Leu Gln Pro Ile Phe Trp Ile Gly Leu Ile Ser Ser Val Cys1
5 10 15Cys Val Phe Ala Gln Thr Asp
Glu Asn Arg Cys Leu Lys Ala Asn Ala 20 25
30Lys Ser Cys Gly Glu Cys Ile Gln Ala Gly Pro Asn Cys Gly
Trp Cys 35 40 45Thr Asn Ser Thr
Phe Leu Gln Glu Gly Met Pro Thr Ser Ala Arg Cys 50 55
60Asp Asp Leu Glu Ala Leu Lys Lys Lys Gly Cys Pro Pro
Asp Asp Ile65 70 75
80Glu Asn Pro Arg Gly Ser Lys Asp Ile Lys Lys Asn Lys Asn Val Thr
85 90 95Asn Arg Ser Lys Gly Thr
Ala Glu Lys Leu Lys Pro Glu Asp Ile Thr 100
105 110Gln Ile Gln Pro Gln Gln Leu Val Leu Arg Leu Arg
Ser Gly Glu Pro 115 120 125Gln Thr
Phe Thr Leu Lys Phe Lys Arg Ala Glu Asp Tyr Pro Ile Asp 130
135 140Leu Tyr Tyr Leu Met Asp Leu Ser Tyr Ser Met
Lys Asp Asp Leu Glu145 150 155
160Asn Val Lys Ser Leu Gly Thr Asp Leu Met Asn Glu Met Arg Arg Ile
165 170 175Thr Ser Asp Phe
Arg Ile Gly Phe Gly Ser Phe Val Glu Lys Thr Val 180
185 190Met Pro Tyr Ile Ser Thr Thr Pro Ala Lys Leu
Arg Asn Pro Cys Thr 195 200 205Ser
Glu Gln Asn Cys Thr Ser Pro Phe Ser Tyr Lys Asn Val Leu Ser 210
215 220Leu Thr Asn Lys Gly Glu Val Phe Asn Glu
Leu Val Gly Lys Gln Arg225 230 235
240Ile Ser Gly Asn Leu Asp Ser Pro Glu Gly Gly Phe Asp Ala Ile
Met 245 250 255Gln Val Ala
Val Cys Gly Ser Leu Ile Gly Trp Arg Asn Val Thr Arg 260
265 270Leu Leu Val Phe Ser Thr Asp Ala Gly Phe
His Phe Ala Gly Asp Gly 275 280
285Lys Leu Gly Gly Ile Val Leu Pro Asn Asp Gly Gln Cys His Leu Glu 290
295 300Asn Asn Met Tyr Thr Met Ser His
Tyr Tyr Asp Tyr Pro Ser Ile Ala305 310
315 320His Leu Val Gln Lys Leu Ser Glu Asn Asn Ile Gln
Thr Ile Phe Ala 325 330
335Val Thr Glu Glu Phe Gln Pro Val Tyr Lys Glu Leu Lys Asn Leu Ile
340 345 350Pro Lys Ser Ala Val Gly
Thr Leu Ser Ala Asn Ser Ser Asn Val Ile 355 360
365Gln Leu Ile Ile Asp Ala Tyr Asn Ser Leu Ser Ser Glu Val
Ile Leu 370 375 380Glu Asn Gly Lys Leu
Ser Glu Gly Val Thr Ile Ser Tyr Lys Ser Tyr385 390
395 400Cys Lys Asn Gly Val Asn Gly Thr Gly Glu
Asn Gly Arg Lys Cys Ser 405 410
415Asn Ile Ser Ile Gly Asp Glu Val Gln Phe Glu Ile Ser Ile Thr Ser
420 425 430Asn Lys Cys Pro Lys
Lys Asp Ser Asp Ser Phe Lys Ile Arg Pro Leu 435
440 445Gly Phe Thr Glu Glu Val Glu Val Ile Leu Gln Tyr
Ile Cys Glu Cys 450 455 460Glu Cys Gln
Ser Glu Gly Ile Pro Glu Ser Pro Lys Cys His Glu Gly465
470 475 480Asn Gly Thr Phe Glu Cys Gly
Ala Cys Arg Cys Asn Glu Gly Arg Val 485
490 495Gly Arg His Cys Glu Cys Ser Thr Asp Glu Val Asn
Ser Glu Asp Met 500 505 510Asp
Ala Tyr Cys Arg Lys Glu Asn Ser Ser Glu Ile Cys Ser Asn Asn 515
520 525Gly Glu Cys Val Cys Gly Gln Cys Val
Cys Arg Lys Arg Asp Asn Thr 530 535
540Asn Glu Ile Tyr Ser Gly Lys Phe Cys Glu Cys Asp Asn Phe Asn Cys545
550 555 560Asp Arg Ser Asn
Gly Leu Ile Cys Gly Gly Asn Gly Val Cys Lys Cys 565
570 575Arg Val Cys Glu Cys Asn Pro Asn Tyr Thr
Gly Ser Ala Cys Asp Cys 580 585
590Ser Leu Asp Thr Ser Thr Cys Glu Ala Ser Asn Gly Gln Ile Cys Asn
595 600 605Gly Arg Gly Ile Cys Glu Cys
Gly Val Cys Lys Cys Thr Asp Pro Lys 610 615
620Phe Gln Gly Gln Thr Cys Glu Met Cys Gln Thr Cys Leu Gly Val
Cys625 630 635 640Ala Glu
His Lys Glu Cys Val Gln Cys Arg Ala Phe Asn Lys Gly Glu
645 650 655Lys Lys Asp Thr Cys Thr Gln
Glu Cys Ser Tyr Phe Asn Ile Thr Lys 660 665
670Val Glu Ser Arg Asp Lys Leu Pro Gln Pro Val Gln Pro Asp
Pro Val 675 680 685Ser His Cys Lys
Glu Lys Asp Val Asp Asp Cys Trp Phe Tyr Phe Thr 690
695 700Tyr Ser Val Asn Gly Asn Asn Glu Val Met Val His
Val Val Glu Asn705 710 715
720Pro Glu Cys Pro Thr Gly Pro Asp Ile Ile Pro Ile Val Ala Gly Val
725 730 735Val Ala Gly Ile Val
Leu Ile Gly Leu Ala Leu Leu Leu Ile Trp Lys 740
745 750Leu Leu Met Ile Ile His Asp Arg Arg Glu Phe Ala
Lys Phe Glu Lys 755 760 765Glu Lys
Met Asn Ala Lys Trp Asp Thr Gly Glu Asn Pro Ile Tyr Lys 770
775 780Ser Ala Val Thr Thr Val Val Asn Pro Lys Tyr
Glu Gly Lys785 790 795601253PRTHomo
sapiens 60Met Leu Ser Leu Arg Leu Gln Thr Gly Pro Glu Lys Ala Glu Leu
Gln1 5 10 15Glu Leu Asn
Ala Arg Leu Tyr Asp Tyr Val Cys Arg Val Arg Glu Leu 20
25 30Glu Arg Glu Asn Leu Leu Leu Glu Glu Glu
Leu Arg Gly Arg Arg Gly 35 40
45Arg Glu Gly Leu Trp Ala Glu Gly Gln Ala Arg Cys Ala Glu Glu Ala 50
55 60Arg Ser Leu Arg Gln Gln Leu Asp Glu
Leu Ser Trp Ala Thr Ala Leu65 70 75
80Ala Glu Gly Glu Arg Asp Ala Leu Arg Arg Glu Leu Arg Glu
Leu Gln 85 90 95Arg Leu
Asp Ala Glu Glu Arg Ala Ala Arg Gly Arg Leu Asp Ala Glu 100
105 110Leu Gly Ala Gln Gln Arg Glu Leu Gln
Glu Ala Leu Gly Ala Arg Ala 115 120
125Ala Leu Glu Ala Leu Leu Gly Arg Leu Gln Ala Glu Arg Arg Gly Leu
130 135 140Asp Ala Ala His Glu Arg Asp
Val Arg Glu Leu Arg Ala Arg Ala Ala145 150
155 160Ser Leu Thr Met His Phe Arg Ala Arg Ala Thr Gly
Pro Ala Ala Pro 165 170
175Pro Pro Arg Leu Arg Glu Val His Asp Ser Tyr Ala Leu Leu Val Ala
180 185 190Glu Ser Trp Arg Glu Thr
Val Gln Leu Tyr Glu Asp Glu Val Arg Glu 195 200
205Leu Glu Glu Ala Leu Arg Arg Gly Gln Glu Ser Arg Leu Gln
Ala Glu 210 215 220Glu Glu Thr Arg Leu
Cys Ala Gln Glu Ala Glu Ala Leu Arg Arg Glu225 230
235 240Ala Leu Gly Leu Glu Gln Leu Arg Ala Arg
Leu Glu Asp Ala Leu Leu 245 250
255Arg Met Arg Glu Glu Tyr Gly Ile Gln Ala Glu Glu Arg Gln Arg Val
260 265 270Ile Asp Cys Leu Glu
Asp Glu Lys Ala Thr Leu Thr Leu Ala Met Ala 275
280 285Asp Trp Leu Arg Asp Tyr Gln Asp Leu Leu Gln Val
Lys Thr Gly Leu 290 295 300Ser Leu Glu
Val Ala Thr Tyr Arg Ala Leu Leu Glu Gly Glu Ser Asn305
310 315 320Pro Glu Ile Val Ile Trp Ala
Glu His Val Glu Asn Met Pro Ser Glu 325
330 335Phe Arg Asn Lys Ser Tyr His Tyr Thr Asp Ser Leu
Leu Gln Arg Glu 340 345 350Asn
Glu Arg Asn Leu Phe Ser Arg Gln Lys Ala Pro Leu Ala Ser Phe 355
360 365Asn His Ser Ser Ala Leu Tyr Ser Asn
Leu Ser Gly His Arg Gly Ser 370 375
380Gln Thr Gly Thr Ser Ile Gly Gly Asp Ala Arg Arg Gly Phe Leu Gly385
390 395 400Ser Gly Tyr Ser
Ser Ser Ala Thr Thr Gln Gln Glu Asn Ser Tyr Gly 405
410 415Lys Ala Val Ser Ser Gln Thr Asn Val Arg
Thr Phe Ser Pro Thr Tyr 420 425
430Gly Leu Leu Arg Asn Thr Glu Ala Gln Val Lys Thr Phe Pro Asp Arg
435 440 445Pro Lys Ala Gly Asp Thr Arg
Glu Val Pro Val Tyr Ile Gly Glu Asp 450 455
460Ser Thr Ile Ala Arg Glu Ser Tyr Arg Asp Arg Arg Asp Lys Val
Ala465 470 475 480Ala Gly
Ala Ser Glu Ser Thr Arg Ser Asn Glu Arg Thr Val Ile Leu
485 490 495Gly Lys Lys Thr Glu Val Lys
Ala Thr Arg Glu Gln Glu Arg Asn Arg 500 505
510Pro Glu Thr Ile Arg Thr Lys Pro Glu Glu Lys Met Phe Asp
Ser Lys 515 520 525Glu Lys Ala Ser
Glu Glu Arg Asn Leu Arg Trp Glu Glu Leu Thr Lys 530
535 540Leu Asp Lys Glu Ala Arg Gln Arg Glu Ser Gln Gln
Met Lys Glu Lys545 550 555
560Ala Lys Glu Lys Asp Ser Leu Lys Glu Lys Ser Val Arg Glu Arg Glu
565 570 575Val Pro Ile Ser Leu
Glu Val Ser Gln Asp Arg Arg Ala Glu Val Ser 580
585 590Pro Lys Gly Leu Gln Thr Pro Val Lys Asp Ala Gly
Gly Gly Thr Gly 595 600 605Arg Glu
Ala Glu Ala Arg Glu Leu Arg Phe Arg Leu Gly Thr Ser Asp 610
615 620Ala Thr Gly Ser Leu Gln Gly Asp Ser Met Thr
Glu Thr Val Ala Glu625 630 635
640Asn Ile Val Thr Ser Ile Leu Lys Gln Phe Thr Gln Ser Pro Glu Thr
645 650 655Glu Ala Ser Ala
Asp Ser Phe Pro Asp Thr Lys Val Thr Tyr Val Asp 660
665 670Arg Lys Glu Leu Pro Gly Glu Arg Lys Thr Lys
Thr Glu Ile Leu Val 675 680 685Glu
Ser Lys Leu Thr Glu Asp Val Asp Val Ser Asp Glu Ala Gly Leu 690
695 700Asp Tyr Leu Leu Ser Lys Asp Ile Lys Glu
Val Gly Leu Lys Gly Lys705 710 715
720Ser Ala Glu Gln Met Ile Gly Asp Ile Ile Asn Leu Gly Leu Lys
Gly 725 730 735Arg Glu Gly
Arg Ala Lys Val Val Asn Val Glu Ile Val Glu Glu Pro 740
745 750Val Ser Tyr Val Ser Gly Glu Lys Pro Glu
Glu Phe Ser Val Pro Phe 755 760
765Lys Val Glu Glu Val Glu Asp Val Ser Pro Gly Pro Trp Gly Leu Val 770
775 780Lys Glu Glu Glu Gly Tyr Gly Glu
Ser Asp Val Thr Phe Ser Val Asn785 790
795 800Gln His Arg Arg Thr Lys Gln Pro Gln Glu Asn Thr
Thr His Val Glu 805 810
815Glu Val Thr Glu Ala Gly Asp Ser Glu Gly Glu Gln Ser Tyr Phe Val
820 825 830Ser Thr Pro Asp Glu His
Pro Gly Gly His Asp Arg Asp Asp Gly Ser 835 840
845Val Tyr Gly Gln Ile His Ile Glu Glu Glu Ser Thr Ile Arg
Tyr Ser 850 855 860Trp Gln Asp Glu Ile
Val Gln Gly Thr Arg Arg Arg Thr Gln Lys Asp865 870
875 880Gly Ala Val Gly Glu Lys Val Val Lys Pro
Leu Asp Val Pro Ala Pro 885 890
895Ser Leu Glu Gly Asp Leu Gly Ser Thr His Trp Lys Glu Gln Ala Arg
900 905 910Ser Gly Glu Phe His
Ala Glu Pro Thr Val Ile Glu Lys Glu Ile Lys 915
920 925Ile Pro His Glu Phe His Thr Ser Met Lys Gly Ile
Ser Ser Lys Glu 930 935 940Pro Arg Gln
Gln Leu Val Glu Val Ile Gly Gln Leu Glu Glu Thr Leu945
950 955 960Pro Glu Arg Met Arg Glu Glu
Leu Ser Ala Leu Thr Arg Glu Gly Gln 965
970 975Gly Gly Pro Gly Ser Val Ser Val Asp Val Lys Lys
Val Gln Gly Ala 980 985 990Gly
Gly Ser Ser Val Thr Leu Val Ala Glu Val Asn Val Ser Gln Thr 995
1000 1005Val Asp Ala Asp Arg Leu Asp Leu
Glu Glu Val Ser Lys Asp Glu 1010 1015
1020Ala Ser Glu Met Glu Lys Ala Val Glu Ser Val Val Arg Glu Ser
1025 1030 1035Leu Ser Arg Gln Arg Ser
Pro Ala Pro Gly Ser Pro Asp Glu Glu 1040 1045
1050Gly Gly Ala Glu Ala Pro Ala Ala Gly Ile Arg Phe Arg Arg
Trp 1055 1060 1065Ala Thr Arg Glu Leu
Tyr Ile Pro Ser Gly Glu Ser Glu Val Ala 1070 1075
1080Gly Gly Ala Ser His Ser Ser Gly Gln Arg Thr Pro Gln
Gly Pro 1085 1090 1095Val Ser Ala Thr
Val Glu Val Ser Ser Pro Thr Gly Phe Ala Gln 1100
1105 1110Ser Gln Val Leu Glu Asp Val Ser Gln Ala Ala
Arg His Ile Lys 1115 1120 1125Leu Gly
Pro Ser Glu Val Trp Arg Thr Glu Arg Met Ser Tyr Glu 1130
1135 1140Gly Pro Thr Ala Glu Val Val Glu Met Asp
Val Ser Asn Val Glu 1145 1150 1155Ala
Ile Arg Ser Arg Thr Gln Glu Ala Gly Ala Leu Gly Val Ser 1160
1165 1170Asp Arg Gly Ser Trp Arg Asp Ala Asp
Ser Arg Asn Asp Gln Ala 1175 1180
1185Val Gly Val Ser Phe Lys Ala Ser Ala Gly Glu Gly Asp Gln Ala
1190 1195 1200His Arg Glu Gln Gly Lys
Glu Gln Ala Met Phe Asp Lys Lys Val 1205 1210
1215Gln Leu Gln Arg Met Val Asp Gln Arg Ser Val Ile Ser Asp
Glu 1220 1225 1230Lys Lys Val Ala Leu
Leu Tyr Leu Asp Asn Glu Glu Glu Glu Asn 1235 1240
1245Asp Gly His Trp Phe 125061269PRTHomo sapiens 61Met
Ser Ala Ala Leu Phe Ser Leu Asp Gly Pro Ala Arg Gly Ala Pro1
5 10 15Trp Pro Ala Glu Pro Ala Pro
Phe Tyr Glu Pro Gly Arg Ala Gly Lys 20 25
30Pro Gly Arg Gly Ala Glu Pro Gly Ala Leu Gly Glu Pro Gly
Ala Ala 35 40 45Ala Pro Ala Met
Tyr Asp Asp Glu Ser Ala Ile Asp Phe Ser Ala Tyr 50 55
60Ile Asp Ser Met Ala Ala Val Pro Thr Leu Glu Leu Cys
His Asp Glu65 70 75
80Leu Phe Ala Asp Leu Phe Asn Ser Asn His Lys Ala Gly Gly Ala Gly
85 90 95Pro Leu Glu Leu Leu Pro
Gly Gly Pro Ala Arg Pro Leu Gly Pro Gly 100
105 110Pro Ala Ala Pro Arg Leu Leu Lys Arg Glu Pro Asp
Trp Gly Asp Gly 115 120 125Asp Ala
Pro Gly Ser Leu Leu Pro Ala Gln Val Ala Ala Cys Ala Gln 130
135 140Thr Val Val Ser Leu Ala Ala Ala Gly Gln Pro
Thr Pro Pro Thr Ser145 150 155
160Pro Glu Pro Pro Arg Ser Ser Pro Arg Gln Thr Pro Ala Pro Gly Pro
165 170 175Ala Arg Glu Lys
Ser Ala Gly Lys Arg Gly Pro Asp Arg Gly Ser Pro 180
185 190Glu Tyr Arg Gln Arg Arg Glu Arg Asn Asn Ile
Ala Val Arg Lys Ser 195 200 205Arg
Asp Lys Ala Lys Arg Arg Asn Gln Glu Met Gln Gln Lys Leu Val 210
215 220Glu Leu Ser Ala Glu Asn Glu Lys Leu His
Gln Arg Val Glu Gln Leu225 230 235
240Thr Arg Asp Leu Ala Gly Leu Arg Gln Phe Phe Lys Gln Leu Pro
Ser 245 250 255Pro Pro Phe
Leu Pro Ala Ala Gly Thr Ala Asp Cys Arg 260
26562241PRTHomo sapiens 62Met His Ile Thr Gln Leu Asn Arg Glu Cys Leu Leu
His Leu Phe Ser1 5 10
15Phe Leu Asp Lys Asp Ser Arg Lys Ser Leu Ala Arg Thr Cys Ser Gln
20 25 30Leu His Asp Val Phe Glu Asp
Pro Ala Leu Trp Ser Leu Leu His Phe 35 40
45Arg Ser Leu Thr Glu Leu Gln Lys Asp Asn Phe Leu Leu Gly Pro
Ala 50 55 60Leu Arg Ser Leu Ser Ile
Cys Trp His Ser Ser Arg Val Gln Val Cys65 70
75 80Ser Ile Glu Asp Trp Leu Lys Ser Ala Phe Gln
Arg Ser Ile Cys Ser 85 90
95Arg His Glu Ser Leu Val Asn Asp Phe Leu Leu Arg Val Cys Asp Arg
100 105 110Leu Ser Ala Val Arg Ser
Pro Arg Arg Arg Glu Ala Pro Ala Pro Ser 115 120
125Ser Gly Thr Pro Ile Ala Val Gly Pro Lys Ser Pro Arg Trp
Gly Gly 130 135 140Pro Asp His Ser Glu
Phe Ala Asp Leu Arg Ser Gly Val Thr Gly Ala145 150
155 160Arg Ala Ala Ala Arg Arg Gly Leu Gly Ser
Leu Arg Ala Glu Arg Pro 165 170
175Ser Glu Thr Pro Pro Ala Pro Gly Val Ser Trp Gly Pro Pro Pro Pro
180 185 190Gly Ala Pro Val Val
Ile Ser Val Lys Gln Glu Glu Gly Lys Gln Gly 195
200 205Arg Thr Gly Arg Arg Ser His Arg Ala Ala Pro Pro
Cys Gly Phe Ala 210 215 220Arg Thr Arg
Val Cys Pro Pro Thr Phe Pro Gly Ala Asp Ala Phe Pro225
230 235 240Gln63450PRTHomo sapiens 63Met
Arg Glu Ile Val His Ile Gln Ala Gly Gln Cys Gly Asn Gln Ile1
5 10 15Gly Ala Lys Phe Trp Glu Val
Ile Ser Asp Glu His Gly Ile Asp Pro 20 25
30Ser Gly Asn Tyr Val Gly Asp Ser Asp Leu Gln Leu Glu Arg
Ile Ser 35 40 45Val Tyr Tyr Asn
Glu Ala Ser Ser His Lys Tyr Val Pro Arg Ala Ile 50 55
60Leu Val Asp Leu Glu Pro Gly Thr Met Asp Ser Val Arg
Ser Gly Ala65 70 75
80Phe Gly His Leu Phe Arg Pro Asp Asn Phe Ile Phe Gly Gln Ser Gly
85 90 95Ala Gly Asn Asn Trp Ala
Lys Gly His Tyr Thr Glu Gly Ala Glu Leu 100
105 110Val Asp Ser Val Leu Asp Val Val Arg Lys Glu Cys
Glu Asn Cys Asp 115 120 125Cys Leu
Gln Gly Phe Gln Leu Thr His Ser Leu Gly Gly Gly Thr Gly 130
135 140Ser Gly Met Gly Thr Leu Leu Ile Ser Lys Val
Arg Glu Glu Tyr Pro145 150 155
160Asp Arg Ile Met Asn Thr Phe Ser Val Val Pro Ser Pro Lys Val Ser
165 170 175Asp Thr Val Val
Glu Pro Tyr Asn Ala Thr Leu Ser Ile His Gln Leu 180
185 190Val Glu Asn Thr Asp Glu Thr Tyr Cys Ile Asp
Asn Glu Ala Leu Tyr 195 200 205Asp
Ile Cys Phe Arg Thr Leu Lys Leu Ala Thr Pro Thr Tyr Gly Asp 210
215 220Leu Asn His Leu Val Ser Ala Thr Met Ser
Gly Val Thr Thr Ser Leu225 230 235
240Arg Phe Pro Gly Gln Leu Asn Ala Asp Leu Arg Lys Leu Ala Val
Asn 245 250 255Met Val Pro
Phe Pro Arg Leu His Phe Phe Met Pro Gly Phe Ala Pro 260
265 270Leu Thr Ala Arg Gly Ser Gln Gln Tyr Arg
Ala Leu Thr Val Pro Glu 275 280
285Leu Thr Gln Gln Met Phe Asp Ala Lys Asn Met Met Ala Ala Cys Asp 290
295 300Pro Arg His Gly Arg Tyr Leu Thr
Val Ala Thr Val Phe Arg Gly Arg305 310
315 320Met Ser Met Lys Glu Val Asp Glu Gln Met Leu Ala
Ile Gln Ser Lys 325 330
335Asn Ser Ser Tyr Phe Val Glu Trp Ile Pro Asn Asn Val Lys Val Ala
340 345 350Val Cys Asp Ile Pro Pro
Arg Gly Leu Lys Met Ser Ser Thr Phe Ile 355 360
365Gly Asn Ser Thr Ala Ile Gln Glu Leu Phe Lys Arg Ile Ser
Glu Gln 370 375 380Phe Thr Ala Met Phe
Arg Arg Lys Ala Phe Leu His Trp Tyr Thr Gly385 390
395 400Glu Gly Met Asp Glu Met Glu Phe Thr Glu
Ala Glu Ser Asn Met Asn 405 410
415Asp Leu Val Ser Glu Tyr Gln Gln Tyr Gln Asp Ala Thr Ala Glu Glu
420 425 430Glu Gly Glu Met Tyr
Glu Asp Asp Glu Glu Glu Ser Glu Ala Gln Gly 435
440 445Pro Lys 45064361PRTHomo sapiens 64Met Trp Val
Leu Thr Pro Ala Ala Phe Ala Gly Lys Leu Leu Ser Val1 5
10 15Phe Arg Gln Pro Leu Ser Ser Leu Trp
Arg Ser Leu Val Pro Leu Phe 20 25
30Cys Trp Leu Arg Ala Thr Phe Trp Leu Leu Ala Thr Lys Arg Arg Lys
35 40 45Gln Gln Leu Val Leu Arg Gly
Pro Asp Glu Thr Lys Glu Glu Glu Glu 50 55
60Asp Pro Pro Leu Pro Thr Thr Pro Thr Ser Val Asn Tyr His Phe Thr65
70 75 80Arg Gln Cys Asn
Tyr Lys Cys Gly Phe Cys Phe His Thr Ala Lys Thr 85
90 95Ser Phe Val Leu Pro Leu Glu Glu Ala Lys
Arg Gly Leu Leu Leu Leu 100 105
110Lys Glu Ala Gly Met Glu Lys Ile Asn Phe Ser Gly Gly Glu Pro Phe
115 120 125Leu Gln Asp Arg Gly Glu Tyr
Leu Gly Lys Leu Val Arg Phe Cys Lys 130 135
140Val Glu Leu Arg Leu Pro Ser Val Ser Ile Val Ser Asn Gly Ser
Leu145 150 155 160Ile Arg
Glu Arg Trp Phe Gln Asn Tyr Gly Glu Tyr Leu Asp Ile Leu
165 170 175Ala Ile Ser Cys Asp Ser Phe
Asp Glu Glu Val Asn Val Leu Ile Gly 180 185
190Arg Gly Gln Gly Lys Lys Asn His Val Glu Asn Leu Gln Lys
Leu Arg 195 200 205Arg Trp Cys Arg
Asp Tyr Arg Val Ala Phe Lys Ile Asn Ser Val Ile 210
215 220Asn Arg Phe Asn Val Glu Glu Asp Met Thr Glu Gln
Ile Lys Ala Leu225 230 235
240Asn Pro Val Arg Trp Lys Val Phe Gln Cys Leu Leu Ile Glu Gly Glu
245 250 255Asn Cys Gly Glu Asp
Ala Leu Arg Glu Ala Glu Arg Phe Val Ile Gly 260
265 270Asp Glu Glu Phe Glu Arg Phe Leu Glu Arg His Lys
Glu Val Ser Cys 275 280 285Leu Val
Pro Glu Ser Asn Gln Lys Met Lys Asp Ser Tyr Leu Ile Leu 290
295 300Asp Glu Tyr Met Arg Phe Leu Asn Cys Arg Lys
Gly Arg Lys Asp Pro305 310 315
320Ser Lys Ser Ile Leu Asp Val Gly Val Glu Glu Ala Ile Lys Phe Ser
325 330 335Gly Phe Asp Glu
Lys Met Phe Leu Lys Arg Gly Gly Lys Tyr Ile Trp 340
345 350Ser Lys Ala Asp Leu Lys Leu Asp Trp
355 360651589PRTHomo sapiens 65Met Ala Lys Gln Leu Asn
Leu Pro Glu Asn Thr Asp Asp Trp Thr Lys1 5
10 15Glu Asp Val Asn Gln Trp Leu Glu Ser His Lys Ile
Asp Gln Lys His 20 25 30Arg
Glu Ile Leu Thr Glu Gln Asp Val Asn Gly Ala Val Leu Lys Trp 35
40 45Leu Lys Lys Glu His Leu Val Asp Met
Gly Ile Thr His Gly Pro Ala 50 55
60Ile Gln Ile Glu Glu Leu Phe Lys Glu Leu Arg Lys Thr Ala Ile Glu65
70 75 80Asp Ser Ile Gln Thr
Ser Lys Met Gly Lys Pro Ser Lys Asn Ala Pro 85
90 95Lys Asp Gln Thr Val Ser Gln Lys Glu Arg Arg
Glu Thr Ser Lys Gln 100 105
110Lys Gln Lys Gly Lys Glu Asn Pro Asp Met Ala Asn Pro Ser Ala Met
115 120 125Ser Thr Thr Ala Lys Gly Ser
Lys Ser Leu Lys Val Glu Leu Ile Glu 130 135
140Asp Lys Ile Asp Tyr Thr Lys Glu Arg Gln Pro Ser Ile Asp Leu
Thr145 150 155 160Cys Val
Ser Tyr Pro Phe Asp Glu Phe Ser Asn Pro Tyr Arg Tyr Lys
165 170 175Leu Asp Phe Ser Leu Gln Pro
Glu Thr Gly Pro Gly Asn Leu Ile Asp 180 185
190Pro Ile His Glu Phe Lys Ala Phe Thr Asn Thr Ala Thr Ala
Thr Glu 195 200 205Glu Asp Val Lys
Met Lys Phe Ser Asn Glu Val Phe Arg Phe Ala Ser 210
215 220Ala Cys Met Asn Ser Arg Thr Asn Gly Thr Ile His
Phe Gly Val Lys225 230 235
240Asp Lys Pro His Gly Lys Ile Val Gly Ile Lys Val Thr Asn Asp Thr
245 250 255Lys Glu Ala Leu Ile
Asn His Phe Asn Leu Met Ile Asn Lys Tyr Phe 260
265 270Glu Asp His Gln Val Gln Gln Ala Lys Lys Cys Ile
Arg Glu Pro Arg 275 280 285Phe Val
Glu Val Leu Leu Pro Asn Ser Thr Leu Ser Asp Arg Phe Val 290
295 300Ile Glu Val Asp Ile Ile Pro Gln Phe Ser Glu
Cys Gln Tyr Asp Tyr305 310 315
320Phe Gln Ile Lys Met Gln Asn Tyr Asn Asn Lys Ile Trp Glu Gln Ser
325 330 335Lys Lys Phe Ser
Leu Phe Val Arg Asp Gly Thr Ser Ser Lys Asp Ile 340
345 350Thr Lys Asn Lys Val Asp Phe Arg Ala Phe Lys
Ala Asp Phe Lys Thr 355 360 365Leu
Ala Glu Ser Arg Lys Ala Ala Glu Glu Lys Phe Arg Ala Lys Thr 370
375 380Asn Lys Lys Glu Arg Glu Gly Pro Lys Leu
Val Lys Leu Leu Thr Gly385 390 395
400Asn Gln Asp Leu Leu Asp Asn Ser Tyr Tyr Glu Gln Tyr Ile Leu
Val 405 410 415Thr Asn Lys
Cys His Pro Asp Gln Thr Lys His Leu Asp Phe Leu Lys 420
425 430Glu Ile Lys Trp Phe Ala Val Leu Glu Phe
Asp Pro Glu Ser Asn Ile 435 440
445Asn Gly Val Val Lys Ala Tyr Lys Glu Ser Arg Val Ala Asn Leu His 450
455 460Phe Pro Ser Val Tyr Val Glu Gln
Lys Thr Thr Pro Asn Glu Thr Ile465 470
475 480Ser Thr Leu Asn Leu Tyr His Gln Pro Ser Trp Ile
Phe Cys Asn Gly 485 490
495Arg Leu Asp Leu Asp Ser Glu Lys Tyr Lys Pro Phe Asp Pro Ser Ser
500 505 510Trp Gln Arg Glu Arg Ala
Ser Asp Val Arg Lys Leu Ile Ser Phe Leu 515 520
525Thr His Glu Asp Ile Met Pro Arg Gly Lys Phe Leu Val Val
Phe Leu 530 535 540Leu Leu Ser Ser Val
Asp Asp Pro Arg Asp Pro Leu Ile Glu Thr Phe545 550
555 560Cys Ala Phe Tyr Gln Asp Leu Lys Gly Met
Glu Asn Ile Leu Cys Ile 565 570
575Cys Val His Pro His Ile Phe Gln Gly Trp Lys Asp Leu Leu Glu Ala
580 585 590Arg Leu Ile Lys His
Gln Asp Glu Ile Ser Ser Gln Cys Ile Ser Ala 595
600 605Leu Ser Leu Glu Glu Ile Asn Gly Thr Ile Leu Lys
Leu Lys Ser Val 610 615 620Thr Gln Ser
Ser Lys Arg Leu Leu Pro Ser Ile Gly Leu Ser Thr Val625
630 635 640Leu Leu Lys Lys Glu Glu Asp
Ile Met Thr Ala Leu Glu Ile Ile Cys 645
650 655Glu Asn Glu Cys Glu Gly Thr Leu Leu Glu Lys Asp
Lys Asn Lys Phe 660 665 670Leu
Glu Phe Lys Ala Ser Lys Glu Glu Asp Phe Tyr Arg Gly Gly Lys 675
680 685Val Ser Trp Trp Asn Phe Tyr Phe Ser
Ser Glu Ser Tyr Ser Ser Pro 690 695
700Phe Val Lys Arg Asp Lys Tyr Glu Arg Leu Glu Ala Met Ile Gln Asn705
710 715 720Cys Ala Asp Ser
Ser Lys Pro Thr Ser Thr Lys Ile Ile His Leu Tyr 725
730 735His His Pro Gly Cys Gly Gly Thr Thr Leu
Ala Met His Ile Leu Trp 740 745
750Glu Leu Arg Lys Lys Phe Arg Cys Ala Val Leu Lys Asn Lys Thr Val
755 760 765Asp Phe Ser Glu Ile Gly Glu
Gln Val Thr Ser Leu Ile Thr Tyr Gly 770 775
780Ala Met Asn Arg Gln Glu Tyr Val Pro Val Leu Leu Leu Val Asp
Asp785 790 795 800Phe Glu
Glu Gln Asp Asn Val Tyr Leu Leu Gln Tyr Ser Ile Gln Thr
805 810 815Ala Ile Ala Lys Lys Tyr Ile
Arg Tyr Glu Lys Pro Leu Val Ile Ile 820 825
830Leu Asn Cys Met Arg Ser Gln Asn Pro Glu Lys Ser Ala Arg
Ile Pro 835 840 845Asp Ser Ile Ala
Val Ile Gln Gln Leu Ser Pro Lys Glu Gln Arg Ala 850
855 860Phe Glu Leu Lys Leu Lys Glu Ile Lys Glu Gln His
Lys Asn Phe Glu865 870 875
880Asp Phe Tyr Ser Phe Met Ile Met Lys Thr Asn Phe Asn Lys Glu Tyr
885 890 895Ile Glu Asn Val Val
Arg Asn Ile Leu Lys Gly Gln Asn Ile Phe Thr 900
905 910Lys Glu Ala Lys Leu Phe Ser Phe Leu Ala Leu Leu
Asn Ser Tyr Val 915 920 925Pro Asp
Thr Thr Ile Ser Leu Ser Gln Cys Glu Lys Phe Leu Gly Ile 930
935 940Gly Asn Lys Lys Ala Phe Trp Gly Thr Glu Lys
Phe Glu Asp Lys Met945 950 955
960Gly Thr Tyr Ser Thr Ile Leu Ile Lys Thr Glu Val Ile Glu Cys Gly
965 970 975Asn Tyr Cys Gly
Val Arg Ile Ile His Ser Leu Ile Ala Glu Phe Ser 980
985 990Leu Glu Glu Leu Lys Lys Ser Tyr His Leu Asn
Lys Ser Gln Ile Met 995 1000
1005Leu Asp Met Leu Thr Glu Asn Leu Phe Phe Asp Thr Gly Met Gly
1010 1015 1020Lys Ser Lys Phe Leu Gln
Asp Met His Thr Leu Leu Leu Thr Arg 1025 1030
1035His Arg Asp Glu His Glu Gly Glu Thr Gly Asn Trp Phe Ser
Pro 1040 1045 1050Phe Ile Glu Ala Leu
His Lys Asp Glu Gly Asn Glu Ala Val Glu 1055 1060
1065Ala Val Leu Leu Glu Ser Ile His Arg Phe Asn Pro Asn
Ala Phe 1070 1075 1080Ile Cys Gln Ala
Leu Ala Arg His Phe Tyr Ile Lys Lys Lys Asp 1085
1090 1095Phe Gly Asn Ala Leu Asn Trp Ala Lys Gln Ala
Lys Ile Ile Glu 1100 1105 1110Pro Asp
Asn Ser Tyr Ile Ser Asp Thr Leu Gly Gln Val Tyr Lys 1115
1120 1125Ser Lys Ile Arg Trp Trp Ile Glu Glu Asn
Gly Gly Asn Gly Asn 1130 1135 1140Ile
Ser Val Asp Asp Leu Ile Ala Leu Leu Asp Leu Ala Glu His 1145
1150 1155Ala Ser Ser Ala Phe Lys Glu Ser Gln
Gln Gln Ser Glu Asp Arg 1160 1165
1170Glu Tyr Glu Val Lys Glu Arg Leu Tyr Pro Lys Ser Lys Arg Arg
1175 1180 1185Tyr Asp Thr Tyr Asn Ile
Ala Gly Tyr Gln Gly Glu Ile Glu Val 1190 1195
1200Gly Leu Tyr Thr Ile Gln Ile Leu Gln Leu Ile Pro Phe Phe
Asp 1205 1210 1215Asn Lys Asn Glu Leu
Ser Lys Arg Tyr Met Val Asn Phe Val Ser 1220 1225
1230Gly Ser Ser Asp Ile Pro Gly Asp Pro Asn Asn Glu Tyr
Lys Leu 1235 1240 1245Ala Leu Lys Asn
Tyr Ile Pro Tyr Leu Thr Lys Leu Lys Phe Ser 1250
1255 1260Leu Lys Lys Ser Phe Asp Phe Phe Asp Glu Tyr
Phe Val Leu Leu 1265 1270 1275Lys Pro
Arg Asn Asn Ile Lys Gln Asn Glu Glu Ala Lys Thr Arg 1280
1285 1290Arg Lys Val Ala Gly Tyr Phe Lys Lys Tyr
Val Asp Ile Phe Cys 1295 1300 1305Leu
Leu Glu Glu Ser Gln Asn Asn Thr Gly Leu Gly Ser Lys Phe 1310
1315 1320Ser Glu Pro Leu Gln Val Glu Arg Cys
Arg Arg Asn Leu Val Ala 1325 1330
1335Leu Lys Ala Asp Lys Phe Ser Gly Leu Leu Glu Tyr Leu Ile Lys
1340 1345 1350Ser Gln Glu Asp Ala Ile
Ser Thr Met Lys Cys Ile Val Asn Glu 1355 1360
1365Tyr Thr Phe Leu Leu Glu Gln Cys Thr Val Lys Ile Gln Ser
Lys 1370 1375 1380Glu Lys Leu Asn Phe
Ile Leu Ala Asn Ile Ile Leu Ser Cys Ile 1385 1390
1395Gln Pro Thr Ser Arg Leu Val Lys Pro Val Glu Lys Leu
Lys Asp 1400 1405 1410Gln Leu Arg Glu
Val Leu Gln Pro Ile Gly Leu Thr Tyr Gln Phe 1415
1420 1425Ser Glu Pro Tyr Phe Leu Ala Ser Leu Leu Phe
Trp Pro Glu Asn 1430 1435 1440Gln Gln
Leu Asp Gln His Ser Glu Gln Met Lys Glu Tyr Ala Gln 1445
1450 1455Ala Leu Lys Asn Ser Phe Lys Gly Gln Tyr
Lys His Met His Arg 1460 1465 1470Thr
Lys Gln Pro Ile Ala Tyr Phe Phe Leu Gly Lys Gly Lys Arg 1475
1480 1485Leu Glu Arg Leu Val His Lys Gly Lys
Ile Asp Gln Cys Phe Lys 1490 1495
1500Lys Thr Pro Asp Ile Asn Ser Leu Trp Gln Ser Gly Asp Val Trp
1505 1510 1515Lys Glu Glu Lys Val Gln
Glu Leu Leu Leu Arg Leu Gln Gly Arg 1520 1525
1530Ala Glu Asn Asn Cys Leu Tyr Ile Glu Tyr Gly Ile Asn Glu
Lys 1535 1540 1545Ile Thr Ile Pro Ile
Thr Pro Ala Phe Leu Gly Gln Leu Arg Ser 1550 1555
1560Gly Arg Ser Ile Glu Lys Val Ser Phe Tyr Leu Gly Phe
Ser Ile 1565 1570 1575Gly Gly Pro Leu
Ala Tyr Asp Ile Glu Ile Val 1580 158566122PRTHomo
sapiens 66Met Glu Ala Ser Ala Leu Thr Ser Ser Ala Val Thr Ser Val Ala
Lys1 5 10 15Val Val Arg
Val Ala Ser Gly Ser Ala Val Val Leu Pro Leu Ala Arg 20
25 30Ile Ala Thr Val Val Ile Gly Gly Val Val
Ala Met Ala Ala Val Pro 35 40
45Met Val Leu Ser Ala Met Gly Phe Thr Ala Ala Gly Ile Ala Ser Ser 50
55 60Ser Ile Ala Ala Lys Met Met Ser Ala
Ala Ala Ile Ala Asn Gly Gly65 70 75
80Gly Val Ala Ser Gly Ser Leu Val Ala Thr Leu Gln Ser Leu
Gly Ala 85 90 95Thr Gly
Leu Ser Gly Leu Thr Lys Phe Ile Leu Gly Ser Ile Gly Ser 100
105 110Ala Ile Ala Ala Val Ile Ala Arg Phe
Tyr 115 120
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