Patent application title: UNIQUE ASSOCIATED KAPOSI'S SARCOMA VIRUS SEQUENCES AND USES THEREOF
Yuan Chang (New York, NY, US)
Patrick S. Moore (New York, NY, US)
IPC8 Class: AA01K67033FI
Class name: Multicellular living organisms and unmodified parts thereof and related processes nonhuman animal transgenic nonhuman animal (e.g., mollusks, etc.)
Publication date: 2011-10-27
Patent application number: 20110265195
This invention provides an isolated peptide encoded by a nucleic acid
which is at least 30 nucleotides in length and has a sequence which
uniquely defines a herpesvirus associated with Kaposis' sarcoma, which
herpesvirus is present in and recoverable from the HBL-6 cell line (ATCC
Accession No. CRL 11762).
43. An antisense molecule capable of hybridizing to an isolated nucleic acid which uniquely defines a herpesvirus associated with Kaposi's sarcoma and which comprises at least 30 nucleotides of a nucleic acid selected from the group consisting of SEQ ID Nos: 1-15.
44. The antisense molecule of claim 43, wherein the molecule is a DNA.
45. The antisense molecule of claim 43, wherein the molecule is a RNA.
46. A triplex oligonucleotide capable of hybridizing with a double stranded isolated nucleic acid which uniquely defines a herpesvirus associated with Kaposi's sarcoma and which comprises at least 30 nucleotides of a nucleic acid selected from the group consisting of SEQ ID Nos: 1-15.
47. A transgenic nonhuman mammal which comprises at least a portion of an isolated nucleic acid which uniquely defines a herpesvirus associated with Kaposi's sarcoma and which comprises at least 30 nucleotides of a nucleic acid selected from the group consisting of SEQ ID Nos: 1-15, introduced into the mammal at an embryonic stage.
48. An antisense molecule capable of hybridizing to an isolated nucleic acid which is at least 30 nucleotides in length and has a sequence which uniquely defines a herpesvirus associated with Kaposi's sarcoma, which herpesvirus is present in and recoverable from the HBL-6 cell line (ATCC Accession No. CRL 11762).
49. The antisense molecule of claim 48, wherein the molecule is a DNA.
50. The antisense molecule of claim 48, wherein the molecule is a RNA.
51. A triplex oligonucleotide capable of hybridizing with a double isolated nucleic acid which is at least 30 nucleotides in length and has a sequence which uniquely defines a herpesvirus associated with Kaposi's sarcoma, which herpesvirus is present in and recoverable from the HBL-6 cell line (ATCC Accession No. CRL 11762).
52. A transgenic nonhuman mammal which comprises at least a portion of an isolated nucleic acid which is at least 30 nucleotides in length and has a sequence which uniquely defines a herpesvirus associated with Kaposi's sarcoma, which herpesvirus is present in and recoverable from the HBL-6 cell line (ATCC Accession No. CRL 11762) introduced into the mammal at an embryonic stage.
53. A method of prophylaxis or treatment for Kaposi's sarcoma (KS) by administering to a subject at risk for KS, an antibody that binds to an isolated DNA virus associated with Kaposi's Sarcoma wherein the viral DNA: (a) encodes a thymidine kinase; and (b) hybridizes under conditions of high stringency with a nucleic acid selected from the group consisting of SEQ ID Nos: 1-15 in a pharmaceutically acceptable carrier.
54. A method of immunizing a subject against a disease caused by the herpesvirus associated with Kaposi's sarcoma which comprises administering to the subject an effective immunizing dose of the vaccine of an associated herpesvirus associated with Kaposi's sarcoma comprising an isolated nucleic acid which uniquely defines a herpesvirus associated with Kaposi's sarcoma and which comprises at least 30 nucleotides of a nucleic acid selected from the group consisting of SEQ ID Nos: 1-15.
55. A method for preventing the development or transmission of herpesvirus associated Kaposi's sarcoma in a subject by treating a subject with Kaposi's sarcoma (KS) comprising administering to the subject having a human herpesvirus-associated KS a pharmaceutically effective amount of an antiviral agent in a pharmaceutically acceptable carrier, wherein the agent is effective to prevent the development or transmission of an isolated DNA virus associated with Kaposi's Sarcoma wherein the viral DNA: (a) encodes a thymidine kinase; and (b) hybridizes under conditions of high stringency with a nucleic acid selected from the group consisting of SEQ ID Nos: 1-15.
56. An isolated Kaposi's sarcoma-associated herpesvirus (KSHV) glycoprotein H (gH) polypeptide.
57. The isolated polypeptide of claim 56, wherein the polypeptide is linked to a second polypeptide to form a fusion protein.
58. The fusion protein of claim 57, wherein the second polypeptide is beta-galactosidase.
59. An antibody which specifically binds to the polypeptide of claim 56.
60. The antibody of claim 59, wherein the antibody is polyclonal antibody.
61. The antibody of claim 59, wherein the antibody is a monoclonal antibody.
62. An antisense molecule capable of hybridizing to an isolated nucleic acid molecule encoding Kaposi's sarcoma-associated herpesvirus (KSHV) glycoprotein H (gH).
63. The antisense molecule of claim 62, wherein the molecule is a nucleic acid derivative.
64. The antisense molecule of claim 62, wherein the molecule is an RNA derivative.
65. A triplex oligonucleotide capable of hybridizing with an isolated nucleic acid molecule encoding Kaposi's sarcoma-associated herpesvirus (KSHV) glycoprotein H (gH).
66. A peptide conjugated to a carrier protein wherein the peptide is encoded by at least 30 nucleotides and has a sequence which is encoded by, and uniquely defines, Kaposi's sarcoma herpesvirus (KSHV), (deposited in BHL-6 cells under ATCC Accession No. 11762), wherein the sequence of the peptide is present within the amino acid sequence of any of SEQ ID NOS: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33 or 35.
 This application is a divisional of U.S. Ser. No. 11/801,641, filed
May 9, 2007, which is a continuation of U.S. Ser. No. 09/607,179, filed
Jun. 29, 2000, which is a continuation of U.S. Ser. No. 08/793,624, filed
Feb. 18, 1997, now U.S. Pat. No. 6,150,093, issued Nov. 21, 2000, which
is a 5371 national stage application of PCT International Application No.
PCT/US95/10194, filed Aug. 11, 1995, which is a continuation-in-part of
U.S. Ser. No. 08/420,235, filed Apr. 11, 1995, now U.S. Pat. No.
5,801,042, issued Sep. 1, 1998, which is a continuation-in-part of U.S.
Ser. No. 08/343,101, filed Nov. 21, 1994, now U.S. Pat. No. 5,830,759,
issued Nov. 3, 1998, which is a continuation-in-part of U.S. Ser. No.
08/292,365, filed Aug. 18, 1994, now abandoned, the contents all of which
are hereby incorporated by reference in their entireties into this
 This application incorporates-by-reference nucleotide and/or amino acid sequences which are present in the file named "110426--0575--45185-CAAZ-PCT-US_SubSequenceListingAHC.txt" which is 122 kilobytes in size, and which was created Apr. 26, 2011, in the IBM-PCT machine format, having an operating system compatibility with MS-Windows, which is contained in the text file filed Apr. 26, 2011 as a part of this application.
 Throughout this application, various publications may be referenced by Arabic numerals in brackets. Full citations for these publications may be found at the end of each Experimental Details Section. The disclosures of the publications cited herein are in their entirety hereby incorporated by reference into this application to more fully describe the state of the art to which this invention pertains.
BACKGROUND OF THE INVENTION
 Kaposi's sarcoma (KS) is the most common neoplasm occurring in persons with acquired immunodeficiency syndrome (AIDS). Approximately 15-20% of AIDS patients develop this neoplasm which rarely occurs in immunocompetent individuals [13, 14]. Epidemiologic evidence suggests that AIDS-associated KS (AIDS-KS) has an infectious etiology. Gay and bisexual AIDS patients are approximately twenty times more likely than hemophiliac AIDS patients to develop KS, and KS may be associated with specific sexual practices among gay men with AIDS [6, 15, 55, 83]. KS is uncommon among adult AIDS patients infected through heterosexual or parenteral HIV transmission, or among pediatric AIDS patients infected through vertical HIV transmission . Agents previously suspected of causing KS include cytomegalovirus, hepatitis B virus, human papillomavirus, Epstein-Barr virus, human herpesvirus 6, human immunodeficiency virus (HIV), and Mycoplasma penetrans [18, 23, 85, 91, 92]. Non-infectious environmental agents, such as nitrite inhalants, also have been proposed to play a role in KS tumorigenesis . Extensive investigations, however, have not demonstrated an etiologic association between any of these agents and AIDS-KS [37, 44, 46, 90].
SUMMARY OF THE INVENTION
 This invention provides an isolated DNA molecule which is at least 30 nucleotides in length and which uniquely defines a herpesvirus associated with Kaposi's sarcoma. This invention provides an isolated herpesvirus associated with Kaposi's sarcoma.
 This invention provides a method of vaccinating a subject for KS, prophylaxis diagnosing or treating a subject with KS and detecting expression of a DNA virus associated with Kaposi's sarcoma in a cell.
BRIEF DESCRIPTION OF THE FIGURES
 FIG. 1:
 Agarose gel electrophoresis of RDA products from AIDS-KS tissue and uninvolved tissue. RDA was performed on DNA extracted from KS skin tissue and uninvolved normal skin tissue obtained at autopsy from a homosexual man with AIDS-KS. Lane 1 shows the initial PCR amplified genomic representation of the AIDS-KS DNA after Bam HI digestion. Lanes 2-4 show that subsequent cycles of ligation, amplification, hybridization and digestion of the RDA products resulted in amplification of discrete bands at 380, 450, 540 and 680 bp. RDA of the extracted AIDS-KS DNA performed against itself resulted in a single band at 540 bp (lane 5). Bands at 380 bp and 680 bp correspond to KS330Bam and KS627Bam respectively after removal of 28 bp priming sequences. Bands at 450 and 540 bp hybridized nonspecifically to both KS and non-KS human DNA. Lane M is a molecular weight marker.
 FIGS. 2A-2B:
 Hybridization of 32P-labelled KS330Bam (FIG. 2A) and KS627Bam (FIG. 2B) sequences to a representative panel of 19 DNA samples extracted from KS lesions and digested with Bam HI. KS330Bam hybridized to 11 of the 19 and KS627Bam hybridized to 12 of the 19 DNA samples from AIDS-KS lesions. Two additional cases (lanes 12 and 13) were shown to have faint bands with both KS330Bam and KS627Bam probes after longer exposure. One negative specimen (lane 3) did not have microscopically detectable KS in the tissue specimen. Seven of 8 additional KS DNA samples also hybridized to both sequences.
 FIGS. 3A-3F:
 Nucleotide sequences of the DNA herpesvirus associated with KS (KSHV). (SEQ ID NOs: 1, 36-40, respectively).
 FIGS. 4A-4B:
 PCR amplification of a representative set of KS-derived DNA samples using KS330234 primers. FIG. 4A shows the agarose gel of the amplification products from 19 KS DNA samples (lanes 1-19) and FIG. 4B, shows specific hybridization of the PCR products to a 32P end-labelled 25 bp internal oligonucleotide (FIG. 3B) after transfer of the gel to a nitrocellulose filter. Negative samples in lanes 3 and 15 respectively lacked microscopically detectable KS in the sample or did not amplify the constitutive p53 exon 6, suggesting that these samples were negative for technical reasons. An additional 8 AIDS-KS samples were amplified and all were positive for KS330234. Lane 20 is a negative control and Lane M is a molecular weight marker.
 FIG. 5:
 Southern blot hybridization of KS330Bam and KS627Bam to AIDS-KS genomic DNA extracted from three subjects (lanes 1, 2, and 3) and digested with PvuII. Based on sequence information (FIG. 3A), restricted sites for Pvu II occur between by 12361-12362 of the KSHV sequence (FIG. 3A, SEQ ID NO: 1), at bp 134 in KS330Bam (FIG. 3B, SEQ ID NO: 36) and bp 414 in KS627Bam (FIG. 3C, SEQ ID NO: 37). KS330Bam and KS627Bam failed to hybridize to the same fragments in the digests indicating that the two sequences are separated from each other by one or more intervening Bam HI restriction fragments. Digestion with Pvu II and hybridization to KS330Bam resulted in two distinct banding patterns (lanes 1 and 2 vs. lane 3) suggesting variation between KS samples.
 FIG. 6:
 Comparison of amino acid homologies between EBV ORF BDLF1 (SEQ ID NO:47), HSVSA ORF 26 (SEQ ID NO:46) and a 918 bp reading frame of the Kaposi's sarcoma agent which includes KS330Bam (SEQ ID NO:25). Amino acid identity is denoted by reverse lettering. In HSVSA, ORF 26 encodes a minor capsid VP23 which is a late gene product.
 FIG. 7:
 Subculture of Raji cells co-cultivated with BCBL-1 cells treated with TPA for 2 days. PCR shows that Raji cells are positive for KSHV sequences and indicate that the agent is a transmissible virus.
 FIG. 8:
 A schematic diagram of the orientation of KSHV open reading frames identified on the KS5 20,710 bp DNA fragment. Homologs to each open reading frame from a corresponding region of the herpesvirus saimiri (HSVSA) genome are present in an identical orientation, except for the region corresponding to the ORF 28 of HSVSA (middle schematic section). The shading for each open reading frame corresponds to the approximate % amino acid identity for the KSHV ORF compared to this homolog in HSVSA. Noteworthy homologs that are present in this section of DNA include homologs to thymidine kinase (ORF21), gH glycoprotein (ORF22), major capsid protein (ORF25) and the VP23 protein (ORF26) which contains the original KS330Bam sequence derived by representational difference analysis.
 FIG. 9:
 The ˜200 kD antigen band appearing on a Western blot of KS patient sera against BCBL1 lysate (B1) and Raji lysate (RA). M is molecular weight marker. The antigen is a doublet between ca. 210 kD and 240 kD.
 FIG. 10:
 5 control patient sera without KS (A1N, A2N, A3N, A4N and A5N). B1=BCBL1 lysate, RA=Raji lysate. The 220 kD band is absent from the Western blots using patient sera without KS.
 FIG. 11:
 In this figure, 0.5 ml aliquots of the gradient have been fractionated (fractions 1-62) with the 30% gradient fraction being at fraction No. 1 and the 10% gradient fraction being at fraction No. 62. Each fraction has been dot hybridized to a nitrocellulose membrane and then a 32P-labeled KSHV DNA fragment, KS631Bam has been hybridized to the membrane using standard techniques. The figure shows that the major solubilized fraction of the KSHV genome bands (i.e. is isolated) in fractions 42 through 48 of the gradient with a high concentration of the genome being present in fraction 44. A second band of solubilized KSHV DNA occurs in fractions 26 through 32.
 FIG. 12:
 Location, feature, and relative homologies of KS5 open reading frames compared to translation products of herpesvirus saimiri (HSV), equine herpesvirus 2 (EHV2) and Epstein-Barr virus (EDV).
 FIG. 13:
 Indirect immunofluorescence end-point and geometric mean titers (GMT) in AIDS-KS and AIDS control sera against HBL-6 and P3H3 prior to and after adsorption with P3H3.
 FIG. 14:
 Genetic map of KS5, a 20.7 kb lambda phage clone insert derived from a human genomic library prepared from an AIDS-KS lesion. Seventeen partial and complete open reading frames (ORFs) are identified with arrows denoting reading frame orientations. Comparable regions of the Epstein-Barr virus (EBV) and herpesvirus saimiri (HVS) genomes are shown for comparison. Levels of amino acid similarity between KSHV ORFs are indicated by shading of EBV and HVS ORFs (black, over 70% similarity; dark gray, 55-70% similarity; light gray, 40-54% similarity; white, no detectable homology). Domains of conserved herpesvirus sequence blocks and locations of restriction endonuclease sites used in subcloning are shown beneath the KSHV map (B, Bam HI site; N, Not I site). The small Bam HI fragment (black) in the VP23 gene homolog corresponds to the KS330Bam fragment generated by representational difference analysis which was used to identify the KS5 lambda phage clone.
 FIGS. 15A-15B:
 Phylogenetic trees of KSHV based on comparison of aligned amino acid sequences between herpesviruses for the MCP gene and for a concatenated nine-gene set. The comparison of MCP sequences (FIG. 15A) was obtained by the neighbor-joining method and is shown in unrooted form with branch lengths proportional to divergence (mean number of substitution events per site) between the nodes bounding each branch. Comparable results were obtained by maximum parsimony analysis. The number of times out of 100 bootstrap samplings the division indicated by each internal branch was obtained are shown next to each branch; bootstrap values below 75 are not shown. FIG. 15B is a phylogenetic tree of gammaherpesvirus sequences based on a nine-gene set CS1 (see text) and demonstrates that KSHV is most closely related to the gamma-2 herpesvirus sublineage, genus Rhadinovirus. The CS1 amino acid sequence was used to infer a tree by the Protml maximum likelihood method; comparable results, not shown were obtained with the neighbor-joining and maximum parsimony methods. The bootstrap value for the central branch is marked. On the basis of the MCP analysis, the root must lie between EBV and the other three species. Abbreviations for virus species used in the sequence comparisons are 1) Alphaherpesvirinae: HSV1 and HSV2, herpes simplex virus types 1 and 2; EHV1, equine herpesvirus 1; PRV, pseudorabies virus; and VZV, varicella-zoster virus, 2) Betaherpesvirinae: HCMV, human cytomegalovirus; HHV6 and HHV7, human herpesviruses 6 and 7, and 3) Gammaherpesvirinae: HVS, herpesvirus saimiri; EHV2, equine herpesvirus 2; EBV, Epstein-Barr virus; and Kaposi's sarcoma-associated herpesvirus.
 FIGS. 16A-16B:
 CHEF gel electrophoresis of BCBL-1 DNA hybridized to KS631Bam (FIG. 16A) and EBV terminal repeat (FIG. 16B). KS631Bam hybridizes to a band at 270 kb as well as to a diffuse band at the origin. The EBV termini sequence hybridizes to a 150-160 kb band consistent with the linear form of the genome. Both KS631Bam (dark arrow) and an EBV terminal sequence hybridize to high molecular weight bands immediately below the origin indicating possible concatemeric or circular DNA. The high molecular weight KS631Bam hybridizing band reproduces poorly but is visible on the original autoradiographs.
 FIG. 17:
 Induction of KSHV and EBV replication in BCBL-1 with increasing concentrations of TPA. Each determination was made in triplicate after 48 h of TPA incubation and hybridization was standardized to the amount of cellular DNA by hybridization to beta-actin. The figure shows the mean and range of relative increase in hybridizing genome for EBV and KSHV induced by TPA compared to uninduced BCBL-1. TPA at 20 ng/ml induced an eight-fold increase in EBV genome (upper line) at 48 h compared to only a 1.4 fold increase in KSHV genome (lower line). Despite the lower level of KSHV induction, increased replication of KSHV genome after induction with TPA concentrations over 10 ng/ml was reproducibly detected.
 FIGS. 18A-18C:
 In situ hybridization with an ORF26 oligomer to BCBL-1, Raji and RCC-1 cells. Hybridization occurred to nuclei of KSHV infected BCBL-1 (FIG. 18A), but not to uninfected Raji cells (FIG. 18B). RCC-1, a Raji cell line derived by cultivation of Raji with BCBL-1 in communicating chambers separated by a 0.4 5μ filter, shows rare cells with positive hybridization to the KSHV ORF26 probe (FIG. 18C).
 FIGS. 19A-19D:
 Representative example of IFA staining of HBL-6 with AIDS-KS patient sera and control sera from HIV-infected patients without KS. Both AIDS-KS (FIG. 19A) and control (FIG. 19B) sera show homogeneous staining of HBL-6 at 1:50 dilution. After adsorption with paraformaldehyde-fixed P3H3 to remove cross-reacting antibodies directed against lymphocyte and EBV antigens, antibodies from AIDS-KS sera localize to HBL-6 nuclei (FIG. 19C). P3H3 adsorption of control sera eliminates immunofluorescent staining of HBL-6).
 FIGS. 20A-20B:
 Longitudinal PCR examination for KSHV DNA of paired PBMC samples from AIDS-KS patients (A) and homosexual/bisexual AIDS patients without KS (B). Time 0 is the date of KS onset for cases or other AIDS-defining illness for controls. All samples were randomized and examined blindly. Overall, 7 of the KS patients were KSHV positive at both examination dates (solid bars) and 5 converted from a negative to positive PBMC sample (forward striped bars) immediately prior to or after KS onset. Two previously positive KS patients were negative after KS diagnosis (reverse striped bars) and the remaining KS patients were negative at both timepoints (open bars). Two KS converted from negative to positive and one control patient reverted from PCR positive to negative for KSHV DNA.
 FIG. 21:
 Sample collection characteristics for AIDS-KS patients, gay/bisexual AIDS patients and hemophilic AIDS patients.
 FIG. 22:
 PCR analysis of KS330233 in DNA samples from patients with Kaposi's sarcoma and tumor controls.
 FIG. 23:
 Characteristics of the study population of patients with KS and without KS.
 FIG. 24:
 Prevalence of antibody to KSHV p40 in HIV-1 positive patients with and without KS.
 FIG. 25:
 Comparison of KS patients with and without antibody to KSHV p40.
 FIG. 26:
 Prevalence of antibody detectable by indirect immunofluorescence to KSHV antigens in chemically induced BCBL-1 cells in HIV-1 positive patients with and without KS.
 FIGS. 27A-27B:
 Specific recognition of KSHV polypeptides in chemically treated BCBL-1 cells. FIG. 27A shows reactivity of untreated BCBL-1 and B95-8 cells with RM, a reference human antibody to EBV. RM recognizes the EBV polypeptides EBNA1 and p21 in the BCBL-1 cells. FIG. 27B shows reactivity of untreated and chemically treated cells with serum 01-03 from a patient with KS. Cells were treated with TPA and n-butyrate for 48 hrs. For description of the cell lines see Materials and Methods. The immunoblots were prepared from 10% SDS polyacrylamide gels.
 FIGS. 28A-28D:
 Detection of KSHV p40 by sera from patients with KS. Extracts were prepared from BCBL-1 cells (containing KSHV and EBV) and Clone HH514-16 cells (containing only EBV) that were uninduced or treated for 48 hrs with chemical inducing agents, n-butyrate, TPA, or a combination of the two chemicals. Immunoblots prepared from 12% SDS polyacrylamide gels were reacted with a 1:200 dilution of serum from HIV-1 positive patients. FIG. 28A shows serum 01-06 from a patient with KS. FIG. 28B shows serum 01-07 from a patient without KS. FIG. 28C shows serum 04-01 from a patient with KS. FIG. 28D shows serum 01-03 from a patient with KS.
 FIGS. 29A-29F:
 Detection of KSHV lytic cycle antigens by indirect immunofluorescence. BCBL-1 cells were untreated (FIGS. 29A, 29C, and 29E) or treated with n-butyrate (FIGS. 29B, 29D, and 29F) for 48 hrs. Indirect immunofluorescence with a 1:10 dilution of serum from two patients with KS, 04-18 (FIGS. 29A, and 29B) and 04-38 (FIGS. 29E, and 29F) and a serum, 04-37 (FIGS. 29C, and 29D), from a patient without KS.
DETAILED DESCRIPTION OF THE INVENTION
 The following standard abbreviations are used throughout the specification to indicate specific nucleotides:  C=cytosine A=adenosine  T=thymidine G=guanosine
 The term "nucleic acids", as used herein, refers to either DNA or RNA. "Nucleic acid sequence" or "polynucleotide sequence" refers to a single- or double-stranded polymer of deoxyribonucleotide or ribonucleotide bases read from the 5' to the 3' end. It includes both self-replicating plasmids, infectious polymers of DNA or RNA and nonfunctional DNA or RNA.
 By a nucleic acid sequence "homologous to" or "complementary to", it is meant a nucleic acid that selectively hybridizes, duplexes or binds to viral DNA sequences encoding proteins or portions thereof when the DNA sequences encoding the viral protein are present in a human genomic or cDNA library. A DNA sequence which is homologous to a target sequence can include sequences which are shorter or longer than the target sequence so long as they meet the functional test set forth. Hybridization conditions are specified along with the source of the CDNA library.
 Typically, the hybridization is done in a Southern blot protocol using a 0.2×SSC, 0.1% SDS, 65° C. wash. The term "SSC" refers to a citrate-saline solution of 0.15 M sodium chloride and 20 Mm sodium citrate. Solutions are often expressed as multiples or fractions of this concentration. For example, 6×SSC refers to a solution having a sodium chloride and sodium citrate concentration of 6 times this amount or 0.9 M sodium chloride and 120 mM sodium citrate. 0.2×SSC refers to a solution 0.2 times the SSC concentration or 0.03 M sodium chloride and 4 mM sodium citrate.
 The phrase "nucleic acid molecule encoding" refers to a nucleic acid molecule which directs the expression of a specific protein or peptide. The nucleic acid sequences include both the DNA strand sequence that is transcribed into RNA and the RNA sequence that is translated into protein. The nucleic acid molecule include both the full length nucleic acid sequences as well as non-full length sequences derived from the full length protein. It being further understood that the sequence includes the degenerate codons of the native sequence or sequences which may be introduced to provide codon preference in a specific host cell.
 The phrase "expression cassette", refers to nucleotide sequences which are capable of affecting expression of a structural gene in hosts compatible with such sequences. Such cassettes include at least promoters and optionally, transcription termination signals. Additional factors necessary or helpful in effecting expression may also be used as described herein.
 The term "operably linked" as used herein refers to linkage of a promoter upstream from a DNA sequence such that the promoter mediates transcription of the DNA sequence.
 The term "vector", refers to viral expression systems, autonomous self-replicating circular DNA (plasmids), and includes both expression and nonexpression plasmids. Where a recombinant microorganism or cell culture is described as hosting an "expression vector," this includes both extrachromosomal circular DNA and DNA that has been incorporated into the host chromosome(s). Where a vector is being maintained by a host cell, the vector may either be stably replicated by the cells during mitosis as an autonomous structure, or is incorporated within the host's genome.
 The term "plasmid" refers to an autonomous circular DNA molecule capable of replication in a cell, and includes both the expression and nonexpression types. Where a recombinant microorganism or cell culture is described as hosting an "expression plasmid", this includes latent viral DNA integrated into the host chromosome(s). Where a plasmid is being maintained by a host cell, the plasmid is either being stably replicated by the cells during mitosis as an autonomous structure or is incorporated within the host's genome.
 The phrase "recombinant protein" or "recombinantly produced protein" refers to a peptide or protein produced using non-native cells that do not have an endogenous copy of DNA able to express the protein. The cells produce the protein because they have been genetically altered by the introduction of the appropriate nucleic acid sequence. The recombinant protein will not be found in association with proteins and other subcellular components normally associated with the cells producing the protein.
 The following terms are used to describe the sequence relationships between two or more nucleic acid molecules or polynucleotides: "reference sequence", "comparison window", "sequence identity", "percentage of sequence identity", and "substantial identity". A "reference sequence" is a defined sequence used as a basis for a sequence comparison; a reference sequence may be a subset of a larger sequence, for example, as a segment of a full-length cDNA or gene sequence given in a sequence listing or may comprise a complete cDNA or gene sequence.
 Optimal alignment of sequences for aligning a comparison window may be conducted by the local homology algorithm of Smith and Waterman (1981) Adv. Appl. Math. 2:482, by the homology alignment algorithm of Needleman and Wunsch (1970) J. Mol. Biol. 48:443, by the search for similarity method of Pearson and Lipman (1988) Proc. Natl. Acad. Sci. (USA) 85:2444, or by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package Release 7.0, Genetics Computer Group, 575 Science Dr., Madison, Wis.).
 As applied to polypeptides, the terms "substantial identity" or "substantial sequence identity" mean that two peptide sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap which share at least 90 percent sequence identity, preferably at least 95 percent sequence identity, more preferably at least 99 percent sequence identity or more.
 "Percentage amino acid identity" or "percentage amino acid sequence identity" refers to a comparison of the amino acids of two polypeptides which, when optimally aligned, have approximately the designated percentage of the same amino acids. For example, "95% amino acid identity" refers to a comparison of the amino acids of two polypeptides which when optimally aligned have 95% amino acid identity. Preferably, residue positions which are not identical differ by conservative amino acid substitutions. For example, the substitution of amino acids having similar chemical properties such as charge or polarity are not likely to effect the properties of a protein. Examples include glutamine for asparagine or glutamic acid for aspartic acid.
 The phrase "substantially purified" or "isolated" when referring to a herpesvirus peptide or protein, means a chemical composition which is essentially free of other cellular components. It is preferably in a homogeneous state although it can be in either a dry or aqueous solution. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography. A protein which is the predominant species present in a preparation is substantially purified. Generally, a substantially purified or isolated protein will comprise more than 80% of all macromolecular species present in the preparation. Preferably, the protein is purified to represent greater than 90% of all macromolecular species present. More preferably the protein is purified to greater than 95%, and most preferably the protein is purified to essential homogeneity, wherein other macromolecular species are not detected by conventional techniques.
 The phrase "specifically binds to an antibody" or "specifically immunoreactive with", when referring to a protein or peptide, refers to a binding reaction which is determinative of the presence of the herpesvirus of the invention in the presence of a heterogeneous population of proteins and other biologics including viruses other than the herpesvirus. Thus, under designated immunoassay conditions, the specified antibodies bind to the herpesvirus antigens and do not bind in a significant amount to other antigens present in the sample. Specific binding to an antibody under such conditions may require an antibody that is selected for its specificity for a particular protein. For example, antibodies raised to the human herpesvirus immunogen described herein can be selected to obtain antibodies specifically immunoreactive with the herpesvirus proteins and not with other proteins. These antibodies recognize proteins homologous to the human herpesvirus protein. A variety of immunoassay formats may be used to select antibodies specifically immunoreactive with a particular protein. For example, solid-phase ELISA immunoassays are routinely used to select monoclonal antibodies specifically immunoreactive with a protein. See Harlow and Lane  for a description of immunoassay formats and conditions that can be used to determine specific immunoreactivity.
 "Biological sample" as used herein refers to any sample obtained from a living organism or from an organism that has died. Examples of biological samples include body fluids and tissue specimens.
I. Kaposis's Sarcoma (KS)-- Associated Herpesvirus.
 This invention provides an isolated DNA molecule which is at least 30 nucleotides in length and which uniquely defines a herpesvirus associated with Kaposi's sarcoma.
 In one embodiment the isolated DNA molecule comprises at least a portion of the nucleic acid sequence as shown in FIG. 3A (SEQ ID NO: 1). In another embodiment the isolated DNA molecule is a 330 base pair (bp) sequence. In another embodiment the isolated DNA molecule is a 12-50 bp sequence. In another embodiment the isolated DNA molecule is a 30-37 bp sequence.
 In another embodiment the isolated DNA molecule is genomic DNA. In another embodiment the isolated DNA molecule is cDNA. In another embodiment a RNA is derived form the isolated nucleic acid molecule or is capable of hybridizing with the isolated DNA molecule. As used herein "genomic" means both coding and non-coding regions of the isolated nucleic acid molecule.
 Further, the DNA molecule above may be associated with lymphoproliferative diseases including, but not limited to: Hodgkin's disease, non-Hodgkin's lymphoma, lymphatic leukemia, lymphosarcoma, splenomegaly, reticular cell sarcoma, Sezary's syndrome, mycosis fungoides, central nervous system lymphoma, AIDS related central nervous system lymphoma, post-transplant lymphoproliferative disorders, and Burkitt's lymphoma. A lymphoproliferative disorder is characterized as being the uncontrolled clonal or polyclonal expansion of lymphocytes involving lymph nodes, lymphoid tissue and other organs.
 This invention provides an isolated nucleic acid molecule encoding an ORF20 (SEQ ID NOs: 22 and 23), ORF21 (SEQ ID NOs:14 and 15), ORF22 (SEQ ID NOs:16 and 17), ORF23 (SEQ ID NOs:18 and 19), ORF24 (SEQ ID NOs: 20 and 21), ORF25 (SEQ ID NOs: 2 and 3), ORF26 (SEQ ID NOs:24 and 25), ORF27 (SEQ ID NOs:26 and 27), ORF28 (SEQ ID NOs:28 and 29), ORF29A (SEQ ID NOs:30 and 31), ORF29B (SEQ ID NOs:4 and 5), ORF30 (SEQ ID NOs:6 and 7), ORF31 (SEQ ID NOs:8 and 9), ORF32 (SEQ ID NOs:32 and 33), ORF33 (SEQ ID NOs: 10 and 11), ORF34 (SEQ ID NOs: 34 and 35), or ORF35 (SEQ ID NOs:12 AND 13).
 This invention provides an isolated polypeptide encoded by ORF20 (SEQ ID NOs: 22 and 23), ORF21 (SEQ ID NOs:14 and 15), ORF22 (SEQ ID NOs:16 and 17), ORF23 (SEQ ID NOs:18 and 19), ORF24 (SEQ ID NOs: 20 and 21), ORF25 (SEQ ID NOs: 2 and 3), ORF26 (SEQ ID NOs:24 and 25), ORF27 (SEQ ID NOs:26 and 27), ORF28 (SEQ ID NOs:28 and 29), ORF29A (SEQ ID NOs:30 and 31), ORF29B (SEQ ID NOs:4 and 5), ORF30 (SEQ ID NOs:6 and 7), ORF31 (SEQ ID NOs:8 and 9), ORF32 (SEQ ID NOs:32 and 33), ORF33 (SEQ ID NOs: 10 and 11), ORF34 (SEQ ID NOs: 34 and 35), or ORF35 (SEQ ID NOs:12 AND 13).
 For Example, TK is encoded by ORF 21; glycoprotein H (gH) by ORF 22; major capsid protein (MCP) by ORF 25; virion polypeptide (VP23) by ORF 26; and minor capsid protein by ORF 27.
 This invention provides for a replicable vector comprising the isolated DNA molecule of the DNA virus. The vector includes, but is not limited to: a plasmid, cosmid, λ phage or yeast artificial chromosome (YAC) which contains at least a portion of the isolated nucleic acid molecule.
 As an example to obtain these vectors, insert and vector DNA can both be exposed to a restriction enzyme to create complementary ends on both molecules which base pair with each other and are then ligated together with DNA ligase. Alternatively, linkers can be ligated to the insert DNA which correspond to a restriction site in the vector DNA, which is then digested with the restriction enzyme which cuts at that site. Other means are also available and known to an ordinary skilled practitioner.
 Regulatory elements required for expression include promoter or enhancer sequences to bind RNA polymerase and transcription initiation sequences for ribosome binding. For example, a bacterial expression vector includes a promoter such as the lac promoter and for transcription initiation the Shine-Dalgarno sequence and the start codon AUG. Similarly, a eukaryotic expression vector includes a heterologous or homologous promoter for RNA polymerase II, a downstream polyadenylation signal, the start codon AUG, and a termination codon for detachment of the ribosome. Such vectors may be obtained commercially or assembled from the sequences described by methods well-known in the art, for example the methods described above for constructing vectors in general.
 This invention provides a host cell containing the above vector. The host cell may contain the isolated DNA molecule artificially introduced into the host cell. The host cell may be a eukaryotic or bacterial cell (such as E. coli), yeast cells, fungal cells, insect cells and animal cells. Suitable animal cells include, but are not limited to Vero cells, HeLa cells, Cos cells, CV1 cells and various primary mammalian cells.
 This invention provides an isolated herpesvirus associated with Kaposi's sarcoma. In one embodiment the herpesvirus comprises at least a portion of a nucleotide sequence as shown in FIGS. 3A (SEQ ID NO: 1).
 In one embodiment the herpesvirus may be a DNA virus. In another embodiment the herpesvirus may be a Herpesviridae. In another embodiment the herpesvirus may be a gammaherpesvirinae. The classification of the herpesvirus may vary based on the phenotypic or molecular characteristics which are known to those skilled in the art.
 This invention provides an isolated DNA virus wherein the viral DNA is about 270 kb in size, wherein the viral DNA encodes a thymidine kinase, and wherein the viral DNA is capable of selectively hybridizing to a nucleic acid probe selected from the group consisting of SEQ ID NOs: 38-40.
 The KS-associated human herpesvirus of the invention is associated with KS and is involved in the etiology of the disease. The taxonomic classification of the virus has not yet been made and will be based on phenotypic or molecular characteristics known to those of skill in the art. However, the novel KS-associated virus is a DNA virus that appears to be related to the Herpesviridae family and the gammaherpesvirinae subfamily, on the basis of nucleic acid homology.
 A. Sequence Identity of the Viral DNA and its Proteins.
 The human herpesvirus of the invention is not limited to the virus having the specific DNA sequences described herein. The KS-associated human herpesvirus DNA shows substantial sequence identity, as defined above, to the viral DNA sequences described herein. DNA from the human herpesvirus typically selectively hybridizes to one or more of the following three nucleic acid probes:
TABLE-US-00001 Probe 1 (SEQ ID NO: 38) AGCCGAAAGG ATTCCACCAT TGTGCTCGAA TCCAACGGAT TTGACCCCGT GTTCCCCATG GTCGTGCCGC AGCAACTGGG GCACGCTATT CTGCAGCAGC TGTTGGTGTA CCACATCTAC TCCAAAATAT CGGCCGGGGC CCCGGATGAT GTAAATATGG CGGAACTTGA TCTATATACC ACCAATGTGT CATTTATGGG GCGCACATAT CGTCTGGACG TAGACAACAC GGA Probe 2 (SEQ ID NO: 39): GAAATTACCC ACGAGATCGC TTCCCTGCAC ACCGCACTTG GCTACTCATC AGTCATCGCC CCGGCCCACG TGGCCGCCAT AACTACAGAC ATGGGAGTAC ATTGTCAGGA CCTCTTTATG ATTTTCCCAG GGGACGCGTA TCAGGACCGC CAGCTGCATG ACTATATCAA AATGAAAGCG GGCGTGCAAA CCGGCTCACC GGGAAACAGA ATGGATCACG TGGGATACAC TGCTGGGGTT CCTCGCTGCG AGAACCTGCC CGGTTTGAGT CATGGTCAGC TGGCAACCTG CGAGATAATT CCCACGCCGG TCACATCTGA CGTTGCCT Probe 3 (SEQ ID NO: 40): AACACGTCAT GTGCAGGAGT GACATTGTGC CGCGGAGAAA CTCAGACCGC ATCCCGTAAC CACACTGAGT GGGAAAATCT GCTGGCTATG TTTTCTGTGA TTATCTATGC CTTAGATCAC AACTGTCACC CG
 Hybridization of a viral DNA to the nucleic acid probes listed above is determined by using standard nucleic acid hybridization techniques as described herein. In particular, PCR amplification of a viral genome can be carried out using the following three sets of PCR primers:
TABLE-US-00002 1) AGCCGAAAGGATTCCACCAT; (SEQ ID NO: 41) TCCGTGTTGTCTACGTCCAG (SEQ ID NO: 48) 2) GAAATTACCCACGAGATCGC; (SEQ ID NO: 42) AGGCAACGTCAGATGTGA (SEQ ID NO: 49) 3) AACACGTCATGTGCAGGAGTGAC; (SEQ ID NO: 43) CGGGTGACAGTTGTGATCTAAGG (SEQ ID NO: 50)
 In PCR techniques, oligonucleotide primers, as listed above, complementary to the two 3' borders of the DNA region to be amplified are synthesized. The polymerase chain reaction is then carried out using the two primers. See PCR Protocols: A Guide to Methods and Applications . Following PCR amplification, the PCR-amplified regions of a viral DNA can be tested for their ability to hybridize to the three specific nucleic acid probes listed above. Alternatively, hybridization of a viral DNA to the above nucleic acid probes can be performed by a Southern blot procedure without viral DNA amplification and under stringent hybridization conditions as described herein.
 Oligonucleotides for use as probes or PCR primers are chemically synthesized according to the solid phase phosphoramidite triester method first described by Beaucage and Carruthers  using an automated synthesizer, as described in Needham-VanDevanter . Purification of oligonucleotides is by either native acrylamide gel electrophoresis or by anion-exchange HPLC as described in Pearson, J. D. and Regnier, F. E. [75A]. The sequence of the synthetic oligonucleotide can be verified using the chemical degradation method of Maxam, A. M. and Gilbert, W. .
 B. Isolation and Propagation of KS-Inducing Strains of the Human Herpesvirus
 Using conventional methods, the human herpesvirus can be propagated in vitro. For example, standard techniques for growing herpes viruses are described in Ablashi, D. V. . Briefly, PHA stimulated cord blood mononuclear cells, macrophage, neuronal, or glial cell lines are cocultivated with cerebrospinal fluid, plasma, peripheral blood leukocytes, or tissue extracts containing viral infected cells or purified virus. The recipient cells are treated with 5 μg/ml polybrene for 2 hours at 37° C. prior to infection.
 Infected cells are observed by demonstrating morphological changes, as well as being positive for antigens from the human herpesvirus by using monoclonal antibodies immunoreactive with the human herpes virus in an immunofluorescence assay.
 For virus isolation, the virus is either harvested directly from the culture fluid by direct centrifugation, or the infected cells are harvested, homogenized or lysed and the virus is separated from cellular debris and purified by standard methods of isopycnic sucrose density gradient centrifugation.
 One skilled in the art may isolate and propagate the DNA herpesvirus associated with Kaposi's sarcoma (KSHV) employing the following protocol. Long-term establishment of a B lymphoid cell line infected with the KSHV from body-cavity based lymphomas (RCC-1 or HBL-6) is prepared extracting DNA from the Lymphoma tissue using standard techniques [27, 49, 66].
 The KS associated herpesvirus may be isolated from the cell DNA in the following manner. An infected cell line (HBL-6 RCC-1), which can be lysed using standard methods such as hyposomatic shocking and Dounce homogenization, is first pelleted at 2000×g for 10 minutes, the supernatant is removed and centrifuged again at 10,000×g for 15 minutes to remove nuclei and organelles. The supernatant is filtered through a 0.45μ filter and centrifuged again at 100,000×g for 1 hour to pellet the virus. The virus can then be washed and centrifuged again at 100,000×g for 1 hour.
 The DNA is tested for the presence of the KSHV by Southern blotting and PCR using the specific probes as described hereinafter. Fresh lymphoma tissue containing viable infected cells is simultaneously filtered to form a single cell suspension by standard techniques [49, 66]. The cells are separated by standard Ficoll-Plaque centrifugation and lymphocyte layer is removed. The lymphocytes are then placed at >1×106 cells/ml into standard lymphocyte tissue culture medium, such as RMP 1640 supplemented with 10% fetal calf serum. Immortalized lymphocytes containing the KSHV virus are indefinitely grown in the culture media while nonimmortilized cells die during course of prolonged cultivation.
 Further, the virus may be propagated in a new cell line by removing media supernatant containing the virus from a continuously infected cell line at a concentration of >1×106 cells/ml. The media is centrifuged at 2000×g for 10 minutes and filtered through a 0.45μ filter to remove cells. The media is applied in a 1:1 volume with cells growing at >1×106 cells/ml for 48 hours. The cells are washed and pelleted and placed in fresh culture medium, and tested after 14 days of growth.
 RCC-1 and RCC-12F5 were deposited on Oct. 19, 1994 under ATCC Accession No. CRL 11734 and CRL 11735, respectively, pursuant to the Budapest Treaty on the International Deposit of Microorganisms for the Purposes of Patent Procedure with the Patent Culture Depository of the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Md. 20852 U.S.A.
 HBL-6 was deposited on Nov. 18, 1994 under ATCC Accession No. CRL 11762 pursuant to the Budapest Treaty on the International Deposit of Microorganisms for the Purposes of Patent Procedure with the Patent Culture Depository of the American Type Culture Collection, 12301 Parklawn Drive, Rockville, Md. 20852 U.S.A.
 C. Immunological Identity of the Virus
 The KS-associated human herpesvirus can also be described immunologically. KS-associated human herpesviruses are selectively immunoreactive to antisera generated against a defined immunogen such as the viral major capsid protein depicted in Seq. ID No. 12, herein. Immunoreactivity is determined in an immunoassay using a polyclonal antiserum which was raised to the protein which is encoded by the amino acid sequence or nucleic acid sequence of SEQ ID NOs: 18-20. This antiserum is selected to have low crossreactivity against other herpes viruses and any such crossreactivity is removed by immunoabsorption prior to use in the immunoassay.
 In order to produce antisera for use in an immunoassay, the protein which is encoded by the amino acid sequence or nucleic acid of SEQ ID NOs: 18-20 is isolated as described herein. For example, recombinant protein can be produced in a mammalian cell line. An inbred strain of mice such as balb/c is immunized with the protein which is encoded by the amino acid sequence or nucleic acid of SEQ ID NOs: 2-37 using a standard adjuvant, such as Freund's adjuvant, and a standard mouse immunization protocol (see , supra). Alternatively, a synthetic peptide derived from the sequences disclosed herein and conjugated to a carrier protein can be used an immunogen. Polyclonal sera are collected and titered against the immunogen protein in an immunoassay, for example, a solid phase immunoassay with the immunogen immobilized on a solid support. Polyclonal antisera with a titer of 104 or greater are selected and tested for their cross reactivity against other viruses of the gammaherpesvirinae subfamily, particularly human herpes virus types 1-7, by using a standard immunoassay as described in , supra. These other gammaherpesvirinae virus can be isolated by standard techniques for isolation herpes viruses as described herein.
 The ability of the above viruses to compete with the binding of the antisera to the immunogen protein is determined. The percent crossreactivity for other viruses is calculated, using standard calculations. Those antisera with less than 10% crossreactivity with each of the other viruses listed above is selected and pooled. The cross-reacting antibodies are then removed from the pooled antisera by immunoabsorption with the above-listed viruses.
 The immunoabsorbed and pooled antisera are then used in a competitive binding immunoassay procedure as described above to compare an unknown virus preparation to the specific KS herpesvirus preparation described herein and containing the nucleic acid sequence described in SEQ ID NOs: 2-37. In order to make this comparison, the immunogen protein which is encoded by the amino acid sequence or nucleic acid of SEQ ID NOs: 2-37 is the labeled antigen and the virus preparations are each assayed at a wide range of concentrations. The amount of each virus preparation required to inhibit 50% of the binding of the antisera to the labeled immunogen protein is determined. Those viruses that specifically bind to an antibody generated to an immunogen consisting of the protein of SEQ ID NOs: 2-37 are those virus where the amount of virus needed to inhibit 50% of the binding to the protein does not exceed an established amount. This amount is no more than 10 times the amount of the virus that is needed for 50% inhibition for the KS-associated herpesvirus containing the DNA sequence of SEQ ID NO: 1. Thus, the KS-associated herpesviruses of the invention can be defined by immunological comparison to the specific strain of the KS-associated herpesvirus for which nucleic acid sequences are provided herein.
 This invention provides, a nucleic acid molecule of at least 14 nucleotides capable of specifically-hybridizing with the isolated DNA molecule. In one embodiment, the molecule is DNA. In another embodiment, the molecule is RNA. In another embodiment the nucleic acid molecule may be 14-20 nucleotides in length. In another embodiment the nucleic acid molecule may be 16 nucleotides in length.
 This invention provides, a nucleic acid molecule of at least 14 nucleotides capable of specifically hybridizing with a nucleic acid molecule which is complementary to the isolated DNA molecule. In one embodiment, the molecule is DNA. In another embodiment, the molecule is RNA.
 The nucleic acid molecule of at least 14 nucleotides may hybridize with moderate stringency to at least a portion of a nucleic acid molecule with a sequence shown in FIGS. 3A-3F (SEQ ID NOs: 1, and 36-40).
 High stringent hybridization conditions are selected at about 5° C. lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength and pH) at which 50% of the target sequence hybridizes to a perfectly matched probe. Typically, stringent conditions will be those in which the salt concentration is at least about 0.02 molar at pH 7 and the temperature is at least about 60° C. As other factors may significantly affect the stringency of hybridization, including, among others, base composition and size of the complementary strands, the presence of organic solvents, ie. salt or formamide concentration, and the extent of base mismatching, the combination of parameters is more important than the absolute measure of any one. For Example high stringency may be attained for example by overnight hybridization at about 68° C. in a 6×SSC solution, washing at room temperature with 6×SSC solution, followed by washing at about 68° C. in a 6×SSC in a 0.6×SSX solution.
 Hybridization with moderate stringency may be attained for example by: 1) filter pre-hybridizing and hybridizing with a solution of 3× sodium chloride, sodium citrate (SSC), 50% formamide, 0.1M Tris buffer at Ph 7.5, 5×Denhardt's solution; 2.) pre-hybridization at 37° C. for 4 hours; 3) hybridization at 37° C. with amount of labelled probe equal to 3,000,000 cpm total for 16 hours; 4) wash in 2×SSC and 0.1% SDS solution; 5) wash 4× for 1 minute each at room temperature at 4× at 60° C. for 30 minutes each; and 6) dry and expose to film.
 The phrase "selectively hybridizing to" refers to a nucleic acid probe that hybridizes, duplexes or binds only to a particular target DNA or RNA sequence when the target sequences are present in a preparation of total cellular DNA or RNA. By selectively hybridizing it is meant that a probe binds to a given target in a manner that is detectable in a different manner from non-target sequence under high stringency conditions of hybridization, in a different "Complementary" or "target" nucleic acid sequences refer to those nucleic acid sequences which selectively hybridize to a nucleic acid probe. Proper annealing conditions depend, for example, upon a probe's length, base composition, and the number of mismatches and their position on the probe, and must often be determined empirically. For discussions of nucleic acid probe design and annealing conditions, see, for example, Sambrook at al.,  or Ausubel, F., et al., .
 It will be readily understood by those skilled in the art and it is intended here, that when reference is made to particular sequence listings, such reference includes sequences which substantially correspond to its complementary sequence and those described including allowances for minor sequencing errors, single base changes, deletions, substitutions and the like, such that any such sequence variation corresponds to the nucleic acid sequence of the pathogenic organism or disease marker to which the relevant sequence listing relates.
 Nucleic acid probe technology is well known to those skilled in the art who readily appreciate that such probes may vary greatly in length and may be labeled with a detectable label, such as a radioisotope or fluorescent dye, to facilitate detection of the probe. DNA probe molecules may be produced by insertion of a DNA molecule having the full-length or a fragment of the isolated nucleic acid molecule of the DNA virus into suitable vectors, such as plasmids or bacteriophages, followed by transforming into suitable bacterial host cells, replication in the transformed bacterial host cells and harvesting of the DNA probes, using methods well known in the art. Alternatively, probes may be generated chemically from DNA synthesizers.
 DNA virus nucleic acid rearrangements/mutations may be detected by Southern blotting, single stranded conformational polymorphism gel electrophoresis (SSCP), PCR or other DNA based techniques, or for RNA species by Northern blotting, PCR or other RNA-based techniques.
 RNA probes may be generated by inserting the full length or a fragment of the isolated nucleic acid molecule of the DNA virus downstream of a bacteriophage promoter such as T3, T7 or SP6. Large amounts of RNA probe may be produced by incubating the labeled nucleotides with a linearized isolated nucleic acid molecule of the DNA virus or its fragment where it contains an upstream promoter in the presence of the appropriate RNA polymerase.
 As defined herein nucleic acid probes may be DNA or RNA fragments. DNA fragments can be prepared, for example, by digesting plasmid DNA, or by use of PCR, or synthesized by either the phosphoramidite method described by Beaucage and Carruthers, , or by the triester method according to Matteucci, et al., , both incorporated herein by reference. A double stranded fragment may then be obtained, if desired, by annealing the chemically synthesized single strands together under appropriate conditions or by synthesizing the complementary strand using DNA polymerase with an appropriate primer sequence. Where a specific sequence for a nucleic acid probe is given, it is understood that the complementary strand is also identified and included. The complementary strand will work equally well in situations where the target is a double-stranded nucleic acid. It is also understood that when a specific sequence is identified for use a nucleic probe, a subsequence of the listed sequence which is 25 basepairs or more in length is also encompassed for use as a probe.
 The DNA molecules of the subject invention also include DNA molecules coding for polypeptide analogs, fragments or derivatives of antigenic polypeptides which differ from naturally-occurring forms in terms of the identity or location of one or more amino acid residues (deletion analogs containing less than all of the residues specified for the protein, substitution analogs wherein one or more residues specified are replaced by other residues and addition analogs where in one or more amino acid residues is added to a terminal or medial portion of the polypeptides) and which share some or all properties of naturally-occurring forms. These molecules include: the incorporation of codons "preferred" for expression by selected non-mammalian hosts; the provision of sites for cleavage by restriction endonuclease enzymes; and the provision of additional initial, terminal or intermediate DNA sequences that facilitate construction of readily expressed vectors.
 This invention provides for an isolated DNA molecule which encodes at least a portion of a Kaposi's sarcoma associated herpesvirus: virion polypeptide 23, major capsid protein, capsid proteins, thymidine kinase, or tegument protein.
 This invention also provides a method of producing a polypeptide encoded by isolated DNA molecule, which comprises growing the above host vector system under suitable conditions permitting production of the polypeptide and recovering the polypeptide so produced.
 This invention provides an isolated peptide encoded by the isolated DNA molecule associated with Kaposi's sarcoma. In one embodiment the peptide may be a polypeptide. Further, this invention provides a host cell which expresses the polypeptide of isolated DNA molecule.
 In one embodiment the isolated peptide or polypeptide is encoded by at least a portion of an isolated DNA molecule. In another embodiment the isolated peptide or polypeptide is encoded by at least a portion of a nucleic acid molecule with a sequence as set forth in (SEQ ID NOs: 2-37).
 Further, the isolated peptide or polypeptide encoded by the isolated DNA molecule may be linked to a second nucleic acid molecule to form a fusion protein by expression in a suitable host cell. In one embodiment the second nucleic acid molecule encodes beta-galactosidase. Other nucleic acid molecules which are used to form a fusion protein are known to those skilled in the art.
 This invention provides an antibody which specifically binds to the peptide or polypeptide encoded by the isolated DNA molecule. In one embodiment the antibody is a monoclonal antibody. In another embodiment the antibody is a polyclonal antibody.
 The antibody or DNA molecule may be labelled with a detectable marker including, but not limited to: a radioactive label, or a colorimetric, a luminescent, or a fluorescent marker, or gold. Radioactive labels include, but are not limited to: 3H, 14C, 32P, 33P; 35S, 36Cl, 51Cr, 57Co, 59Co, 59Fe, 90Y, 125I, 131I, and 186Re. Fluorescent markers include but are not limited to: fluorescein, rhodamine and auramine. Colorimetric markers include, but are not limited to: biotin, and digoxigenin. Methods of producing the polyclonal or monoclonal antibody are known to those of ordinary skill in the art.
 Further, the antibody or nucleic acid molecule complex may be detected by a second antibody which may be linked to an enzyme, such as alkaline phosphatase or horseradish peroxidase. Other enzymes which may be employed are well known to one of ordinary skill in the art.
 This invention provides a method to select specific regions on the polypeptide encoded by the isolated DNA molecule of the DNA virus to generate antibodies. The protein sequence may be determined from the cDNA sequence. Amino acid sequences may be analyzed by methods well known to those skilled in the art to determine whether they produce hydrophobic or hydrophilic regions in the proteins which they build. In the case of cell membrane proteins, hydrophobic regions are well known to form the part of the protein that is inserted into the lipid bilayer of the cell membrane, while hydrophilic regions are located on the cell surface, in an aqueous environment. Usually, the hydrophilic regions will be more immunogenic than the hydrophobic regions. Therefore the hydrophilic amino acid sequences may be selected and used to generate antibodies specific to polypeptide encoded by the isolated nucleic acid molecule encoding the DNA virus. The selected peptides may be prepared using commercially available machines. As an alternative, DNA, such as a cDNA or a fragment thereof, may be cloned and expressed and the resulting polypeptide recovered and used as an immunogen.
 Polyclonal antibodies against these peptides may be produced by immunizing animals using the selected peptides. Monoclonal antibodies are prepared using hybridoma technology by fusing antibody producing B cells from immunized animals with myeloma cells and selecting the resulting hybridoma cell line producing the desired antibody. Alternatively, monoclonal antibodies may be produced by in vitro techniques known to a person of ordinary skill in the art. These antibodies are useful to detect the expression of polypeptide encoded by the isolated DNA molecule of the DNA virus in living animals, in humans, or in biological tissues or fluids isolated from animals or humans.
 The antibodies raised against the viral strain or peptides may be detectably labelled, utilizing conventional labelling techniques well-known to the art. Thus, the antibodies may be radiolabelled using, for example, radioactive isotopes such as 3H, 125I, 131I, and 35S.
 The antibodies may also be labelled using fluorescent labels, enzyme labels, free radical labels, or bacteriophage labels, using techniques known in the art. Typical fluorescent labels include fluorescein isothiocyanate, rhodamine, phycoerythrin, phycocyanin, alophycocyanin, and Texas Red.
 Since specific enzymes may be coupled to other molecules by covalent links, the possibility also exists that they might be used as labels for the production of tracer materials. Suitable enzymes include alkaline phosphatase, beta-galactosidase, glucose-6-phosphate dehydrogenase, maleate dehydrogenase, and peroxidase. Two principal types of enzyme immunoassay are the enzyme-linked immunosorbent assay (ELISA), and the homogeneous enzyme immunoassay, also known as enzyme-multiplied immunoassay (EMIT, Syva Corporation, Palo Alto, Calif.). In the ELISA system, separation may be achieved, for example, by the use of antibodies coupled to a solid phase. The EMIT system depends on deactivation of the enzyme in the tracer-antibody complex; the activity can thus be measured without the need for a separation step.
 Additionally, chemiluminescent compounds may be used as labels. Typical chemiluminescent compounds include luminol, isoluminol, aromatic acridinium esters, imidazoles, acridinium salts, and oxalate esters. Similarly, bioluminescent compounds may be utilized for labelling, the bioluminescent compounds including luciferin, luciferase, and aequorin.
 Once labeled, the antibody may be employed to identify and quantify immunologic counterparts (antibody or antigenic polypeptide) utilizing techniques well-known to the art.
 A description of a radioimmunoassay (RIA) may be found in Laboratory Techniques in Biochemistry and Molecular Biology , with particular reference to the chapter entitled "An Introduction to Radioimmune Assay and Related Techniques" by Chard, T., incorporated by reference herein.
 A description of general immunometric assays of various types can be found in the following U.S. Pat. Nos. 4,376,110 (David et al.) or 4,098,876 (Piasio).
 A. Assays for Viral Antigens
 In addition to the detection of the causal agent using nucleic acid hybridization technology, one can use immunoassays to detect for the virus, specific peptides, or for antibodies to the virus or peptides. A general overview of the applicable technology is in Harlow and Lane , incorporated by reference herein.
 In one embodiment, antibodies to the human herpesvirus can be used to detect the agent in the sample. In brief, to produce antibodies to the agent or peptides, the sequence being targeted is expressed in transfected cells, preferably bacterial cells, and purified. The product is injected into a mammal capable of producing antibodies. Either monoclonal or polyclonal antibodies (as well as any recombinant antibodies) specific for the gene product can be used in various immunoassays. Such assays include competitive immunoassays, radioimmunoassays, Western blots, ELISA, indirect immunofluorescent assays and the like. For competitive immunoassays, see Harlow and Lane  at pages 567-573 and 584-589.
 Monoclonal antibodies or recombinant antibodies may be obtained by various techniques familiar to those skilled in the art. Briefly, spleen cells or other lymphocytes from an animal immunized with a desired antigen are immortalized, commonly by fusion with a myeloma cell (see, Kohler and Milstein , incorporated herein by reference). Alternative methods of immortalization include transformation with Epstein Barr Virus, oncogenes, or retroviruses, or other methods well known in the art. Colonies arising from single immortalized cells are screened for production of antibodies of the desired specificity and affinity for the antigen, and yield of the monoclonal antibodies produced by such cells may be enhanced by various techniques, including injection into the peritoneal cavity of a vertebrate host. New techniques using recombinant phage antibody expression systems can also be used to generate monoclonal antibodies. See for example: McCafferty, J et al. ; Hoogenboom, H. R. et al. ; and Marks, J. D. et al., .
 Such peptides may be produced by expressing the specific sequence in a recombinantly engineered cell such as bacteria, yeast, filamentous fungal, insect (especially employing baculoviral vectors), and mammalian cells. Those of skill in the art are knowledgeable in the numerous expression systems available for expression of herpes virus protein.
 Briefly, the expression of natural or synthetic nucleic acids encoding viral protein will typically be achieved by operably linking the desired sequence or portion thereof to a promoter (which is either constitutive or inducible), and incorporated into an expression vector. The vectors are suitable for replication or integration in either prokaryotes or eukaryotes. Typical cloning vectors contain antibiotic resistance markers, genes for selection of transformants, inducible or regulatable promoter regions, and translation terminators that are useful for the expression of viral genes.
 Methods for the expression of cloned genes in bacteria are also well known. In general, to obtain high level expression of a cloned gene in a prokaryotic system, it is advisable to construct expression vectors containing a strong promoter to direct mRNA transcription. The inclusion of selection markers in DNA vectors transformed in E. coli is also useful. Examples of such markers include genes specifying resistance to antibiotics. See  supra, for details concerning selection markers and promoters for use in E. coli. Suitable eukaryote hosts may include plant cells, insect cells, mammalian cells, yeast, and filamentous fungi.
 Methods for characterizing naturally processed peptides bound to MHC (major histocompatibility complex) I molecules have been developed. See, Falk et al. , and PCT publication No. WO 92/21033 published Nov. 26, 1992, both of which are incorporated by reference herein. Typically, these methods involve isolation of MHC class I molecules by immunoprecipitation or affinity chromatography from an appropriate cell or cell line. Other methods involve direct amino acid sequencing of the more abundant peptides in various HPLC fractions by known automatic sequencing of peptides eluted from Class I molecules of the B cell type (Jardetzkey, et al. , incorporated by reference herein, and of the human MHC class I molecule, HLA-A2.1 type by mass spectrometry (Hunt, et al. , incorporated by reference herein). See also, Rotzschke and Falk , incorporated by reference herein for a general review of the characterization of naturally processed peptides in MHC class I. Further, Marloes, et al. , incorporated by reference herein, describe how class I binding motifs can be applied to the identification of potential viral immunogenic peptides in vitro.
 The peptides described herein produced by recombinant technology may be purified by standard techniques well known to those of skill in the art. Recombinantly produced viral sequences can be directly expressed or expressed as a fusion protein. The protein is then purified by a combination of cell lysis (e.g., sonication) and affinity chromatography. For fusion products, subsequent digestion of the fusion protein with an appropriate proteolytic enzyme releases the desired peptide.
 The proteins may be purified to substantial purity by standard techniques well known in the art, including selective precipitation with such substances as ammonium sulfate, column chromatography, immunopurification methods, and others. See, for instance, Scopes, R. , incorporated herein by reference.
 B. Serological Tests for the Presence of Antibodies to the Human Herpesvirus.
 This invention further embraces diagnostic kits for detecting the presence of a KS agent in biological samples, such as serum or solid tissue samples, comprising a container containing antibodies to the human herpesvirus, and instructional material for performing the test. Alternatively, inactivated viral particles or peptides or viral proteins derived from the human herpesvirus may be used in a diagnostic kit to detect for antibodies specific to the KS associated human herpesvirus.
 Diagnostic kits for detecting the presence of a KS agent in tissue samples, such as skin samples or samples of other affected tissue, comprising a container containing a nucleic acid sequence specific for the human herpesvirus and instructional material for detecting the KS-associated herpesvirus are also included. A container containing nucleic acid primers to any one of such sequences is optionally included as are antibodies to the human herpesvirus as described herein.
 Antibodies reactive with antigens of the human herpesvirus can also be measured by a variety of immunoassay methods that are similar to the procedures described above for measurement of antigens. For a review of immunological and immunoassay procedures applicable to the measurement of antibodies by immunoassay techniques, see Basic and Clinical Immunology 7th Edition , and , supra.
 In brief, immunoassays to measure antibodies reactive with antigens of the KS-associated human herpesvirus can be either competitive or noncompetitive binding assays. In competitive binding assays, the sample analyte competes with a labeled analyte for specific binding sites on a capture agent bound to a solid surface. Preferably the capture agent is a purified recombinant human herpesvirus protein produced as described above. Other sources of human herpesvirus proteins, including isolated or partially purified naturally occurring protein, may also be used. Noncompetitive assays are typically sandwich assays, in which the sample analyte is bound between two analyte-specific binding reagents. One of the binding agents is used as a capture agent and is bound to a solid surface. The second binding agent is labelled and is used to measure or detect the resultant complex by visual or instrument means. A number of combinations of capture agent and labelled binding agent can be used. A variety of different immunoassay formats, separation techniques and labels can be also be used similar to those described above for the measurement of the human herpesvirus antigens.
 Hemagglutination Inhibition (HI) and Complement Fixation (CF) which are two laboratory tests that can be used to detect infection with human herpesvirus by testing for the presence of antibodies against the virus or antigens of the virus.
 Serological methods can be also be useful when one wishes to detect antibody to a specific variant. For example, one may wish to see how well a vaccine recipient has responded to the new variant.
 Alternatively, one may take serum from a patient to see which variant the patient responds to the best.
 This invention provides an antagonist capable of blocking the expression of the peptide or polypeptide encoded by the isolated DNA molecule. In one embodiment the antagonist is capable of hybridizing with a double stranded DNA molecule. In another embodiment the antagonist is a triplex oligonucleotide capable of hybridizing to the DNA molecule. In another embodiment the triplex oligonucleotide is capable of binding to at least a portion of the isolated DNA molecule with a nucleotide sequence as shown in FIG. 3A-3F (SEQ ID NOs: 1, and 36-40).
 This invention provides an antisense molecule capable of hybridizing to the isolated DNA molecule. In one embodiment the antisense molecule is DNA. In another embodiment the antisense molecule is RNA.
 The antisense molecule may be DNA or RNA or variants thereof (i.e. DNA or RNA with a protein backbone). The present invention extends to the preparation of antisense nucleotides and ribozymes that may be used to interfere with the expression of the receptor recognition proteins at the translation of a specific mRNA, either by masking that mRNA with an antisense nucleic acid or cleaving it with a ribozyme.
 Antisense nucleic acids are DNA or RNA molecules that are complementary to at least a portion of a specific mRNA molecule. In the cell, they hybridize to that mRNA, forming a double stranded molecule. The cell does not translate an mRNA in this double-stranded 35 form. Therefore, antisense nucleic acids interfere with the expression of mRNA into protein. Oligomers of about fifteen nucleotides and molecules that hybridize to the AUG initiation codon are particularly efficient, since they are easy to synthesize and are likely to pose fewer problems than larger molecules upon introduction to cells.
 This invention provides a transgenic nonhuman mammal which comprises at least a portion of the isolated DNA molecule introduced into the mammal at an embryonic stage. Methods of producing a transgenic nonhuman mammal are known to those skilled in the art.
 This invention provides a cell line containing the isolated KS associated herpesvirus of the subject invention. In one embodiment the isolated DNA molecule is artificially introduced into the cell. Cell lines include, but are not limited to: fibroblasts, such as HFF, NIH/3T3; Epithelial cells, such as 5637; lymphocytes, such as FCB; T-cells, such as CCRF-CEM (ATCC CCL 119); B-cells, such as BJAB and Raji (ATCC CCL 86); and myeloid cells such as K562 (ATCC CCL 243); Vero cells and carcinoma cells. Methods of producing such cell lines are known to those skilled in the art. In one embodiment the isolated KS associated herpesvirus is introduced into a RCC-1 cell line.
III. In Vitro Diagnostic Assays for the Detection of KS
 This invention provides a method of diagnosing Kaposi's sarcoma in a subject which comprises: (a) obtaining a nucleic acid molecule from a tumor lesion of the subject: (b) contacting the nucleic acid molecule with a labelled nucleic acid molecule of at least 15 nucleotides capable of specifically hybridizing with the isolated DNA, under hybridizing conditions; and (c) determining the presence of the nucleic acid molecule hybridized, the presence of which is indicative of Kaposi's sarcoma in the subject, thereby diagnosing Kaposi's sarcoma in the subject.
 In one embodiment the DNA molecule from the tumor lesion is amplified before step (b). In another embodiment PCR is employed to amplify the nucleic acid molecule. Methods of amplifying nucleic acid molecules are known to those skilled in the art.
 A person of ordinary skill in the art will be able to obtain appropriate DNA sample for diagnosing Kaposi's sarcoma in the subject. The DNA sample obtained by the above described method may be cleaved by restriction enzyme. The uses of restriction enzymes to cleave DNA and the conditions to perform such cleavage are well-known in the art.
 In the above described methods, a size fractionation may be employed which is effected by a polyacrylamide gel. In one embodiment, the size fractionation is effected by an agarose gel. Further, transferring the DNA fragments into a solid matrix may be employed before a hybridization step. One example of such solid matrix is nitrocellulose paper.
 This invention provides a method of diagnosing Kaposi's sarcoma in a subject which comprises: (a) obtaining a nucleic acid molecule from a suitable bodily fluid of the subject; (b) contacting the nucleic acid molecule with a labelled nucleic acid molecules of at least 15 nucleotides capable of specifically hybridizing with the isolated DNA, under hybridizing conditions; and (c) determining the presence of the nucleic acid molecule hybridized, the presence of which is indicative of Kaposi's sarcoma in the subject, thereby diagnosing Kaposi's sarcoma in the subject.
 This invention provides a method of diagnosing a DNA virus in a subject, which comprises (a) obtaining a suitable bodily fluid sample from the subject, (b) contacting the suitable bodily fluid of the subject to a support having already bound thereto a Kaposi's sarcoma antibody, so as to bind the Kaposi's sarcoma antibody to a specific Kaposi's sarcoma antigen, (c) removing unbound bodily fluid from the support, and (d) determining the level of Kaposi's sarcoma antibody bound by the Kaposi's sarcoma antigen, thereby diagnosing the subject for Kaposi's sarcoma.
 This invention provides a method of diagnosing Kaposi's sarcoma in a subject, which comprises (a) obtaining a suitable bodily fluid sample from the subject, (b) contacting the suitable bodily fluid of the subject to a support having already bound thereto a Kaposi's sarcoma antigen, so as to bind Kaposi's sarcoma antigen to a specific Kaposi's sarcoma antibody, (c) removing unbound bodily fluid from the support, and (d) determining the level of the Kaposi's sarcoma antigen bound by the Kaposi's sarcoma antibody, thereby diagnosing Kaposi's sarcoma.
 This invention provides a method of detecting expression of a DNA virus associated with Kaposi's sarcoma in a cell which comprises obtaining total cDNA obtained from the cell, contacting the cDNA so obtained with a labelled DNA molecule under hybridizing conditions, determining the presence of cDNA hybridized to the molecule, and thereby detecting the expression of the DNA virus. In one embodiment mRNA is obtained from the cell to detect expression of the DNA virus.
 The suitable bodily fluid sample is any bodily fluid sample which would contain Kaposi's sarcoma antibody, antigen or fragments thereof. A suitable bodily fluid includes, but is not limited to: serum, plasma, cerebrospinal fluid, lymphocytes, urine, transudates, or exudates. In the preferred embodiment, the suitable bodily fluid sample is serum or plasma. In addition, the bodily fluid sample may be cells from bone marrow, or a supernatant from a cell culture. Methods of obtaining a suitable bodily fluid sample from a subject are known to those skilled in the art. Methods of determining the level of antibody or antigen include, but are not limited to: ELISA, IFA, and Western blotting. Other methods are known to those skilled in the art. Further, a subject infected with a DNA virus associated with Kaposi's sarcoma may be diagnosed with the above described methods.
 The detection of the human herpesvirus and the detection of virus-associated KS are essentially identical processes. The basic principle is to detect the virus using specific ligands that bind to the virus but not to other proteins or nucleic acids in a normal human cell or its environs. The ligands can either be nucleic acid or antibodies. The ligands can be naturally occurring or genetically or physically modified such as nucleic acids with non-natural or antibody derivatives, i.e., Fab or chimeric antibodies. Serological tests for detection of antibodies to the virus may also be performed by using protein antigens obtained from the human herpesvirus, and described herein.
 Samples can be taken from patients with KS or from patients at risk for KS, such as AIDS patients. Typically the samples are taken from blood (cells, serum and/or plasma) or from solid tissue samples such as skin lesions. The most accurate diagnosis for KS will occur if elevated titers of the virus are detected in the blood or in involved lesions. KS may also be indicated if antibodies to the virus are detected and if other diagnostic factors for KS is present.
 A. Nucleic Acid Assays.
 The diagnostic assays of the invention can be nucleic acid assays such as nucleic acid hybridization assays and assays which detect amplification of specific nucleic acid to detect for a nucleic acid sequence of the human herpesvirus described herein.
 Accepted means for conducting hybridization assays are known and general overviews of the technology can be had from a review of Nucleic Acid Hybridization: A Practical Approach ; Hybridization of Nucleic Acids Immobilized on Solid Supports ; Analytical Biochemistry  and Innis et al., PCR Protocols , supra, all of which are incorporated by reference herein.
 If PCR is used in conjunction with nucleic acid hybridization, primers are designed to target a specific portion of the nucleic acid of the herpesvirus. For example, the primers set forth in SEQ ID NOs: 38-40 may be used to target detection of regions of the herpesvirus genome encoding ORF 25 homologue-ORF 32 homologue. From the information provided herein, those of skill in the art will be able to select appropriate specific primers.
 Target specific probes may be used in the nucleic acid hybridization diagnostic assays for KS. The probes are specific for or complementary to the target of interest. For precise allelic differentiations, the probes should be about 14 nucleotides long and preferably about 20-30 nucleotides. For more general detection of the human herpesvirus of the invention, nucleic acid probes are about 50 to about 1000 nucleotides, most preferably about 200 to about 400 nucleotides.
 A sequence is "specific" for a target organism of interest if it includes a nucleic acid sequence which when detected is determinative of the presence of the organism in the presence of a heterogeneous population of proteins and other biologics. A specific nucleic acid probe is targeted to that portion of the sequence which is determinative of the organism and will not hybridize to other sequences especially those of the host where a pathogen is being detected.
 The specific nucleic acid probe can be RNA or DNA polynucleotide or oligonucleotide, or their analogs. The probes may be single or double stranded nucleotides. The probes of the invention may be synthesized enzymatically, using methods well known in the art (e.g., nick translation, primer extension, reverse transcription, the polymerase chain reaction, and others) or chemically (e.g., by methods such as the phosphoramidite method described by Beaucage and Carruthers , or by the triester method according to Matteucci, et al. , both incorporated herein by reference).
 The probe must be of sufficient length to be able to form a stable duplex with its target nucleic acid in the sample, i.e., at least about 14 nucleotides, and may be longer (e.g., at least about 50 or 100 bases in length). Often the probe will be more than about 100 bases in length. For example, when probe is prepared by nick-translation of DNA in the presence of labeled nucleotides the average probe length may be about 100-600 bases.
 As noted above, the probe will be capable of specific hybridization to a specific KS-associated herpes virus nucleic acid. Such "specific hybridization" occurs when a probe hybridizes to a target nucleic acid, as evidenced by a detectable signal, under conditions in which the probe does not hybridize to other nucleic acids (e.g., animal cell or other bacterial nucleic acids) present in the sample. A variety of factors including the length and base composition of the probe, the extent of base mismatching between the probe and the target nucleic acid, the presence of salt and organic solvents, probe concentration, and the temperature affect hybridization, and optimal hybridization conditions must often be determined empirically. For discussions of nucleic acid probe design and annealing conditions, see, for example, , supra, Ausubel, F., et al.  [ hereinafter referred to as Sambrook], Methods in Enzymology  or Hybridization with Nucleic Acid Probes  all of which are incorporated herein by reference.
 Usually, at least a part of the probe will have considerable sequence identity with the target nucleic acid. Although the extent of the sequence identity required for specific hybridization will depend on the length of the probe and the hybridization conditions, the probe will usually have at least 70% identity to the target nucleic acid, more usually at least 80% identity, still more usually at least 90% identity and most usually at least 95% or 100% identity.
 A probe can be identified as capable of hybridizing specifically to its target nucleic acid by hybridizing the probe to a sample treated according the protocol of this invention where the sample contains both target virus and animal cells (e.g., nerve cells). A probe is specific if the probe's characteristic signal is associated with the herpesvirus DNA in the sample and not generally with the DNA of the host cells and non-biological materials (e.g., substrate) in a sample.
 The following stringent hybridization and washing conditions will be adequate to distinguish a specific probe (e.g., a fluorescently labeled DNA probe) from a probe that is not specific: incubation of the probe with the sample for 12 hours at 37° C. in a solution containing denatured probe, 50% formamide, 2×SSC, and 0.1% (w/v) dextran sulfate, followed by washing in 1×SSC at 70° C. for 5 minutes; 2×SSC at 37° C. for 5 minutes; 0.2×SSC at room temperature for 5 minutes, and H2O at room temperature for 5 minutes. Those of skill will be aware that it will often be advantageous in nucleic acid hybridizations (i.e., in situ, Southern, or other) to include detergents (e.g., sodium dodecyl sulfate), chelating agents (e.g., EDTA) or other reagents (e.g., buffers, Denhardt's solution, dextran sulfate) in the hybridization or wash solutions. To test the specificity of the virus specific probes, the probes can be tested on host cells containing the KS-associated herpesvirus and compared with the results from cells containing non-KS-associated virus.
 It will be apparent to those of ordinary skill in the art that a convenient method for determining whether a probe is specific for a KS-associated viral nucleic acid utilizes a Southern blot (or Dot blot) using DNA prepared from one or more KS-associated human herpesviruses of the invention. Briefly, to identify a target specific probe DNA is isolated from the virus. Test DNA either viral or cellular is transferred to a solid (e.g., charged nylon) matrix. The probes are labelled following conventional methods. Following denaturation and/or prehybridization steps known in the art, the probe is hybridized to the immobilized DNAs under stringent conditions. Stringent hybridization conditions will depend on the probe used and can be estimated from the calculated Tm (melting temperature) of the hybridized probe (see, e.g., Sambrook for a description of calculation of the Tm). For radioactively-labeled DNA or RNA probes an example of stringent hybridization conditions is hybridization in a solution containing denatured probe and 5×SSC at 65° C. for 8-24 hours followed by washes in 0.1×SSC, 0.1% SDS (sodium dodecyl sulfate) at 50-65° C. In general, the temperature and salt concentration are chosen so that the post hybridization wash occurs at a temperature that is about 5° C. below the TM of the hybrid. Thus for a particular salt concentration the temperature may be selected that is 5° C. below the TM or conversely, for a particular temperature, the salt concentration is chosen to provide a TM for the hybrid that is 5° C. warmer than the wash temperature. Following stringent hybridization and washing, a probe that hybridizes to the KS-associated viral DNA but not to the non-KS associated viral DNA, as evidenced by the presence of a signal associated with the appropriate target and the absence of a signal from the non-target nucleic acids, is identified as specific for the KS associated virus. It is further appreciated that in determining probe specificity and in utilizing the method of this invention to detect KS-associated herpesvirus, a certain amount of background signal is typical and can easily be distinguished by one of skill from a specific signal. Two fold signal over background is acceptable.
 A preferred method for detecting the KS-associated herpesvirus is the use of PCR and/or dot blot hybridization. The presence or absence of an KS agent for detection or prognosis, or risk assessment for KS includes Southern transfers, solution hybridization or non-radioactive detection systems, all of which are well known to those of skill in the art. Hybridization is carried out using probes. Visualization of the hybridized portions allows the qualitative determination of the presence or absence of the causal agent.
 Similarly, a Northern transfer may be used for the detection of message in samples of RNA or reverse transcriptase PCR and cDNA can be detected by methods described above. This procedure is also well known in the art. See  incorporated by reference herein.
 An alternative means for determining the presence of the human herpesvirus is in situ hybridization, or more recently, in situ polymerase chain reaction. In situ PCR is described in Neuvo et al. , Intracellular localization of polymerase chain reaction (PCR)-amplified Hepatitis C cDNA; Bagasra et al. , Detection of Human Immunodeficiency virus type 1 provirus in mononuclear cells by in situ polymerase chain reaction; and Heniford et al. , Variation in cellular EGF receptor mRNA expression demonstrated by in situ reverse transcriptase polymerase chain reaction. In situ hybridization assays are well known and are generally described in Methods Enzymol.  incorporated by reference herein. In an in situ hybridization, cells are fixed to a solid support, typically a glass slide. The cells are then contacted with a hybridization solution at a moderate temperature to permit annealing of target-specific probes that are labelled. The probes are preferably labelled with radioisotopes or fluorescent reporters.
 The above described probes are also useful for in-situ hybridization or in order to locate tissues which express this gene, or for other hybridization assays for the presence of this gene or its mRNA in various biological tissues. In-situ hybridization is a sensitive localization method which is not dependent on expression of antigens or native vs. denatured conditions.
 Oligonucleotide (oligo) probes, synthetic oligonucleotide probes or riboprobes made from KSHV phagemids/plasmids, are relatively homogeneous reagents and successful hybridization conditions in tissue sections is readily transferable from one probe to another. Commercially synthesized oligonucleotide probes are prepared against the identified genes. These probes are chosen for length (45-65 mers), high G-C content (50-70%) and are screened for uniqueness against other viral sequences in GenBank.
 Oligonucleotides are 3' end-labeled with [α-35S]dATP to specific activities in the range of 1×1010 dpm/ug using terminal deoxynucleotidyl transferase. Unincorporated labeled nucleotides are removed from the oligo probe by centrifugation through a Sephadex G-25 column or by elution from a Waters Sep Pak C-18 column.
 KS tissue embedded in OCT compound and snap frozen in freezing isopentane cooled with dry ice is cut at 6 μm intervals and thawed onto 3-aminopropyltriethoxysilane treated slides and allowed to air dry. The slides are then be fixed in 4% freshly prepared paraformaldehyde, rinsed in water. Formalin-fixed, paraffin embedded KS tissues cut at 6 μm and baked onto glass slides can also be used. The sections are then deparaffinized in xylenes and rehydrated through graded alcohols. Prehybridization in 20 mM Tris Ph 7.5, 0.02% Denhardt's solution, 10% dextran sulfate for 30 min at 37° C. is followed by hybridization overnight in a solution of 50% formamide (v/v), 10% dextran sulfate (w/v), 20 mM sodium phosphate (Ph 7.4), 3×SSC, 1×Denhardt's solution, 100 ug/ml salmon sperm DNA, 125 ug/ml yeast tRNA and the oligo probe (106 cpm/ml) at 42° C. overnight. The slides are washed twice with 2×SSC and twice with 1×SSC for 15 minutes each at room temperature and visualized by autoradiography. Briefly, sections are dehydrated through graded alcohols containing 0.3M ammonium acetate and air dried. The slides are dipped in Kodak NTB2 emulsion, exposed for days to weeks, developed, and counterstained with hematoxylin and eoxin. Alternative immunohistochemical protocols may be employed which are known to those skilled in the art.
IV. Treatment of Human Herpesvirus-Induced KS
 This invention provides a method of treating a subject with Kaposi's sarcoma, comprising administering to the subject an effective amount of the antisense molecule capable of hybridizing to the isolated DNA molecule under conditions such that the antisense molecule selectively enters a tumor cell of the subject, so as to treat the subject.
 This invention provides a method for treating a subject with Kaposi's sarcoma (KS) comprising administering to the subject having a human herpesvirus-associated KS a pharmaceutically effective amount of an antiviral agent in a pharmaceutically acceptable carrier, wherein the agent is effective to treat the subject with KS-associated human herpes virus.
 Further, this invention provides a method of prophylaxis or treatment for Kaposi's sarcoma (KS) by administering to a patient at risk for KS, an antibody that binds to the human herpesvirus in a pharmaceutically acceptable carrier. In one embodiment the antiviral drug is used to treat a subject with the DNA herpesvirus of the subject invention.
 The use of combinations of antiviral drugs and sequential treatments are useful for treatment of herpesvirus infections and will also be useful for the treatment of herpesvirus-induced KS. For example, Snoeck at al. , found additive or synergistic effects against CMV when combining antiherpes drugs (e.g., combinations of zidovudine [3'-azido-3'-deoxythymidine, AZT] with HPMPC, ganciclovir, foscarnet or acyclovir or of HPMPC with other antivirals). Similarly, in treatment of cytomegalovirus retinitis, induction with ganciclovir followed by maintenance with foscarnet has been suggested as a way to maximize efficacy while minimizing the adverse side effects of either treatment alone. An anti-herpetic composition that contains acyclovir and, e.g., 2-acetylpyridine-5-((2-pyridylamino)thiocarbonyl)-thiocarbonohydrazone is described in U.S. Pat. No. 5,175,165 (assigned to Burroughs Wellcome Co.). Combinations of TS-inhibitors and viral TK-inhibitors in antiherpetic medicines are disclosed in U.S. Pat. No. 5,137,724, assigned to Stichting Rega VZW. A synergistic inhibitory effect on EBV replication using certain ratios of combinations of HPMPC with AZT was reported by Lin et al. .
 U.S. Pat. Nos. 5,164,395 and 5,021,437 (Blumenkopf; Burroughs Wellcome) describe the use of a ribonucleotide reductase inhibitor (an acetylpyridine derivative) for treatment of herpes infections, including the use of the acetylpyridine derivative in combination with acyclovir. U.S. Pat. No. 5,137,724 (Balzari et al. ) describes the use of thymilydate synthase inhibitors (e.g., 5-fluoro-uracil and 5-fluoro-2'-deoxyuridine) in combination with compounds having viral thymidine kinase inhibiting activity.
 With the discovery of a disease causal agent for KS now identified, effective therapeutic or prophalactic protocols to alleviate or prevent the symptoms of herpes virus-associated KS can be formulated. Due to the viral nature of the disease, antiviral agents have application here for treatment, such as interferons, nucleoside analogues, ribavirin, amantadine, and pyrophosphate analogues of phosphonoacetic acid (foscarnet) (reviewed in Gorbach, S. L., et al. ) and the like. Immunological therapy will also be effective in many cases to manage and alleviate symptoms caused by the disease agents described here. Antiviral agents include agents or compositions that directly bind to viral products and interfere with disease progress; and, excludes agents that do not impact directly on viral multiplication or viral titer. Antiviral agents do not include immunoregulatory agents that do not directly affect viral titer or bind to viral products. Antiviral agents are effective if they inactivate the virus, otherwise inhibit its infectivity or multiplication, or alleviate the symptoms of KS.
 A. Antiviral Agents.
 The antiherpesvirus agents that will be useful for treating virus-induced KS can be grouped into broad classes based on their presumed modes of action. These classes include agents that act (i) by inhibition of viral DNA polymerase, (ii) by targeting other viral enzymes and proteins, (iii) by miscellaneous or incompletely understood mechanisms, or (iv) by binding a target nucleic acid (i.e., inhibitory nucleic acid therapeutics). Antiviral agents may also be used in combination (i.e., together or sequentially) to achieve synergistic or additive effects or other benefits.
 Although it is convenient to group antiviral agents by their supposed mechanism of action, the applicants do not intend to be bound by any particular mechanism of antiviral action. Moreover, it will be understood by those of skill that an agent may act on more than one target in a virus or virus-infected cell or through more than one mechanism.
 i) Inhibitors of Viral DNA Polymerase
 Many antiherpesvirus agents in clinical use or in development today are nucleoside analogs believed to act through inhibition of viral DNA replication, especially through inhibition of viral DNA polymerase. These nucleoside analogs act as alternative substrates for the viral DNA polymerase or as competitive inhibitors of DNA polymerase substrates. Usually these agents are preferentially phosphorylated by viral thymidine kinase (TK), if one is present, and/or have higher affinity for viral DNA polymerase than for the cellular DNA polymerases, resulting in selective antiviral activity. Where a nucleoside analogue is incorporated into the viral DNA, viral activity or reproduction may be affected in a variety of ways. For example, the analogue may act as a chain terminator, cause increased lability (e.g., susceptibility to breakage) of analogue-containing DNA, and/or impair the ability of the substituted DNA to act as template for transcription or replication (see, e.g., Balzarini et al. ).
 It will be known to one of skill that, like many drugs, many of the agents useful for treatment of herpes virus infections are modified (i.e., "activated") by the host, host cell, or virus-infected host cell metabolic enzymes. For example, acyclovir is triphosphorylated to its active form, with the first phosphorylation being carried out by the herpes virus thymidine kinase, when present. Other examples are the reported conversion of the compound HOE 602 to ganciclovir in a three-step metabolic pathway (Winkler et al. ) and the phosphorylation of ganciclovir to its active form by, e.g., a CMV nucleotide kinase. It will be apparent to one of skill that the specific metabolic capabilities of a virus can affect the sensitivity of that virus to specific drugs, and is one factor in the choice of an antiviral drug. The mechanism of action of certain anti-herpesvirus agents is discussed in De Clercq  and in other references cited supra and infra, all of which are incorporated by reference herein.
 Anti-herpesvirus medications suitable for treating viral induced KS include, but are not limited to, nucleoside analogs including acyclic nucleoside phosphonate analogs (e.g., phosphonylmethoxyalkylpurines and -pyrimidines), and cyclic nucleoside analogs. These include drugs such as: vidarabine (9-β-D-arabinofuranosyladenine; adenine arabinoside, ara-A, Vira-A, Parke-Davis); 1-β-D-arabinofuranosyluracil (ara-U); 1-β-D-arabinofuranosyl-cytosine (ara-C); HPMPC [(S)-1-β-[3-hydroxy-2-(phosphonylmethoxy)propyl]cytosine (e.g., GS 504 Gilead Science)] and its cyclic form (cHPMPC); HPMPA [(S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)adenine] and its cyclic form (cHPMPA); (S)--HPMPDAP [(S)-9-(3-hydroxy-2-phosphonylmethoxypropyl)-2,6-diaminopurine]; PMEDAP [9-(2-phosphonyl-methoxyethyl)-2,6-diaminopurine]; HOE 602 [2-amino-9-(1,3-bis(isopropoxy)-2-propoxymethyl)purine]; PMEA [9-(2-phosphonylmethoxyethyl)adenine]; bromovinyl-deoxyuridine (Burns and Sandford. ); 1-β-D-arabinofuranosyl-E-5-(2-bromovinyl)-uridine or -2'-deoxyuridine; BVaraU (1-β-D-arabinofuranosyl-E-5-(2-bromovinyl)-uracil, brovavir, Bristol-Myers Squibb, Yamsa Shoyu); BVDU [(E)-5-(2-bromovinyl)-2'-deoxyuridine, brivudin, e.g., Helpin] and its carbocyclic analogue (in which the sugar moiety is replaced by a cyclopentane ring); IVDU [(E)-5-(2-iodovinyl)-2'-deoxyuridine) and its carbocyclic analogue, C-IVDU (Balzarini et al. )]; and 5-mercutithio analogs of 2'-deoxyuridine (Holliday, J., and Williams, M. V. ); acyclovir [9-([2-hydroxyethoxy]methyl)guanine; e.g., Zovirax (Burroughs Wellcome)]; penciclovir (9-[4-hydroxy-2-(hydroxymethyl)butyl]-guanine); ganciclovir [(9-[1,3-dihydroxy-2 propoxymethyl]-guanine) e.g., Cymevene, Cytovene (Syntex), DHPG (Stals et al. ]; isopropylether derivatives of ganciclovir (see, e.g., Winkelmann et al., ); cygalovir; famciclovir [2-amino-9-(4-acetoxy-3-(acetoxymethyl)but-1-yl)purine (Smithkline Beecham)]; valacyclovir (Burroughs Wellcome); desciclovir [(2-amino-9-(2-ethoxymethyl)purine)] and 2-amino-9-(2-hydroxyethoxymethyl)-9H-purine, prodrugs of acyclovir]; CDG (carbocyclic 2'-deoxyguanosine); and purine nucleosides with the pentafuranosyl ring replaced by a cyclo butane ring (e.g., cyclobut-A [(+-)-9-[1β,2α,3β)-2,3-bis(hydroxymethyl)-1-cyclobutyl]ad- enine], cyclobut-G [(+-)-9-[(1β,2α,3β)-2,3-bis(hydroxymethyl)-1-cyclobutyl]g- uanine], BHCG [(R)-(1α,2β,1α)-9-(2,3-bis(hydroxymethyl)cyclobutyl]guan- ine], and an active isomer of racemic BHCG, SQ 34,514 [1R-1α,2β,3α)-2-amino-9-[2,3-bis(hydroxymethyl)cyclobuty- l]-6H-purin-6-one (see, Braitman et al. (1991) ]. Certain of these antiherpesviral agents are discussed in Gorach at al. ; Saunders et al., ; Yamanaka et al., ; Greenspan et al. , all of which are incorporated by reference herein.
 Triciribine and triciribine monophosphate are potent inhibitors against herpes viruses. (Ickes et al. , incorporated by reference herein), HIV-1 and HIV-2 (Kucera et al. , incorporated by reference herein) and are additional nucleoside analogs that may be used to treat KS. An exemplary protocol for these agents is an intravenous injection of about 0.35 mg/meter2 (0.7 mg/kg) once weekly or every other week for at least two doses, preferably up to about four to eight weeks.
 Acyclovir and ganciclovir are of interest because of their accepted use in clinical settings. Acyclovir, an acyclic analogue of guanine, is phosphorylated by a herpesvirus thymidine kinase and undergoes further phosphorylation to be incorporated as a chain terminator by the viral DNA polymerase during viral replication. It has therapeutic activity against a broad range of herpesviruses, Herpes simplex Types 1 and 2, Varicella-Zoster, Cytomegalovirus, and Epstein-Barr Virus, and is used to treat disease such as herpes encephalitis, neonatal herpesvirus infections, chickenpox in immunocompromised hosts, herpes zoster recurrences, CMV retinitis, EBV infections, chronic fatigue syndrome, and hairy leukoplakia in AIDS patients. Exemplary intravenous dosages or oral dosages are 250 mg/kg/m2 body surface area, every 8 hours for 7 days, or maintenance doses of 200-400 mg IV or orally twice a day to suppress recurrence. Ganciclovir has been shown to be more active than acyclovir against some herpesviruses. See, e.g., Oren and Soble . Treatment protocols for ganciclovir are 5 mg/kg twice a day IV or 2.5 mg/kg three times a day for 10-14 days. Maintenance doses are 5-6 mg/kg for 5-7 days.
 Also of interest is HPMPC. HPMPC is reported to be more active than either acyclovir or ganciclovir in the chemotherapy and prophylaxis of various HSV-1, HSV-2, TK-HSV, VZV or CMV infections in animal models (, supra).
 Nucleoside analogs such as BVaraU are potent inhibitors of HSV-1, EBV, and VZV that have greater activity than acyclovir in animal models of encephalitis. FIAC (fluoroidoarbinosyl cytosine) and its related fluoroethyl and iodo compounds (e.g., FEAU, FIAU) have potent selective activity against herpesviruses, and HPMPA ((S)-1-([3-hydroxy-2-phosphorylmethoxy]propyl)adenine) has been demonstrated to be more potent against HSV and CMV than acyclovir or ganciclovir and are of choice in advanced cases of KS. Cladribine (2-chlorodeoxyadenosine) is another nucleoside analogue known as a highly specific antilymphocyte agent (i.e., a immunosuppressive drug).
 Other useful antiviral agents include: 5-thien-2-yl-2'-deoxyuridine derivatives, e.g., BTDU [5-5(5-bromothien-2-yl)-2'-deoxyuridine] and CTDU [b-(5-chlorothien-2-yl)-2'-deoxyuridine]: and OXT-A [9-(2-deoxy-2-hydroxymethyl-β-D-erythro-oxetanosyl)adenine] and OXT-G [9-(2-deoxy-2-hydroxymethyl-O-D-erythro-oxetanosyl)guanine]. Although OXT-G is believed to act by inhibiting viral DNA synthesis its mechanism of action has not yet been elucidated. These and other compounds are described in Andrei et al.  which is incorporated by reference herein. Additional antiviral purine derivatives useful in treating herpesvirus infections are disclosed in U.S. Pat. No. 5,108,994 (assigned to Beecham Group P.L.C.). 6-Methoxypurine arabinoside (ara-M, Burroughs Wellcome) is a potent inhibitor of varicella-zoster virus, and will be useful for treatment of KS.
 Certain thymidine analogs [e.g., idoxuridine (5-ido-2'-deoxyuridine)] and trifluorothymidine) have antiherpes viral activity, but due to their systemic toxicity, are largely used for topical herpesviral infections, including HSV stromal keratitis and uveitis, and are not preferred here unless other options are ruled out.
 Other useful antiviral agents that have demonstrated antiherpes viral activity include foscarnet sodium (trisodium phosphonoformate, PFA, Foscavir (Astra)) and phosphonoacetic acid (PAA). Foscarnet is an inorganic pyrophosphate analogue that acts by competitively blocking the pyrophosphate-binding site of DNA polymerase. These agents which block DNA polymerase directly without processing by viral thymidine kinase. Foscarnet is reported to be less toxic than PAA.
 ii) Agents that Target Viral Proteins Other than DNA Polymerase or Other Viral Functions.
 Although applicants do not intend to be bound by a particular mechanism of antiviral action, the antiherpes-virus agents described above are believed to act through inhibition of viral DNA polymerase. However, viral replication requires not only the replication of the viral nucleic acid but also the production of viral proteins and other essential components. Accordingly, the present invention contemplates treatment of KS by the inhibition of viral proliferation by targeting viral proteins other than DNA polymerase (e.g., by inhibition of their synthesis or activity, or destruction of viral proteins after their synthesis). For example, administration of agents that inhibit a viral serine protease, e.g., such as one important in development of the viral capsid will be useful in treatment of viral induced KS.
 Other viral enzyme targets include: OMP decarboxylase inhibitors (a target of, e.g., parazofurin), CTP synthetase inhibitors (targets of, e.g., cyclopentenylcytosine), IMP dehydrogenase, ribonucleotide reductase (a target of, e.g., carboxyl-containing N-alkyldipeptides as described in U.S. Pat. No. 5,110,799 (Tolman et al., Merck)), thymidine kinase (a target of, e.g., 1-[2-(hydroxymethyl)cycloalkylmethyl]-5-substituted-uracils and -guanines as described in, e.g., U.S. Pat. Nos. 4,863,927 and 4,782,062 (Tolman et al.; Merck)) as well as other enzymes. It will be apparent to one of ordinary skill in the art that there are additional viral proteins, both characterized and as yet to be discovered, that can serve as target for antiviral agents.
 iv) Other Agents and Modes of Antiviral Action.
 Kutapressin is a liver derivative available from Schwarz Parma of Milwaukee, Wis. in an injectable form of 25 mg/ml. The recommended dosage for herpesviruses is from 200 to 25 mg/ml per day for an average adult of 150 pounds.
 Poly(I) Poly(C12U), an accepted antiviral drug known as Ampligen from HEM Pharmaceuticals of Rockville, Md. has been shown to inhibit herpesviruses and is another antiviral agent suitable for treating KS. Intravenous injection is the preferred route of administration. Dosages from about 100 to 600 mg/m2 are administered two to three times weekly to adults averaging 150 pounds. It is best to administer at least 200 mg/m2 per week.
 Other antiviral agents reported to show activity against herpes viruses (e.g., varicella zoster and herpes simplex) and will be useful for the treatment of herpesvirus-induced KS include mappicine ketone (SmithKline Beecham); Compounds A,79296 and A,73209 (Abbott) for varicella zoster, and Compound 882C87 (Burroughs Wellcome) [see, The Pink Sheet 55(20) May 17, 1993].
 Interferon is known inhibit replication of herpes viruses. See , supra. Interferon has known toxicity problems and it is expected that second generation derivatives will soon be available that will retain interferon's antiviral properties but have reduced side affects.
 It is also contemplated that herpes virus-induced KS may be treated by administering a herpesvirus reactivating agent to induce reactivation of the latent virus. Preferably the reactivation is combined with simultaneous or sequential administration of an anti-herpesvirus agent. Controlled reactivation over a short period of time or reactivation in the presence of an antiviral agent is believed to minimize the adverse effects of certain herpesvirus infections (e.g., as discussed in PCT Application WO 93/04683). Reactivating agents include agents such as estrogen, phorbol esters, forskolin and β-adrenergic blocking agents.
 Agents useful for treatment of herpesvirus infections and for treatment of herpesvirus-induced KS are described in numerous U.S. patents. For example, ganciclovir is an example of a antiviral guanine acyclic nucleotide of the type described in U.S. Pat. Nos. 4,355,032 and 4,603,219.
 Acyclovir is an example of a class of antiviral purine derivatives, including 9-(2-hydroxyethylmethyl)adenine, of the type described in U.S. Pat. Nos. 4,287,188, 4,294,831 and 4,199,574.
 Brivudin is an example of an antiviral deoxyuridine derivative of the type described in U.S. Pat. No. 4,424,211.
 Vidarabine is an example of an antiviral purine nucleoside of the type described in British Pat. 1,159,290.
 Brovavir is an example of an antiviral deoxyuridine derivative of the type described in U.S. Pat. Nos. 4,542,210 and 4,386,076.
 BHCG is an example of an antiviral carbocyclic nucleoside analogue of the type described in U.S. Pat. Nos. 5,153,352, 5,034,394 and 5,126,345.
 HPMPC is an example of an antiviral phosphonyl methoxyalkyl derivative with of the type described in U.S. Pat. No. 5,142,051.
 CDG (Carbocyclic 2'-deoxyguanosine) is an example of an antiviral carbocyclic nucleoside analogue of the type described in U.S. Pat. Nos. 4,543,255, 4,855,466, and 4,894,458.
 Foscarnet is described in U.S. Pat. No. 4,339,445.
 Trifluridine and its corresponding ribonucleoside is described in U.S. Pat. No. 3,201,387.
 U.S. Pat. No. 5,321,030 (Kaddurah-Daouk et al.; Amira) describes the use of creatine analogs as antiherpes viral agents. U.S. Pat. No. 5,306,722 (Kim et al.; Bristol-Meyers Squibb) describes thymidine kinase inhibitors useful for treating HSV infections and for inhibiting herpes thymidine kinase. Other antiherpesvirus compositions are described in U.S. Pat. Nos. 5,286,649 and 5,098,708 (Konishi et al., Bristol-Meyers Squibb) and 5,175,165 (Blumenkopf et al.; Burroughs Wellcome). U.S. Pat. No. 4,880,820 (Ashton et al.; Merck) describes the antiherpes virus agent (5)-9-(2,3-dihydroxy-1-propoxymethyl)guanine.
 U.S. Pat. No. 4,708,935 (Suhadolnik et al.; Research Corporation) describes a 3'-deoxyadenosine compound effective in inhibiting HSV and EBV. U.S. Pat. No. 4,386,076 (Machida et al.; Yamasa Shoyu Kabushiki Kaisha) describes use of (E)-5-(2-halogenovinyl)-arabinofuranosyluracil as an antiherpesvirus agent. U.S. Pat. No. 4,340,599 (Lieb at al.; Bayer Aktiengesellschaft) describes phosphonohydroxyacetic acid derivatives useful as antiherpes agents. U.S. Pat. Nos. 4,093,715 and 4,093,716 (Lin et al. Research Corporation) describe 5'-amino-5'-deoxythymidine and 5-iodo-5'-amino-2',5'-dideoxycytidine as potent inhibitors of herpes simplex virus. U.S. Pat. No. 4,069,382 (Baker at al.; Parke, Davis & Company) describes 9-(5-O-Acyl-beta-D-arabinofuranosyladenine compounds useful as antiviral agents. U.S. Pat. No. 3,927,216 (Witkowski et al.) describes the use of 1,2,4-triazole-3-carboxamide and 1,2,4-triazole-3-thiocarboxamide for inhibiting herpes virus infections. U.S. Pat. No. 5,179,093 (Afonso et al., Schering) describes quinoline-2,4-dione derivatives active against herpes simplex virus 1 and 2, cytomegalovirus and Epstein Barr virus.
 v) Inhibitory Nucleic Acid Therapeutics
 Also contemplated here are inhibitory nucleic acid therapeutics which can inhibit the activity of herpesviruses in patients with KS. Inhibitory nucleic acids may be single-stranded nucleic acids, which can specifically bind to a complementary nucleic acid sequence. By binding to the appropriate target sequence, an RNA-RNA, a DNA-DNA, or RNA-DNA duplex or triplex is formed. These nucleic acids are often termed "antisense" because they are usually complementary to the sense or coding strand of the gene, although recently approaches for use of "sense" nucleic acids have also been developed. The term "inhibitory nucleic acids" as used herein, refers to both "sense" and "antisense" nucleic acids.
 By binding to the target nucleic acid, the inhibitory nucleic acid can inhibit the function of the target nucleic acid. This could, for example, be a result of blocking DNA transcription, processing or poly(A) addition to mRNA, DNA replication, translation, or promoting inhibitory mechanisms of the cells, such as promoting RNA degradation. Inhibitory nucleic acid methods therefore encompass a number of different approaches to altering expression of herpesvirus genes. These different types of inhibitory nucleic acid technology are described in Helene, C. and Toulme, J. , which is hereby incorporated by reference and is referred to hereinafter as "Helene and Toulme."
 In brief, inhibitory nucleic acid therapy approaches can be classified into those that target DNA sequences, those that target RNA sequences (including pre-mRNA and mRNA), those that target proteins (sense strand approaches), and those that cause cleavage or chemical modification of the target nucleic acids.
 Approaches targeting DNA fall into several categories. Nucleic acids can be designed to bind to the major groove of the duplex DNA to form a triple helical or "triplex" structure. Alternatively, inhibitory nucleic acids are designed to bind to regions of single stranded DNA resulting from the opening of the duplex DNA during replication or transcription. See Helene and Toulme.
 More commonly, inhibitory nucleic acids are designed to bind to mRNA or mRNA precursors. Inhibitory nucleic acids are used to prevent maturation of pre-mRNA. Inhibitory nucleic acids may be designed to interfere with RNA processing, splicing or translation.
 The inhibitory nucleic acids can be targeted to mRNA. In this approach, the inhibitory nucleic acids are designed to specifically block translation of the encoded protein. Using this approach, the inhibitory nucleic acid can be used to selectively suppress certain cellular functions by inhibition of translation of mRNA encoding critical proteins. For example, an inhibitory nucleic acid complementary to regions of c-myc mRNA inhibits c-myc protein expression in a human promyelocytic leukemia cell line, HL60, which overexpresses the c-myc protooncogene. See Wickstrom E. L., et al.  and Harel-Hellan, A., et al. [31A]. As described in Helene and Toulme, inhibitory nucleic acids targeting mRNA have been shown to work by several different mechanisms to inhibit translation of the encoded protein(s).
 The inhibitory nucleic acids introduced into the cell can also encompass the "sense" strand of the gene or mRNA to trap or compete for the enzymes or binding proteins involved in mRNA translation. See Helene and Toulme.
 Lastly, the inhibitory nucleic acids can be used to induce chemical inactivation or cleavage of the target genes or mRNA. Chemical inactivation can occur by the induction of crosslinks between the inhibitory nucleic acid and the target nucleic acid within the cell. Other chemical modifications of the target nucleic acids induced by appropriately derivatized inhibitory nucleic acids may also be used.
 Cleavage, and therefore inactivation, of the target nucleic acids may be effected by attaching a substituent to the inhibitory nucleic acid which can be activated to induce cleavage reactions. The substituent can be one that affects either chemical, or enzymatic cleavage. Alternatively, cleavage can be induced by the use of ribozymes or catalytic RNA. In this approach, the inhibitory nucleic acids would comprise either naturally occurring RNA (ribozymes) or synthetic nucleic acids with catalytic activity.
 The targeting of inhibitory nucleic acids to specific cells of the immune system by conjugation with targeting moieties binding receptors on the surface of these cells can be used for all of the above forms of inhibitory nucleic acid therapy. This invention encompasses all of the forms of inhibitory nucleic acid therapy as described above and as described in Helene and Toulme.
 This invention relates to the targeting of inhibitory nucleic acids to sequences the human herpesvirus of the invention for use in treating KS. An example of an antiherpes virus inhibitory nucleic acid is ISIS 2922 (ISIS Pharmaceuticals) which has activity against CMV [see, Biotechnology News 14(14) p. 5].
 A problem associated with inhibitory nucleic acid therapy is the effective delivery of the inhibitory nucleic acid to the target cell in vivo and the subsequent internalization of the inhibitory nucleic acid by that cell. This can be accomplished by linking the inhibitory nucleic acid to a targeting moiety to form a conjugate that binds to a specific receptor on the surface of the target infected cell, and which is internalized after binding.
 iii) Administration
 The subjects to be treated or whose tissue may be used herein may be a mammal, or more specifically a human, horse, pig, rabbit, dog, monkey, or rodent. In the preferred embodiment the subject is a human.
 The compositions are administered in a manner compatible with the dosage formulation, and in a therapeutically effective amount. Precise amounts of active ingredient required to be administered depend on the judgment of the practitioner and are peculiar to each subject.
 Suitable regimes for initial administration and booster shots are also variable, but are typified by an initial administration followed by repeated doses at one or more hour intervals by a subsequent injection or other administration.
 As used herein administration means a method of administering to a subject. Such methods are well known to those skilled in the art and include, but are not limited to, administration topically, parenterally, orally, intravenously, intramuscularly, subcutaneously or by aerosol. Administration of the agent may be effected continuously or intermittently such that the therapeutic agent in the patient is effective to treat a subject with Kaposi's sarcoma or a subject infected with a DNA virus associated with Kaposi's sarcoma.
 The antiviral compositions for treating herpesvirus-induced KS are preferably administered to human patients via oral, intravenous or parenteral administrations and other systemic forms. Those of skill in the art will understand appropriate administration protocol for the individual compositions to be employed by the physician.
 The pharmaceutical formulations or compositions of this invention may be in the dosage form of solid, semi-solid, or liquid such as, e.g., suspensions, aerosols or the like. Preferably the compositions are administered in unit dosage forms suitable for single administration of precise dosage amounts. The compositions may also include, depending on the formulation desired, pharmaceutically-acceptable, non-toxic carriers or diluents, which are defined as vehicles commonly used to formulate pharmaceutical compositions for animal or human administration. The diluent is selected so as not to affect the biological activity of the combination. Examples of such diluents are distilled water, physiological saline, Ringer's solution, dextrose solution, and Hank's solution. In addition, the pharmaceutical composition or formulation may also include other carriers, adjuvants; or nontoxic, nontherapeutic, nonimmunogenic stabilizers and the like. Effective amounts of such diluent or carrier are those amounts which are effective to obtain a pharmaceutically acceptable formulation in terms of solubility of components, or biological activity, etc.
V. Immunological Approaches to Therapy.
 Having identified a primary causal agent of KS in humans as a novel human herpesvirus, there are immunosuppressive therapies that can modulate the immunologic dysfunction that arises from the presence of viral infected tissue. In particular, agents that block the immunological attack of the viral infected cells will ameliorate the symptoms of KS and/or reduce the disease progress. Such therapies include antibodies that specifically block the targeting of viral infected cells. Such agents include antibodies which bind to cytokines that upregulate the immune system to target viral infected cells.
 The antibody may be administered to a patient either singly or in a cocktail containing two or more antibodies, other therapeutic agents, compositions, or the like, including, but not limited to, immuno-suppressive agents, potentiators and side-effect relieving agents. Of particular interest are immuno-suppressive agents useful in suppressing allergic reactions of a host. Immunosuppressive agents of interest include prednisone, prednisolone, DECADRON (Merck, Sharp & Dohme, West Point, Pa.), cyclophosphamide, cyclosporine, 6-mercaptopurine, methotrexate, azathioprine and i.v. gamma globulin or their combination. Potentiators of interest include monensin, ammonium chloride and chloroquine. All of these agents are administered in generally accepted efficacious dose ranges such as those disclosed in the Physician Desk Reference, 41st Ed. (1987), Publisher Edward R. Barnhart, New Jersey.
 Immune globulin from persons previously infected with human herpesviruses or related viruses can be obtained using standard techniques. Appropriate titers of antibodies are known for this therapy and are readily applied to the treatment of KS. Immune globulin can be administered via parenteral injection or by intrathecal shunt. In brief, immune globulin preparations may be obtained from individual donors who are screened for antibodies to the KS-associated human herpesvirus, and plasmas from high-titered donors are pooled. Alternatively, plasmas from donors are pooled and then tested for antibodies to the human herpesvirus of the invention; high-titered pools are then selected for use in KS patients.
 Antibodies may be formulated into an injectable preparation. Parenteral formulations are known and are suitable for use in the invention, preferably for i.m. or i.v. administration. The formulations containing therapeutically effective amounts of antibodies or immunotoxins are either sterile liquid solutions, liquid suspensions or lyophilized versions and optionally contain stabilizers or excipients. Lyophilized compositions are reconstituted with suitable diluents, e.g., water for injection, saline, 0.3% glycine and the like, at a level of about from 0.01 mg/kg of host body weight to 10 mg/kg where appropriate. Typically, the pharmaceutical compositions containing the antibodies or immunotoxins will be administered in a therapeutically effective dose in a range of from about 0.01 mg/kg to about 5 mg/kg of the treated mammal. A preferred therapeutically effective dose of the pharmaceutical composition containing antibody or immunotoxin will be in a range of from about 0.01 mg/kg to about 0.5 mg/kg body weight of the treated mammal administered over several days to two weeks by daily intravenous infusion, each given over a one hour period, in a sequential patient dose-escalation regimen.
 Antibody may be administered systemically by injection i.m., subcutaneously or intraperitoneally or directly into KS lesions. The dose will be dependent upon the properties of the antibody or immunotoxin employed, e.g., its activity and biological half-life, the concentration of antibody in the formulation, the site and rate of dosage, the clinical tolerance of the patient involved, the disease afflicting the patient and the like as is well within the skill of the physician.
 The antibody of the present invention may be administered in solution. The pH of the solution should be in the range of pH 5 to 9.5, preferably pH 6.5 to 7.5. The antibody or derivatives thereof should be in a solution having a suitable pharmaceutically acceptable buffer such as phosphate, tris (hydroxymethyl) aminomethane-HCl or citrate and the like. Buffer concentrations should be in the range of 1 to 100 mM. The solution of antibody may also contain a salt, such as sodium chloride or potassium chloride in a concentration of 50 to 150 mM. An effective amount of a stabilizing agent such as an albumin, a globulin, a gelatin, a protamine or a salt of protamine may also be included and may be added to a solution containing antibody or immunotoxin or to the composition from which the solution is prepared.
 Systemic administration of antibody is made daily, generally by intramuscular injection, although intravascular infusion is acceptable. Administration may also be intranasal or by other nonparenteral routes. Antibody or immunotoxin may also be administered via microspheres, liposomes or other microparticulate delivery systems placed in certain tissues including blood.
 In therapeutic applications, the dosages of compounds used in accordance with the invention vary depending on the class of compound and the condition being treated. The age, weight, and clinical condition of the recipient patient; and the experience and judgment of the clinician or practitioner administering the therapy are among the factors affecting the selected dosage. For example, the dosage of an immunoglobulin can range from about 0.1 milligram per kilogram of body weight per day to about 10 mg/kg per day for polyclonal antibodies and about 5% to about 20% of that amount for monoclonal antibodies. In such a case, the immunoglobulin can be administered once daily as an intravenous infusion. Preferably, the dosage is repeated daily until either a therapeutic result is achieved or until side effects warrant discontinuation of therapy. Generally, the dose should be sufficient to treat or ameliorate symptoms or signs of KS without producing unacceptable toxicity to the patient.
 An effective amount of the compound is that which provides either subjective relief of a symptom(s) or an objectively identifiable improvement as noted by the clinician or other qualified observer. The dosing range varies with the compound used, the route of administration and the potency of the particular compound.
VI. Vaccines and Prophylaxis for KS
 This invention provides a method of vaccinating a subject against Kaposi's sarcoma, comprising administering to the subject an effective amount of the peptide or polypeptide encoded by the isolated DNA molecule, and a suitable acceptable carrier, thereby vaccinating the subject. In one embodiment naked DNA is administering to the subject in an effective amount to vaccinate a subject against Kaposi's sarcoma.
 This invention provides a method of immunizing a subject against a disease caused by the DNA herpesvirus associated with Kaposi's sarcoma which comprises administering to the subject an effective immunizing dose of the isolated herpesvirus vaccine.
 A. Vaccines
 The invention also provides substances suitable for use as vaccines for the prevention of KS and methods for administering them. The vaccines are directed against the human herpesvirus of the invention, and most preferably comprise antigen obtained from the KS-associated human herpesvirus.
 Vaccines can be made recombinantly. Typically, a vaccine will include from about 1 to about 50 micrograms of antigen or antigenic protein or peptide. More preferably, the amount of protein is from about 15 to about 45 micrograms. Typically, the vaccine is formulated so that a dose includes about 0.5 milliliters. The vaccine may be administered by any route known in the art. Preferably, the route is parenteral. More preferably, it is subcutaneous or intramuscular.
 There are a number of strategies for amplifying an antigen's effectiveness, particularly as related to the art of vaccines. For example, cyclization or circularization of a peptide can increase the peptide's antigenic and immunogenic potency. See U.S. Pat. No. 5,001,049 which is incorporated by reference herein. More conventionally, an antigen can be conjugated to a suitable carrier, usually a protein molecule. This procedure has several facets. It can allow multiple copies of an antigen, such as a peptide, to be conjugated to a single larger carrier molecule. Additionally, the carrier may possess properties which facilitate transport, binding, absorption or transfer of the antigen.
 For parenteral administration, such as subcutaneous injection, examples of suitable carriers are the tetanus toxoid, the diphtheria toxoid, serum albumin and lamprey, or keyhole limpet, hemocyanin because they provide the resultant conjugate with minimum genetic restriction. Conjugates including these universal carriers can function as T cell clone activators in individuals having very different gene sets.
 The conjugation between a peptide and a carrier can be accomplished using one of the methods known in the art. Specifically, the conjugation can use bifunctional cross-linkers as binding agents as detailed, for example, by Means and Feeney, "A recent review of protein modification techniques," Bioconjugate Chem. 1:2-12 (1990).
 Vaccines against a number of the Herpesviruses have been successfully developed. Vaccines against Varicella-Zoster Virus using a live attenuated Oka strain is effective in preventing herpes zoster in the elderly, and in preventing chickenpox in both immunocompromised and normal children (Hardy, I., et al. ; Hardy, I. et al. ; Levin, M. J. et al. ; Gershon, A. A. . Vaccines against Herpes simplex Types 1 and 2 are also commercially available with some success in protection against primary disease, but have been less successful in preventing the establishment of latent infection in sensory ganglia (Roizman, B. ; Skinner, G. R. et al. ).
 Vaccines against the human herpesvirus can be made by isolating extracellular viral particles from infected cell cultures, inactivating the virus with formaldehyde followed by ultracentrifugation to concentrate the viral particles and remove the formaldehyde, and immunizing individuals with 2 or 3 doses containing 1×109 virus particles (Skinner, G. R. et al. ). Alternatively, envelope glycoproteins can be expressed in E. coli or transfected into stable mammalian cell lines, the proteins can be purified and used for vaccination (Lasky, L. A. ). MHC-binding peptides from cells infected with the human herpesvirus can be identified for vaccine candidates per the methodology of , supra.
 The antigen may be combined or mixed with various solutions and other compounds as is known in the art. For example, it may be administered in water, saline or buffered vehicles with or without various adjuvants or immunodiluting agents. Examples of such adjuvants or agents include aluminum hydroxide, aluminum phosphate, aluminum potassium sulfate (alum), beryllium sulfate, silica, kaolin, carbon, water-in-oil emulsions, oil-in-water emulsions, muramyl dipeptide, bacterial endotoxin, lipid X, Corynebacterium parvum (Propionibacterium acnes), Bordetella pertussis, polyribonucleotides, sodium alginate, lanolin, lysolecithin, vitamin A, saponin, liposomes, levamisole, DEAE-dextran, blocked copolymers or other synthetic adjuvants. Such adjuvants are available commercially from various sources, for example, Merck Adjuvant 65 (Merck and Company, Inc., Rahway, N.J.) or Freund's Incomplete Adjuvant and Complete Adjuvant (Difco Laboratories, Detroit, Mich.). Other suitable adjuvants are Amphigen (oil-in-water), Alhydrogel (aluminum hydroxide), or a mixture of Amphigen and Alhydrogel. Only aluminum is approved for human use.
 The proportion of antigen and adjuvant can be varied over a broad range so long as both are present in effective amounts. For example, aluminum hydroxide can be present in an amount of about 0.5% of the vaccine mixture (Al2O, basis). On a per-dose basis, the amount of the antigen can range from about 0.1 μg to about 100 μg protein per patient. A preferable range is from about 1 μg to about 50 μg per dose. A more preferred range is about 15 μg to about 45 μg. A suitable dose size is about 0.5 ml. Accordingly, a dose for intramuscular injection, for example, would comprise 0.5 ml containing 45 μg of antigen in admixture with 0.5% aluminum hydroxide. After formulation, the vaccine may be incorporated into a sterile container which is then sealed and stored at a low temperature, for example 4° C., or it may be freeze-dried. Lyophilization permits long-term storage in a stabilized form.
 The vaccines may be administered by any conventional method for the administration of vaccines including oral and parenteral (e.g., subcutaneous or intramuscular) injection. Intramuscular administration is preferred. The treatment may consist of a single dose of vaccine or a plurality of doses over a period of time. It is preferred that the dose be given to a human patient within the first 8 months of life. The antigen of the invention can be combined with appropriate doses of compounds including influenza antigens, such as influenza type A antigens. Also, the antigen could be a component of a recombinant vaccine which could be adaptable for oral administration.
 Vaccines of the invention may be combined with other vaccines for other diseases to produce multivalent vaccines. A pharmaceutically effective amount of the antigen can be employed with a pharmaceutically acceptable carrier such as a protein or diluent useful for the vaccination of mammals, particularly humans.
 Other vaccines may be prepared according to methods well-known to those skilled in the art.
 Those of skill will readily recognize that it is only necessary to expose a mammal to appropriate epitopes in order to elicit effective immunoprotection. The epitopes are typically segments of amino acids which are a small portion of the whole protein. Using recombinant genetics, it is routine to alter a natural protein's primary structure to create derivatives embracing epitopes that are identical to or substantially the same as (immunologically equivalent to) the naturally occurring epitopes. Such derivatives may include peptide fragments, amino acid substitutions, amino acid deletions and amino acid additions of the amino acid sequence for the viral proteins from the human herpesvirus. For example, it is known in the protein art that certain amino acid residues can be substituted with amino acids of similar size and polarity without an undue effect upon the biological activity of the protein. The human herpesvirus proteins have significant tertiary structure and the epitopes are usually conformational. Thus, modifications should generally preserve conformation to produce a protective immune response.
 B. Antibody Prophylaxis
 Therapeutic, intravenous, polyclonal or monoclonal antibodies can been used as a mode of passive immunotherapy of herpesviral diseases including perinatal varicella and CMV. Immune globulin from persons previously infected with the human herpesvirus and bearing a suitably high titer of antibodies against the virus can be given in combination with antiviral agents (e.g. ganciclovir), or in combination with other modes of immunotherapy that are currently being evaluated for the treatment of KS, which are targeted to modulating the immune response (i.e. treatment with copolymer-1, antiidiotypic monoclonal antibodies, T cell "vaccination"). Antibodies to human herpesvirus can be administered to the patient as described herein. Antibodies specific for an epitope expressed on cells infected with the human herpesvirus are preferred and can be obtained as described above.
 A polypeptide, analog or active fragment can be formulated into the therapeutic composition as neutralized pharmaceutically acceptable salt forms. Pharmaceutically acceptable salts include the acid addition salts (formed with the free amino groups of the polypeptide or antibody molecule) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed from the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, 2-ethylamino ethanol, histidine, procaine, and the like.
 C. Monitoring Therapeutic Efficacy
 This invention provides a method for monitoring the therapeutic efficacy of treatment for Kaposi's sarcoma, which comprises determining in a first sample from a subject with Kaposi's sarcoma the presence of the isolated DNA molecule, administering to the subject a therapeutic amount of an agent such that the agent is contacted to the cell in a sample, determining after a suitable period of time the amount of the isolated DNA molecule in the second sample from the treated subject, and comparing the amount of isolated DNA molecule determined in the first sample with the amount determined in the second sample, a difference indicating the effectiveness of the agent, thereby monitoring the therapeutic efficacy of treatment for Kaposi's sarcoma. As defined herein "amount" is viral load or copy number. Methods of determining viral load or copy number are known to those skilled in the art.
VII. Screening Assays for Pharmaceutical Agents of Interest in Alleviating the Symptoms of KS.
 Since an agent involved in the causation or progression of KS has been identified and described here, assays directed to identifying potential pharmaceutical agents that inhibit the biological activity of the agent are possible. KS drug screening assays which determine whether or not a drug has activity against the virus described herein are contemplated in this invention. Such assays comprise incubating a compound to be evaluated for use in KS treatment with cells which express the KS associated human herpesvirus proteins or peptides and determining therefrom the effect of the compound on the activity of such agent. In vitro assays in which the virus is maintained in suitable cell culture are preferred, though in vivo animal models would also be effective.
 Compounds with activity against the agent of interest or peptides from such agent can be screened in in vitro as well as in vivo assay systems. In vitro assays include infecting peripheral blood leukocytes or susceptible T cell lines such as MT-4 with the agent of interest in the presence of varying concentrations of compounds targeted against viral replication, including nucleoside analogs, chain terminators, antisense oligonucleotides and random polypeptides (Asada, H. et al. ; Kikuta et al.,  both incorporated by reference herein). Infected cultures and their supernatants can be assayed for the total amount of virus including the presence of the viral genome by quantitative PCR, by dot blot assays, or by using immunologic methods. For example, a culture of susceptible cells could be infected with the human herpesvirus in the presence of various concentrations of drug, fixed on slides after a period of days, and examined for viral antigen by indirect immunofluorescence with monoclonal antibodies to viral peptides (, supra. Alternatively, chemically adhered MT-4 cell monolayers can be used for an infectious agent assay using indirect immunofluorescent antibody staining to search for focus reduction (Higashi, K. et al., , incorporated by reference herein).
 As an alternative to whole cell in vitro assays, purified enzymes isolated from the human herpesvirus can be used as targets for rational drug design to determine the effect of the potential drug on enzyme activity, such as thymidine phosphotransferase or DNA polymerase. The genes for these two enzymes are provided herein. A measure of enzyme activity indicates effect on the agent itself.
 Drug screens using herpes viral products are known and have been previously described in EP 0514830 (herpes proteases) and WO 94/04920 (UL13 gene product).
 This invention provides an assay for screening anti-KS chemotherapeutics. Infected cells can be incubated in the presence of a chemical agent that is a potential chemotherapeutic against KS (e.g. acyclo-guanosine). The level of virus in the cells is then determined after several days by IFA for antigens or Southern blotting for viral genome or Northern blotting for mRNA and compared to control cells. This assay can quickly screen large numbers of chemical compounds that may be useful against KS.
 Further, this invention provides an assay system that is employed to identify drugs or other molecules capable of binding to the DNA molecule or proteins, either in the cytoplasm or in the nucleus, thereby inhibiting or potentiating transcriptional activity. Such assay would be useful in the development of drugs that would be specific against particular cellular activity, or that would potentiate such activity, in time or in level of activity.
 This invention is further illustrated in the Experimental Details section which follows. This section is set forth to aid in an understanding of the invention but is not intended to, and should not be construed to, limit in any way the invention as set forth in the claims which follow thereafter.
EXPERIMENTAL DETAILS SECTION I
Representational Difference Analysis (RDA) to Identify and Characterize Unique DNA Sequences in KS Tissue
 To search for foreign DNA sequences belonging to an infectious agent in AIDS-KS, representational difference analysis (RDA) was employed to identify and characterize unique DNA sequences in KS tissue that are either absent or present in low copy number in non-diseased tissue obtained from the same patient . This method can detect adenovirus genome added in single copy to human DNA but has not been used to identify previously uncultured infectious agents. RDA is performed by making simplified "representations" of genomes from diseased and normal tissues from the same individual through PCR amplification of short restriction fragments. The DNA representation from the diseased tissue is then ligated to a priming sequence and hybridized to an excess of unligated, normal tissue DNA representation. Only unique sequences found in the diseased tissue have priming sequences on both DNA strands and are preferentially amplified during subsequent rounds of PCR amplification. This process can be repeated using different ligated priming sequences to enrich the sample for unique DNA sequences that are only found in the tissue of interest.
 DNA (10 μg) extracted from both the KS lesion and unaffected tissue were separately digested to completion with Bam HI (20 units/μg) at 37° C. for 2 hours and 2 μg of digestion fragments were ligated to NBam12 and NBam24 priming sequences [primer sequences described in 58]. Thirty cycles of PCR amplification were performed to amplify "representations" of both genomes. After construction of the genomic representations, KS tester amplicons between 150 and 1500 bp were isolated from an agarose gel and NBam priming sequences were removed by digestion with NBam HI. To search for unique DNA sequences not found in non-KS driver DNA, a second set of priming sequences (JBam12 and JBam24) was ligated onto only the KS tester DNA amplicons (FIG. 1, lane 1). 0.2 μg of ligated KS lesion amplicons were hybridized to 20 μg of unligated, normal tissue representational amplicons. An aliquot of the hybridization product was then subjected to 10 cycles of PCR amplification using JBam24, followed by mung bean nuclease digestion. An aliquot of the mung bean-treated difference product was then subjected to 15 more cycles of PCR with the JBam24 primer (FIG. 1, lane 2). Amplification products were redigested with Bam HI and 200 ng of the digested product was ligated to RBam12 and RBam24 primer sets for a second round of hybridization and PCR amplification (FIG. 1, lane 3). This enrichment procedure was repeated a third time using the JBam primer set (FIG. 1, lane 4). Both the original driver and the tester DNA samples (Table 2, Patient A) were subsequently found to contain the AIDS-KS specific sequences KS330Bam and KS631Bam (previously identified as KS627Bam) indicating that RDA can be successfully employed when the target sequences are present in unequal copy number in both tissues.
 The initial round of DNA amplification-hybridization from KS and normal tissue resulted in a diffuse banding pattern (FIG. 1, lane 2), but four bands at approximately 380, 450, 540 and 680 bp were identifiable after the second amplification-hybridization (FIG. 1, lane 3). These bands became discrete after a third round of amplification-hybridization (FIG. 1, lane 4). Control RDA, performed by hybridizing DNA extracted from AIDS-KS tissue against itself, produced a single band at approximately 540 bp (FIG. 1, lane 5). The four KS-associated bands (designated KS330Bam, KS390Bam, KS480Bam, KS627Bam after digestion of the two flanking 28 bp ligated priming sequences with Bam HI) were gel purified and cloned by insertion into the pCRII vector. PCR products were cloned in the pCRII vector using the TA cloning system (Invitrogen Corporation, San Diego, Calif.).
Determination of the Specificity of AIDS-KS Unique Sequences
 To determine the specificity of these sequences for AIDS-KS, random-primed 32P-labeled inserts were hybridized to Southern blots of DNA extracted from cryopreserved tissues obtained from patients with and without AIDS. All AIDS-KS specimens were examined microscopically for morphologic confirmation of KS and immunohistochemically for Factor VIII, Ulex europaeus and CD34 antigen expression. One of the AIDS-KS specimens was apparently mislabeled since KS tissue was not detected on microscopic examination but was included in the KS specimen group for purposes of statistical analysis. Control tissues used for comparison to the KS lesions included 56 lymphomas from patients with and without AIDS, 19 hyperplastic lymph nodes from patients with and without AIDS, 5 vascular tumors from nonAIDS patients and 13 tissues infected with opportunistic infections that commonly occur in AIDS patients. Control DNA was also extracted from a consecutive series of 49 surgical biopsy specimens from patients without AIDS. Additional clinical and demographic information on the specimens was not collected to preserve patient confidentiality.
 The tissues, listed in Table 1, were collected from diagnostic biopsies and autopsies between 1983 and 1993 and stored at -70° C. Each tissue sample was from a different patient, except as noted in Table 1. Most of the 27 KS specimens were from lymph nodes dissected under surgical conditions which diminishes possible contamination with normal skin flora. All specimens were digested with Bam HI prior to hybridization.
 KS390Bam and KS480Bam hybridized nonspecifically to both KS and non-KS tissues and were not further characterized. 20 of 27 (74%) AIDS-KS DNAs hybridized with variable intensity to both KS330Bam and KS627Bam, and one additional KS specimen hybridized only to KS627Bam by Southern blotting (FIG. 2 and Table 1). In contrast to AIDS-KS lesions, only 6 of 39 (15%) non-KS tissues from patients with AIDS hybridized to the KS330Bam and KS627Bam inserts (Table 1).
 Specific hybridization did not occur with lymphoma or lymph node DNA from 36 persons without AIDS or with control DNA from 49 tissue biopsy specimens obtained from a consecutive series of patients. DNA extracted from several vascular tumors, including a hemangiopericytoma, two angiosarcomas and a lymphangioma, were also negative by Southern blot hybridization. DNA extracted from tissues with opportunistic infections common to AIDS patients, including 7 acid-fast bacillus (undetermined species), cytomegalovirus, 1 cat-scratch bacillus, 2 cryptococcus and 1 toxoplasmosis infected tissues, were negative by Southern blot hybridization to KS330Bam and KS627Bam (Table 1).
TABLE-US-00003 TABLE 1 Southern blot hybridization for KS330Bam and KS627Bam and PCR amplification for KS330234 in human tissues from individual patients. KS330Bam KS627Bam KS330234 Southern hybrid- Southern hybrid- PCR Tissue n ization n(%) ization n(%) positive AIDS-KS 27* 20 (74) 21 (78) 25 (93) AIDS 27† 3 (11) 3 (11) 3 (11) lymphomas AIDS 12 3 (25) 3 (25) 3 (25) lymph nodes Non-AIDS 29 0 (0) 0 (0) 0 (0) Lymphomas Non-AIDS 7 0 (0) 0 (0) 0 (0) lymph nodes Vascular 4§ 0 (0) 0 (0) 0 (0) tumors Opportu- 13π 0 (0) 0 (0) 0 (0) nistic infections Consecutive 49 ** 0 (0) 0 (0) 0 (0) surgical biopsies Legend to Table 1: *Includes one AIDS-KS specimen unamplifiable for p53 exon 6 and one tissue which on microscopic examination did not have any detectable KS tissue present. Both of these samples were negative by Southern blot hybridization to KS330Bam and KS627Bam and by PCR amplification for the KS330234 amplicon. †Includes 7 small non-cleaved cell lymphomas, 20 diffuse large cell and immunoblastic lymphomas. Three of the lymphomas with immunoblastic morphology were positive for KS330Bam and KS627Bam. .dagger-dbl.Includes 13 anaplastic large cell lymphomas, 4 diffuse large cell lymphomas, 4 small lymphocytic lymphomas/chronic lymphocytic leukemias, 3 hairy cell leukemias, 2 monocytoid B-cell lymphomas, 1 follicular small cleaved cell lymphoma, 1 Burkitt's lymphoma, 1 plasmacytoma. §Includes 2 angiosarcomas, 1 hemangiopericytoma and 1 lymphangioma. πIncludes 2 cryptococcus, 1 toxoplasmosis, 1 cat-scratch bacillus, 1 cytomegalovirus, 1 Epstein-Barr virus, and 7 acid-fast bacillus infected tissues. In addition, pure cultures of Mycobacterium avium-complex were negative by Southern hybridization and PCR, and pure cultures of Mycoplasma penetrans were negative by PCR. Tissues included skin, appendix, kidney, prostate, hernia sac, lung, fibrous tissue, gallbladder, colon, foreskin, thyroid, small bowel, adenoid, vein, axillary tissue, lipoma, heart, mouth, hemorrhoid, pseudoaneurysm and fistula track. Tissues were collected from a consecutive series of biopsies on patients without AIDS but with unknown HIV serostatus. **Apparent nonspecific hybridization at approximately 20 Kb occurred in 4 consecutive surgical biopsy DNA samples: one colon and one hernia sac DNA sample hybridized to KS330Bam alone, another hernia sac DNA sample hybridized to KS627Bam alone and one appendix DNA sample hybridized to both KS330Bam and KS627Bam. These samples did not hybridize in the 330-630 bp range expected for these sequences and were PCR negative for KS330234.
 In addition, DNA from Epstein-Barr virus-infected peripheral blood lymphocytes and pure cultures of Mycobacterium avium-complex were also negative by Southern hybridization. Overall, 20 of 27 (74%) AIDS-KS specimens hybridized to KS330Bam and 21 of 27 (78%) AIDS-KS specimens hybridized to KS627Bam, compared to only 6 of 142 (4%) non-KS human DNA control specimens (χ2=85.02, p<10-7 and χ2=92.4, p<10-7 respectively).
 The sequence copy number in the AIDS-KS tissues was estimated by simultaneous hybridization with KS330Bam and a 440 bp probe for the constant region of the T cell receptor β gene . Samples in lanes 5 and 6 of FIGS. 2A-2B showed similar intensities for the two probes indicating an average copy number of approximately two KS330Bam sequences per cell, while remaining tissues had weaker hybridization signals for the KS330Bam probe.
Characterization of KS330Bam and KS627Bam
 To further characterize KS330Bam and KS627Bam, six clones for each insert were sequenced. The Sequenase version 2.0 (United States Biochemical, Cleveland, Ohio) system was used and sequencing was performed according to manufacturer's instructions. Nucleotides sequences were confirmed with an Applied Biosystems 373A Sequencer in the DNA Sequencing Facilities at Columbia University.
 KS330Bam is a 330 bp sequence with 51% G:C content (FIG. 3B) and KS627Bam is a 627 bp sequence with a 63% G:C content (FIG. 3C). KS330Bam has 54% nucleotide identity to the BDLF1 open reading frame (ORF) of Epstein-Barr virus (EBV). Further analysis revealed that both KS330Bam and KS627Bam code for amino acid sequences with homology to polypeptides of viral origin. SwissProt and PIR protein databases were searched for homologous ORF using BLASTX .
 KS330Bam is 511 identical by amino acid homology to a portion of the ORF26 open reading frame encoding the capsid protein VP23 (NCBI g.i. 60348, bp 46024-46935) of herpesvirus saimiri , a gammaherpesvirus which causes fulminant lymphoma in New world monkeys. This fragment also has a 39% identical amino acid sequence to the theoretical protein encoded by the homologous open reading frame BDLF1 in EBV (NCBI g.i. 59140, bp 132403-133307) . The amino acid sequence encoded by KS627Bam is homologous with weaker identity (31%) to the tegument protein, gp140 (ORF 29, NCBI g.i. 60396, bp108782-112681) of herpesvirus saimiri.
 Sequence data from KS330Bam was used to construct PCR primers to amplify a 234 bp fragment designated KS330234 (FIG. 3B). The conditions for PCR analyses were as follows: 94° C. for 2 min (1 cycle); 94° C. for 1 min, 58° C. for 1 min, 72° C. for 1 min (35 cycles); 72° C. extension for 5 min (1 cycle). Each PCR reaction used 0.1 μg of genomic DNA, 50 pmoles of each primer, 1 unit of Taq polymerase, 100 μM of each deoxynucleotide triphosphate, 50 mM KCl, 10 mM Tris-HCl (pH 9.0), and 0.1% Triton-X-100 in a final volume of 25 μl. Amplifications were carried out in a Perkin-Elmer 480 Thermocycler with 1-s ramp times between steps.
 Although Southern blot hybridization detected the KS330Bam sequence in only 20 of 27 KS tissues, 25 of the 27 tissues were positive by PCR amplification for KS330234 (FIGS. 4A-4B) demonstrating that KS330Bam is present in some KS lesions at levels below the threshold for detection by Southern blot hybridization. All KS330234 PCR products hybridized to a 32P end-labelled 25 bp internal oligomer, confirming the specificity of the PCR (FIG. 4B). Of the two AIDS-KS specimens negative for KS330234, both specimens appeared to be negative for technical reasons: one had no microscopically detectable KS tissue in the frozen sample (FIGS. 4A-4B, lane 3), and the other (FIGS. 4A-4B, lane 15) was negative in the control PCR amplification for the p53 gene indicating either DNA degradation or the presence of PCR inhibitors in the sample. PCR amplification of the p53 tumor suppressor gene was used as a control for DNA quality. Sequences of p53 primers from P6-5,5'-ACAGGGCTGGTTGCCCAGGGT-3'(SEQ ID No: 44); and P6-3. 5'-AGTTGCAAACCAGACCTCAG-3'(SEQ ID NO: 45) .
 Except for the 6 control samples from AIDS patients that were also positive by Southern blot hybridization, none of the other 136 control specimens were positive by PCR for KS330234. All of these specimens were amplifiable for the p53 gene, indicating that inadequate PCR amplification was not the reason for lack of detection of KS330234 in the control tissues. Samples containing DNA from two candidate KS agents, EBV and Mycoplasma penetrans (ATCC Accession No. 55252), a pathogen commonly found in the genital tract of patients with AIDS-KS  were also negative for amplification of KS330234. In addition, several KS specimens were tested using commercial PCR primers (Stratagene, La Jolla, Calif.) specific for mycoplasmata and primers specific for the EBNA-2, EBNA-3C and EBER regions of EBV and were negative .
 Overall, DNA from 25 (93%) of 27 AIDS-KS tissues were positive by PCR compared with DNA from 6 (4%) of 142 control tissues, including 6 (15%) of 39 non-KS lymph nodes and lymphomas from AIDS patients (χ2=38.2, p<10-6), 0 of 36 lymph nodes and lymphomas from nonAIDS patients (χ2=55.2, p<10-7) and 0 of 49 consecutive biopsy specimens (χ2=67.7, p<10-7). Thus, KS330234 was found in all 25 amplifiable tissues with microscopically detectable AIDS-KS, but rarely occurred in non-KS tissues, including tissues from AIDS patients.
 Of the six control tissues from AIDS patients that were positive by both PCR and Southern hybridization, two patients had KS elsewhere, two did not develop KS and complete clinical histories for the remaining two patients were unobtainable. Three of the six positive non-KS tissues were lymph nodes with follicular hyperplasia taken from patients with AIDS. Given the high prevalence of KS among patients with AIDS, it is possible that undetected microscopic foci of KS were present in these lymph nodes. The other three positive tissue specimens were B cell immunoblastic lymphomas from AIDS patients. It is possible that the putative KS agent is also a cofactor for a subset of AIDS-associated lymphomas [16, 17, 80].
 To determine whether KS330Bam and KS627Bam are portions of a larger genome and to determine the proximity of the two sequences to each other, samples of KS DNA were digested with Pvu II restriction enzymes. Digested genomic DNA from three AIDS-KS samples were hybridized to KS330Bam and KS627Bam by Southern blotting (FIG. 5). These sequences hybridized to various sized fragments of the digested KS DNA indicating that both sequences are fragments of larger genomes. Differences in the KS330Bam hybridization pattern to Pvu II digests of the three AIDS-KS specimens indicate that polymorphisms may occur in the larger genome. Individual fragments from the digests failed to simultaneously hybridize with both KS330Bam and KS627Bam, demonstrating that these two Bam HI restriction fragments are not adjacent to one another.
 If KS330Bam and KS627Bam are heritable polymorphic DNA markers for KS, these sequences should be uniformly detected at non-KS tissue sites in patients with AIDS-KS. Alternatively, if KS330Bam and KS627Bam are sequences specific for an exogenous infectious agent, it is likely that some tissues are uninfected and lack detectable KS330Bam and KS627Bam sequences. DNA extracted from multiple uninvolved tissues from three patients with AIDS-KS were hybridized to 32P-labelled KS330Bam and KS627Bam probes as well as analyzed by PCR using the KS330234 primers (Table 2). While KS lesion DNA samples were positive for both bands, unaffected tissues were frequently negative for these sequences. KS lesions from patients A, B and C, and uninvolved skin and muscle from patient A were positive for KS330Bam and KS627Bam, but muscle and brain tissue from patient B and muscle, brain, colon, heart and hilar lymph node tissues from patient C were negative for these sequences. Uninvolved stomach lining adjacent to the KS lesion in patient C was positive by PCR, but negative by Southern blotting which suggests the presence of the sequences in this tissue at levels below the detection threshold for Southern blotting.
TABLE-US-00004 TABLE 2 Differential detection of KS330Bam, KS627Bam and KS330234 sequences in KS-involved and non-involved tissues from three patients with AIDS-KS. KS330Bam KS627Bam KS330234 Patient A KS, skin + + + nl skin + + + nl muscle + + + Patient B KS, skin + + + nl muscle - - - nl brain - - - Patient C KS, stomach + + + nl stomach - - + adjacent to KS nl muscle - - - nl brain - - - nl colon - - - nl heart - - - nl hilar lymph - - - nodes
Subcloning and Sequencing of KSHV
 KS330Bam and KS627Bam are genomic fragments of a novel infectious agent associated with AIDS-KS. A genomic library from a KS lesion was made and a phage clone with a 20 kb insert containing the KS330Bam sequence was identified. The 20 kb clone digested with PvuII (which cuts in the middle of the KS330Bam sequence) produced 1.1 kb and 3 kb fragments that hybridized to KS330Bam. The 1.1 kb subcloned insert and ˜900 bp from the 3 kb subcloned insert resulting in 9404 bp of contiguous sequence was entirely sequenced. This sequence contains partial and complete open reading frames homologous to regions in gamma herpesviruses.
 The KS330Bam sequence is an internal portion of a 918 bp ORF with 55-56% nucleotide identity to the ORF26 and BDLF1 genes of HSVSA and EBV respectively (SEQ IS NO:46 and 47, respectively). The EBV and HSVSA translated amino acid sequences for these ORFs demonstrate extensive homology with the amino acid sequence encoded by the KS-associated 918 bp ORF (FIG. 6). In HSVSA, the VP23 protein is a late structural protein involved in capsid construction. Reverse transcriptase (RT)-PCR of mRNA from a KS lesion is positive for transcribed KS330Bam 15 mRNA and that indicates that this ORF is transcribed in KS lesions. Additional evidence for homology between the KS agent and herpesviruses comes from a comparison of the genomic organization of other potential ORFs on the 9404 bp sequence (FIG. 3A) The 5' terminus of the sequence is composed nucleotides having 66-67% nucleotide identity and 68-71% amino acid identity to corresponding regions of the major capsid protein (MCP) ORFs for both EBV and HSVSA. This putative MCP ORF of the KS agent lies immediately 5' to the BDLF1/ORF26 homolog which is a conserved orientation among herpesvirus subfamilies for these two genes. At the 3 end of this sequence, the reading frame has strong amino acid and nucleotide homology to HSVSA ORF 27. Thus, KS-associated DNA sequences at four loci in two separate regions with homologies to gamma herpesviral genomes have been identified.
 In addition to fragments obtained from Pvu II digest of the 21 Kb phage insert described above, fragments obtained from a BamHI/NotI digest were also subcloned into pBluescript (Stratagene, La Jolla, Calif.). The termini of these subcloned fragments were sequenced and were also found to be homologous to nucleic acid sequence EBV and HSVSA genes. These homologs have been used to develop a preliminary map of subcloned fragments (FIG. 9). Thus, sequencing has revealed that the KS agent maintains co-linear homology to gamma herpesviruses over the length of the 21 Kb phage insert.
Determination of the Phylogeny of KSHV
 Regions flanking KS330Bam were sequenced and characterized by directional walking. This was performed by the following strategy: 1) KS genomic libraries were made and screened using the KS330Bam fragment as a hybridization probe, 2) DNA inserts from phage clones positive for the KS330Bam probe were isolated and digested with suitable restriction enzyme(s), 3) the digested fragments were subcloned into pBluescript (Stratagene, La Jolla, Calif.), and 4) the subclones were sequenced. Using this strategy, the major capsid protein (MCP) ORF homolog was the first important gene locus identified. Using sequenced unique 3' and 5' end-fragments from positive phage clones as probes, and following the strategy above a KS genomic library are screened by standard methods for additional contiguous sequences.
 For sequencing purposes, restriction fragments are subcloned into phagemid pBluescript KS+, pBluescript KS-, pBS+, or pBS- (Stratagene) or into plasmid pUC18 or pUC19. Recombinant DNA was purified through CsCl density gradients or by anion-exchange chromatography (Qiagen).
 Nucleotide sequenced by standard screening methods of cloned fragments of KSHV were done by direct sequencing of double-stranded DNA using oligonucleotide primers synthesized commercially to "walk" along the fragments by the dideoxy-nucleotide chain termination method. Junctions between clones are confirmed by sequencing overlapping clones.
 Targeted homologous genes in regions flanking KS330Bam include, but are not limited to: 11-10 homolog, thymidine kinase (TK), g85, g35, gH, capsid proteins and MCP. TK is an early protein of the herpesviruses functionally linked to DNA replication and a target enzyme for anti-herpesviral nucleosides. TK phosphorylates acyclic nucleosides such as acyclovir which in turn inhibit viral DNA polymerase chain extension. Determining the sequence of this gene will aid in the prediction of chemotherapeutic agents useful against KSHV. TK is encoded by the EBV BXLF1 ORF located ˜9700 bp rightward of BDLF1 and by the HSVSA ORF 21-9200 bp rightward of the ORF 26. A subcloned fragment of KS5 was identified with strong homology to the EBV and HSVSA TK open reading frames.
 g85 is a late glycoprotein involved in membrane fusion homologous to gH in HSV1. In EBV, this protein is encoded by BLXF2 ORF located -7600 bp rightward of BDLF1, and in HSVSA it is encoded by ORF 22 located ˜7100 bp rightward of ORF26.
 g35 is a late EBV glycoprotein found in virion and plasma membrane. It is encoded by BDLF3 ORF which is 1300 bp leftward of BDLF1 in EBV. There is no BDLF3 homolog in HSVSA. A subcloned fragment has already been identified with strong homology to the EBV gp35 open reading frame.
 Major capsid protein (MCP) is a conserved 150 KDa protein which is the major component of herpesvirus capsid. Antibodies are generated against the MCP during natural infection with most herpesviruses. The terminal 1026 bp of this major capsid gene homolog in KSHV have been sequenced.
 Targeted homologous genes/loci in regions flanking KS627Bam include, but are not limited to terminal reiterated repeats, LMPI, EBERs and Ori P. Terminal reiterated sequences are present in all herpesviruses. In EBV, tandomly reiterated 0.5 Kb long terminal repeats flank the ends of the linear genome and become joined in the circular form. The terminal repeat region is immediately adjacent to BNRF1 in EBV and ORF 75 in HSVSA. Since the number of terminal repeats varies between viral strains, identification of terminal repeat regions may allow typing and clonality studies of KSHV in KS legions. Sequencing through the terminal repeat region may determine whether this virus is integrated into human genome in KS.
 LMPI is an latent protein important in the transforming effects of EBV in Burkitt's lymphoma. This gene is encoded by the EBV BNRF1 ORF located ˜2000 bp rightward of tegument protein ORF BNRF1 in the circularized genome. There is no LMP1 homolog in HSVSA.
 EBERs are the most abundant RNA in latently EBV infected cells and Ori-P is the origin of replication for latent EBV genome. This region is located between ˜4000-9000 bp leftward of the BNRF1 ORF in EBV; there are no corresponding regions in HSVSA.
 The data indicates that the KS agent is a new human herpesvirus related to gamma herpesviruses EBV and HSVSA. The results are not due to contamination or to incidental co-infection with a known herpesvirus since the sequences are distinct from all sequenced herpesviral genomes (including EBV, CMV, HHV6 and HSVSA) and are associated specifically with KS in three separate comparative studies. Furthermore, PCR testing of KS DNA with primers specific for EBV-1 and EBV-2 failed to demonstrate these viral genomes in these tissues. Although KSHV is homologous to EBV regions, the sequence does not match any other known sequence and thus provides evidence for a new viral genome, related to but distinct from known members of the herpesvirus family.
Indirect Immunofluorescence Assay (IFA)
 Virus-containing cells are coated to a microscope slide. The slides are treated with organic fixatives, dried and then incubated with patient sera. Antibodies in the sera bind to the cells, and then excess nonspecific antibodies are washed off. An antihuman immunoglobulin linked to a fluorochrome, such as fluorescein, is then incubated with the slides, and then excess fluorescent immunoglobulin is washed off. The slides are then examined under a microscope and if the cells fluoresce, then this indicates that the sera contains antibodies directed against the antigens present in the cells, such as the virus.
 An indirect immunofluorescence assay (IFA) was performed on the Body Cavity-Based Lymphoma cell line (BCBL-1), which is a naturally transformed EBV infected (nonproducing) B cell line, using 4 KS patient sera and 4 control sera (from AIDS patients without KS). Initially, both sets of sera showed similar levels of antibody binding. To remove nonspecific antibodies directed against EBV and lymphocyte antigens, sera at 1:25 dilution were pre-adsorbed using 3×106 1% paraformaldehyde-fixed Raji cells per ml of sera. BCBL1 cells were fixed with ethanol/acetone, incubated with dilutions of patient sera, washed and incubated with fluorescein-conjugated goat anti-human IgG. Indirect immunofluorescent staining was determined.
 Table 3 shows that unabsorbed case and control sera have similar end-point dilution indirect immunofluorescence assay (IFA) titers against the BCBL1 cell line. After Raji adsorption, case sera have four-fold higher IFA titers against BCBL1 cells than control sera. Results indicated that pre-adsorption against paraformaldehyde-fixed Raji cells reduces fluorescent antibody binding in control sera but do not eliminate antibody binding to KS case sera. These results indicate that subjects with KS have specific antibodies directed against the KS agent that can be detected in serological assays such as IFA, Western blot and Enzyme immunoassays (Table 3).
TABLE-US-00005 TABLE 3 Indirect immunofluorescence end-point titers for KS case and non-KS control sera against the BCBL-1 cell line Sera No. Status* Pre-adsorption Post-adsorption** 1 KS ≧1:400.sup. ≧1:400 2 KS 1:100 1:100 3 KS 1:200 1:100 4 KS ≧1:400.sup. 1:200 5 Control ≧1:400.sup. 1:50 6 Control 1:50 1:50 7 Control 1:100 1:50 8 Control 1:200 1:50 Legend Table 3: *KS = autopsy-confirmed male, AIDS patient Control = autopsy-confirmed female, AIDS patient, no KS **Adsorbed against RAJI cells treated with 1% paraformaldehyde
Immunoblotting ("Western Blot")
 Virus-containing cells or purified virus (or a portion of the virus, such as a fusion protein) is electrophoresed on a polyacrylamide gel to separate the protein antigens by molecular weight. The proteins are blotted onto a nitrocellulose or nylon membrane, then the membrane is incubated in patient sera. Antibodies directed against specific antigens are developed by incubating with a anti-human immunoglobulin attached to a reporter enzyme, such as a peroxidase. After developing the membrane, each antigen reacting against antibodies in patient sera shows up as a band on the membrane at the corresponding molecular weight region.
Enzyme Immunoassay ("EIA or ELISA")
 Virus-containing cells or purified virus (or a portion of the virus, such as a fusion protein) is coated to the bottom of a 96-well plate by various means (generally incubating in alkaline carbonate buffer). The plates are washed, then the wells are incubated with patient sera. Antibodies in the sera directed against specific antigens stick on the plate. The wells are washed again to remove nonspecific antibody, then they are incubated with a antihuman immunoglobulin attached to a reporter enzyme, such as a peroxidase. The plate is washed again to remove nonspecific antibody and then developed. Wells containing antigen that is specifically recognized by antibodies in the patients sera change color and can be detected by an ELISA plate reader (a spectrophotomer).
 All three of these methods can be made more specific by pre-incubating patient sera with uninfected cells to adsorb out cross-reacting antibodies against the cells or against other viruses that may be present in the cell line, such as EBV. Cross-reacting antibodies can potentially give a falsely positive test result (i.e. the patient is actually not infected with the virus but has a positive test result because of cross-reacting antibodies directed against cell antigens in the preparation). The importance of the infection experiments with Raji is that if Raji cells, or another well-defined cell line, can be infected, then the patient's sera can be pre-adsorbed against the uninfected parental cell line and then tested in one of the assays. The only antibodies left in the sera after pre-adsorption that bind to antigens in the preparation should be directed against the virus.
 BCBL 1, from lymphomatous tissues belonging to a rare infiltrating, anaplastic body cavity lymphoma occurring in AIDS patients has been placed in continuous cell culture and shown to be continuously infected with the KS agent. This cell line is also naturally infected with Epstein-Barr Virus (EBV). The BCBL cell line was used as an antigen substrate to detect specific KS antibodies in persons infected with the putative virus by Western-blotting. Three lymphoid B cell lines were used as controls. These included the EBV genome positive cell line P3H3, the EBV genome defective cell line Raji and the EBV genome negative cell line Bjab.
 Cells from late-log phase culture were washed 3 time with PBS by centrifugation at 500 g for 10 min. and suspended in sample buffer containing 50 mM Tris-HCl pH 6.8, 2% SDS (w/v), 15% glycerol (v/v), 5% β-mercaptoethanol (v/v) and 0.001% bromophenol (w/v) with protease inhibitor, 100 μM phenylmethylsulfonyl fluoride (PMSF). The sample was boiled at 100° C. for 5 min and centrifuged at 14,000 g for 10 min. The proteins in the supernatant was then fractionated by sodium, dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions with a separation gel of 15% and a stacking gel of 5% (3). Prestained protein standards were included: myosin, 200 kDa; β-galactosidase, 118 kDA; BSA, 78 kDa; ovalbumin, 47.1 kDa; carbonic anhydrase, 31.4 kDa; soybean trypsin inhibitor, 25.5 kDa, lysozyme, 18.8 kDa and aprotinin, 8.3 kDa (Bio-Rad). Immunoblotting experiments were performed according to the method of Towbin et al. (4). Briefly, the proteins were electrophoretically transferred to Hybon-C extra membranes (Pharmacia) at 24 V for 70 min. The membranes were then dried at 37° C. for 30 min, saturated with 5% skim milk in Tris-buffered saline, pH 7.4 (TBS) containing 50 mM Tris-HCl and 200 mM NaCl, at room temperature for 1 h. The membranes were subsequently incubated with human sera at dilution 1:200 in 1% skim milk overnight at room temperature, washed 3 times with a solution containing TBS, 0.2% Triton X-100 and 0.05% skim milk and then 2 times with TBS. The membranes were then incubated for 2 h at room temperature with alkaline phosphatase conjugated goat anti-mouse IgG+IgM+IgA (Sigma) diluted at 1:5000 in 1% skim milk. After repeating the washing, the membranes were stained with nitroblue tetranolium chloride and 5-bromo-4-chloro-3-indolylphosphate p-toluidine salt (Gibco BRL).
 Two bands of approximately 226 kDa and 234 kDa were identified to be specifically present on the Western-blot of BCBL cell lysate in 5 sera from AIDS gay man patients infected with KS. These 2 bands were absent from the lysates of P3H3, Raji and Bjab cell lysates. 5 sera from AIDS gay man patients without KS and 2 sera from AIDS woman patients without KS as well as 1 sera from nasopharyncel carcinoma patient were not able to detect these 2 bands in BCBL 1, P3H3, Raji and Bjab cell lysates. In a blinded experiment, using the 226 kDa and 234 kDa markers, 15 out of 16 sera from KS patients were correctly identified. In total, the 226 kDa and 234 kDa markers were detected in 20 out of 21 sera from KS patients.
 The antigen is enriched in the nuclei fraction of BCBL1. Enriched antigen with low background can be obtained by preparing nucleic from BCBC as the starting antigen preparation using standard, widely available protocols. For example, 500-750 ml of BCBL at 5×105 cells/ml can be pelleted at low speed. The pellet is placed in 10 mM NaCl, 10 mM This pH 7.8, 1.5 mM MgCl2 (equi volume)+1.0% NP-40 on ice for 20 min to lyse cells. The lysate is then spun at 1500 rpm for 10 min. to pellet nucleic. The pellet is used as the starting fraction for the antigen preparation for the Western blot. This will reduce cross-reactive cytoplasmic antigens.
 BCBL1 cells were co-cultivated with Raji cell lines separated by a 0.45μ tissue filter insert. Approximately, 1-2×106 BCBL1 and 2×106 Raji cells were co-cultivated for 2-20 days in supplemented RPMI alone, in 10 μg/ml 5'-bromodeoxyuridine (BUdR) and 0.6 μg/ml 5'-fluorodeoxyuridine or 20 ng/ml 12-O-tetradecanoylphorbol-13-acetate (TPA). After 2, 8, 12 or 20 days co-cultivation, Raji cells were removed, washed and placed in supplemented RPMI 1640 media. A Raji culture co-cultivated with BCBL1 in 20 ng/ml TPA for 2 days survived and has been kept in continuous suspension culture for >10 weeks. This cell line, designated RCC1 (Raji Co-Culture, No. 1) remains PCR positive for the KS330234 sequence after multiple passages. This cell line is identical to its parental Raji cell line by flow cytometry using EMA, B1, B4 and BerH2 lymphocyte-flow cytometry (approximately 2%). RCC1 periodically undergo rapid cytolysis suggestive of lytic reproduction of the agent. Thus, RCC1 is a Raji cell line newly infected with KSHV.
 The results indicate the presence of a new human virus, specifically a herpesvirus in KS lesions. The high degree of association between this agent and AIDS-KS (>90%), and the low prevalence of the agent in non-KS tissues from immunocompromised AIDS patients, indicates that this agent has a causal role in AIDS-KS [47, 68].
Isolation of KSHV
 Crude virus preparations are made from either the supernatant or low speed pelleted cell fraction of BCBL1 cultures. Approximately 650 ml or more of log phase cells should be used (>5×106 cells/ml).
 For bonding whole virion from supernatant, the cell free supernatant is spun at 10,000 rpm in a GSA rotor for 10 min to remove debris. PEG-8000 is added to 7%, dissolved and placed on ice for >2.5 hours. The PEG-supernatant is then spun at 10,000×g for 30 min. supernatant is poured off and the pellet is dried and scraped together from the centrifuge bottles. The pellet is then resuspended in a small volume (1-2 ml) of virus buffer (VB, 0.1 M NaCl, 0.01 M Tris, pH 7.5). This procedure will precipitate both naked genome and whole virion. The virion are then isolated by centrifugation at 25,000 rpm in a 10-50% sucrose gradient made with VB. One ml fractions of the gradient are then obtained by standard techniques (e.g. using a fractionator) and each fraction is then tested by dot blotting using specific hybridizing primer sequences to determine the gradient fraction containing the purified virus (preparation of the fraction maybe needed in order to detect the presence of the virus, such as standard DNA extraction).
 To obtain the episomal DNA from the virus, the pellet of cells is washed and pelleted in PBS, then lysed using hypotonic shock and/or repeated cycles of freezing and thawing in a small volume (<3 ml).
 Nuclei and other cytoplasmic debris are removed by centrifugation at 10,000 g for 10 min, filtration through a 0.45 m filter and then repeat centrifugation at 10,000 g for 10 min. This crude preparation contains viral genome and soluble cell components. The genome preparation can then be gently chloroform-phenol extracted to remove associated proteins or can be placed in neutral DNA buffer (1 M NaCl, 50 mM Tris, 10 mM EDTA, pH 7.2-7.6) with 2% sodium dodecylsulfate (SDS) and 1% sarcosyl. The genome is then banded by centrifugation through 10-30% sucrose gradient in neutral DNA buffer containing 0.15% sarcosyl at 20,000 rpm in a SW 27.1 rotor for 12 hours (for 40,000 rpm for 2-3 hours in an SW41 rotor). The band is detected as described above.
 An example of the method for isolating KSHV genome from KSHV infected cell cultures (97 and 98). Approximately 800 ml of BCBL1 cells are pelleted, washed with saline, and pelleted by low speed centrifugation. The cell pellet is lysed with an equal volume of RSB (10 mM NaCl, 10 mM Tris-HCl, 1.5 mM MgCl2, pH 7.8) with 1% NP-40 on ice for 10 minutes. The lysate is centrifuged at 900×g for 10 minutes to pellet nuclei. This step is repeated. To the supernatant is added 0.4% sodium dodecylsulfate and EDTA to a final concentration of 10 mM. The supernatant is loaded on a 10-30% sucrose gradient in 1.0 M NaCl, 1 mM EDTA, 50 mM Tris-HCl, pH 7.5. The gradients are centrifuged at 20,000 rpm on a SW 27.1 rotor for 12 hours. In FIG. 11, 0.5 ml aliquots of the gradient have been fractionated (fractions 1-62) with the 30% gradient fraction being at fraction No. 1 and the 10% gradient fraction being at fraction No. 62. Each fraction has been dot hybridized to a nitrocellulose membrane and then a 32P-labeled KSHV DNA fragment, KS631Bam has been hybridized to the membrane using standard techniques. FIG. 11 shows that the major solubilized fraction of the KSHV genome bands (i.e. is isolated) in fractions 42 through 48 of the gradient with a high concentration of the genome being present in fraction 44. A second band of solubilized KSHV DNA occurs in fractions 26 through 32.
Purification of KSHV
 DNA is extracted using standard techniques from the RCC-1 or RCC-12F5 cell line [27, 49, 66]. The DNA is tested for the presence of the KSHV by Southern blotting and PCR using the specific probes as described hereinafter. Fresh lymphoma tissue containing viable infected cells is simultaneously filtered to form a single cell suspension by standard techniques [49, 66]. The cells are separated by standard Ficoll-Plaque centrifugation and lymphocyte layer is removed. The lymphocytes are then placed at >1×106 cells/ml into standard lymphocyte tissue culture medium, such as RMP 1640 supplemented with 10% fetal calf serum. Immortalized lymphocytes containing the KSHV virus are indefinitely grown in the culture media while nonimmortilized cells die during course of prolonged cultivation.
 Further, the virus may be propagated in a new cell line by removing media supernatant containing the virus from a continuously infected cell line at a concentration of >1×106 cells/ml. The media is centrifuged at 2000×g for 10 minutes and filtered through a 0.45μ filter to remove cells. The media is applied in a 1:1 volume with cells growing at >1×106 cells/ml for 48 hours. The cells are washed and pelleted and placed in fresh culture medium, and tested after 14 days of growth.
 The herpesvirus may be isolated from the cell DNA in the following manner. An infected cell line, which can be lysed using standard methods such as hyposmotic shocking and Dounce homogenization, is first pelleted at 2000×g for 10 minutes, the supernatant is removed and centrifuged again at 10,000×g for 15 minutes to remove nuclei and organelles. The supernatant is filtered through a 0.45μ filter and centrifuged again at 100,000×g for 1 hour to pellet the virus. The virus can then be washed and centrifuged again at 100,000×g for 1 hour.
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EXPERIMENTAL DETAILS SECTION II
 Sequencing Studies: A lambda phage (KS5) from a KS lesion genomic library identified by positive hybridization with KS330Bam was digested with BamHI and Not I (Boehringer-Mannheim, Indianapolis Ind.); five fragments were gel isolated and subcloned intra Bluescript II KS (Stratagene, La Jolla Calif.). The entire sequence was determined by bidirectional sequencing at a seven fold average redundancy by primer walking and nested deletions.
 DNA sequence data were compiled and aligned using ALIGN (IBI-Kodak, Rochester N.Y.) and analyzed using the Wisconsin Sequence Analysis Package Version 8-UNIX (Genetics Computer Group, Madison Wis.) and the GRAIL Sequence Analysis, Gene Assembly and Sequence Comparison System v. 1.2 (Informatics Group, Oak Ridge Tenn.). Protein site motifs were identified using Motif (Genetics Computer Group, Madison Wis.).
 Sources of Herpesvirus Gene Sequence Comparisons: Complete genomic sequences of three gammaherpesviruses were available: Epstein-Barr virus (EBV), a herpesvirus of humans ; herpesvirus saimiri (HVS), a herpesvirus of the New World monkey Saimiri sciureus ; and equine herpesvirus 2 (EHV2 ). Additional thymidine kinase gene sequences were obtained for alcelaphine herpesvirus 1 (AHV1 ) and bovine herpesvirus 4 (BHV4 ). Sequences for the major capsid protein genes of human herpesvirus 6B and human herpesvirus 7 (HHV7) were from Mukai at al. . The sources of all other sequences used are listed previously in McGeoch and Cook  and McGeoch et al. .
 Phylogenetic Inference: Predicted amino acid sequences used for tree construction were based on previous experience with herpesviral phylogenetic analyses . Alignments of homologous sets of amino acid sequences were made with the AMPS  and Pileup  programs. Regions of alignments that showed extreme divergence with marked length heterogeneity, typically terminal sections, were excised. Generally, positions in alignments that contained inserted gaps in one or more sequences were removed before use for tree construction. Phylogenetic inference programs were from the Phylip set, version 3.5c  and from the GCG set . Trees were built with the maximum parsimony (MP), neighbor joining (NJ) methods. For the NJ method, which utilizes estimates of pairwise distances between sequences, distances were estimated as mean numbers of substitution events per site with Protdist using the PAM 250 substitution probability matrix of Schwartz & Dayhoff . Bootstrap analysis  was carried out for MP and NJ trees, with 100 sub-replicates of each alignment, and consensus trees obtained with the program Consense. In addition the program Protml was used to infer trees by the maximum likelihood (ML) method. Protml was obtained form J. Adachi, Department of Statistical Science, The Graduate University for Advanced Study, Tokyo 106, Japan. Because of computational constraints, Protml was used only with the 4-species CS1 alignment.
 Clamped Homogeneous Electric Field (CHEF) Gel Electrophoresis: Agarose plugs were prepared by resuspending BCBL-1 cells in 1% LMP agarose (Biorad, Hercules Calif.) and 0.9% NaCl at 42° C. to a final concentration of 2.5×107 cells/ml. Solidified agarose plugs were transferred into lysis buffer (0.5M EDTA pH 8.0, 1% sarcosyl, proteinase K at 1 mg/ml final concentration) and incubated for 24 hours. Approximately 107 BCBL-1 cells were loaded in each lane. Gels were run at a gradient of 6.0 V/cm with a run time of 28 h 28 min. on a CHEF Mapper XA pulsed field gel electrophoresis apparatus (Biorad, Hercules Calif.), Southern blotted and hybridized to KS627Bam, KS330Bam and an EBV terminal repeat sequence .
 TPA Induction of Genome Replication: Late log phase BCBL-1 cells (5×105 cells per ml) were incubated with varying amounts of 12-O-tetradecanoylphorbol-13-acetate (TPA, Sigma Chemical Co., St. Louis Mo.) for 48 h, cells were then harvested and washed with phosphate-buffered saline (PBS) and DNA was isolated by chloroform-phenol extraction. DNA concentrations were determined by UV absorbance; 5 μg of whole cell DNA was quantitatively dot blot hybridized in triplicate (Manifold I, Schleicher and Schuell, Keene N.H.). KS631Bam, EBV terminal repeat and beta-actin sequences were random-primer labeled with 32P . Specific hybridization was quantitated on a Molecular Dynamics Phosphorlmager 425E.
 Cell Cultures and Transmission Studies: Cells were maintained at 5×105 cells per ml in RPMI 1640 with 20% fetal calf serum (FCS, Gibco-BRL, Gaithersburg Md.) and periodically examined for continued KSHV infection by PCR and dot hybridization. The T cell line Molt-3 (a gift from Dr. Jodi Black, Centers for Disease Control and Prevention), Raji cells (American Type Culture Collection, Rockville Md.) and RCC-1 cells were cultured in RPMI 1640 with 10% FCS. Owl monkey kidney cells (American Type Culture Collection, Rockville Md.) were cultured in MEM with 10% FCS and 1% nonessential amino acids (Gibco-BRL, Gaithersburg Md.).
 To produce the RCC-1 cell line, 2×106 Raji cells were cultivated with 1.4×106 BCBL-1 cells in the presence of 20 ng/ml TPA for 2 days in chambers separated by Falcon 0.45 μg filter tissue culture inserts to prevent contamination of Raji with BCBL-1. Demonstration that RCC-1 was not contaminated with BCBL-1 was obtained by PCR typing of HLA-DR alleles  (Raji and RCC-1: DRβ1*0310, DRβ3*02; BCBL-1: DRβ104,*07, Drβ4*01) and confirmed by flow cytometry to determine the presence (Raji, RCC1) or absence (BCBL-1) of EMA membrane antigen. Clonal sublines of RCC-1 were obtained by dilution in 96 well plates to 0.1 cells/well in RPMI 1640, 20% FCS and 30% T-STIM culture supplement (Collaborative Biomedical Products, Bedford Mass.). Subcultures were examined to ensure that each was derived from a single cluster of growing cells.
 In situ hybridization was performed with a previously described 25 bp oligomer located in ORF26 which was 5' labeled with fluorescein (Operon, Alameda Calif.) and hybridized to cytospin preparations of BCBL-1, RCC-1 and Raji cells using the methods of Lungu et al. . Slides were both directly visualized by UV microscopy and by incubating slides with anti-fluorescein-alkaline phosphatase (AP)-conjugated antibody (Boehringer-Mannheim, Indianapolis Ind.), allowing immunohistochemical detection of bound probe. Positive control hybridization was performed using a 26 bp TET-labeled EBV DNA polymerase gene oligomer (Applied Biosystems, Alameda Calif.) which was visualized by UV microscopy only and negative control hybridization was performed using a 25 bp 5' fluorescein-labeled HSV1 α47 gene oligomer (Operon, Alameda Calif.) which was visualized in a similar manner as the KSHV ORF26 probe. All nuclei of BCBL-1, RCC-1 and Raji appropriately stained with the EBV hybridization probe whereas no specific staining of the cells occurred after hybridization with the HSV1 probe.
 The remaining suspension cell lines used in transmission experiments were pelleted, and resuspended in 5 ml of 0.22 or 0.45μ filtered BCBL-1 tissue culture supernatant for 16 h. BCBL-1 supernatants were either from unstimulated cultures or from cultures stimulated with 20 ng/ml TPA. No difference in transmission to recipient cell lines was noted using various filtration or stimulation conditions. Fetal cord blood lymphocytes (FCBL) were obtained from heparinized fresh post-partum umbilical cord blood after separation on Ficoll-Paque (Pharmacia LKB, Uppsala Sweden) gradients and cultured in RPMI 1640 with 10% fetal calf serum. Adherent recipient cells were washed with sterile Hank's Buffered Salt Solution (HBSS, Gibco-BRL, Gaithersburg Md.) and overlaid with 5 ml of BCBL-1 media supernatant. After incubation with BCBL-1 media supernatant, cells were washed three times with sterile HBSS, and suspended in fresh media. Cells were subsequently rewashed three times every other day for six days and grown for at least two weeks prior to DNA extraction and testing. PCR to detect KSHV infection was performed using nested and unnested primers from ORF 26 and ORF 25 as previously described [10, 35].
 Indirect Immunofluorescence Assay: AIDS-KS sera were obtained from ongoing cohort studies (provided by Drs. Scott Holmberg, Thomas Spira and Harold Jaffe, Centers for Disease Control, and Prevention, and Isaac Weisfuse, New York City Department of Health).
 Sera from AIDS-KS patients were drawn between 1 and 31 months after initial KS diagnosis, sera from intravenous drug user and homosexual/bisexual controls were drawn after non-KS AIDS diagnosis, and sera from HIV-infected hemophiliac controls were drawn at various times after HIV infection. Immunofluorescence assays were performed using an equal volume mixture of goat anti-human IgG-FITC conjugate (Molecular Probes, Eugene Oreg.) and goat anti-human IgM-FITC conjugate (Sigma Chemical Co., St. Louis Mo.) diluted 1:100 and serial dilutions of patient sera. End-point titers were read blindly and specific immunoglobulin binding was assessed by the presence or absence of a specular fluorescence pattern in the nuclei of the plated cells. To adsorb cross-reacting antibodies, 20 μl serum diluted 1:10 in phosphate-buffer saline (PBS), pH 7.4, were adsorbed with 1-3×107 paraformaldehyde-fixed P3H3 cells for 4-10 h at 25° C. and removed by low speed centrifugation. P3H3 were induced prior to fixation with 20 ng/ml TPA for 48 h, fixed with 1% paraformaldehyde in PBS for 2 h at 4° C., and washed three times in PBS prior to adsorption.
Sequence Analysis of a 20.7 kb KSHV DNA Sequence:
 To demonstrate that KS330Bam and KS631Bam are genomic fragments from a new and previously uncharacterized herpesvirus, a lambda phage clone (KS5) derived from an AIDS-KS genomic DNA library was identified by hybridization to the KS330Bam sequence. The KS5 insert was subcloned after NotI/BamHI digestion into five subfragments and both strands of each fragment were sequenced by primer walking or nested deletion with a 7-fold average redundancy. The KS5 sequence is 20,705 bp in length and has a G+C content of 54.0%. The observed/expected CpG dinucleotide ratio is 0.92 indicating no overall CpG suppression in this region.
 Open reading frame (ORF) analysis identified 15 complete ORFs with coding regions ranging from 231 bp to 4128 bp in length, and two incomplete ORFs at the termini of the KS5 clone which were 135 and 552 by in length (FIG. 12). The coding probability of each ORF was analyzed using GRAIL 2 and CodonPreference which identified 17 regions having excellent to good protein coding probabilities. Each region is within an ORF encoding a homolog to a known herpesvirus gene with the exception of one ORF located at the genome position corresponding to ORF28 in herpesvirus saimiri (HVS). Codon preference values for all of the ORFs were higher across predicted ORFs than in non-coding regions when using a codon table composed of KS5 homologs to the conserved herpesvirus major capsid (MCP), glycoprotein H (gH), thymidine kinase (TK), and the putative DNA packaging protein (ORF29a/ORF29b) genes.
 The translated sequence of each ORF was used to search GenBank/EMBL databases with BLASTX and FastA algorithms [2, 38]. All of the putative KS5 ORFs, except one, have sequence and collinear positional homology to ORFs from gamma-2 herpesviruses, especially HVS and equine herpesvirus 2 (EHV2). Because of the high degree of collinearity and amino acid sequence similarity between KSHV and HVS, KSHV ORFs have been named according to their HVS positional homologs (i.e. KSHV ORF25 is named after HVS ORF 25).
 The KS5 sequence spans a region which includes three of the seven conserved herpesvirus gene blocks (FIG. 14) . ORFs present in these blocks include genes which encode herpesvirus virion structural proteins and enzymes involved in DNA metabolism and replication. Amino acid identities between KS5 ORFs and HVS ORFs range from 30% to 60%, with the conserved MCP ORF25 and ORF29b genes having the highest percentage amino acid identity to homologs in other gammaherpesviruses. KSHV ORF28, which has no detectable sequence homology to HVS or EBV genes, has positional homology to HVS ORF28 and EBV BDLF3. ORF28 lies at the junction of two gene blocks (FIG. 14); these junctions tend to exhibit greater sequence divergence than intrablock regions among herpesviral genomes . Two ORFs were identified with sequence homology to the putative spliced protein packaging genes of HVS (ORF29a/ORF29b) and herpes simplex virus type 1 (UL15). The KS330Bam sequence is located within KSHV ORF26, whose HSV-1 counterpart, VP23, is a minor virion structural component.
 For every KSHV homolog, the HVS amino acid similarity spans the entire gene product, with the exception of ORF21, the TK gene. The KSHV TK homolog contains a proline-rich domain at its amino terminus (nt 20343-19636; as 1-236) that is not conserved in other herpesvirus TK sequences, while the carboxyl terminus (nt 19637-18601; as 237-565) is highly similar to the corresponding regions of HVS, EHV2, and bovine herpesvirus 4 (BHV4) TK. A purine binding motif with a glycine-rich region found in herpesviral TK genes, as well as other TK genes, is present in the KSHV TK homolog (GVMGVGKS; as 260-267).
 The KS5 translated amino acid sequences were searched against the PROSITE Dictionary of Protein Sites and Patterns (Dr. Amos Bairoch, University of Geneva. Switzerland) using the computer program Motifs. Four sequence motif matches were identified among KSHV hypothetical protein sequences. These matches included: (i) a cytochrome c family heme-binding motif in ORF33 (CVHCHG; as 209-214) and ORF34 (CLLCHI; as 257-261), (ii) an immunoglobulin and major histocompatibility complex protein signature in ORF25 (FICQAKH; as 1024-1030), (iii) a mitochondrial energy transfer protein motif in ORF26 (PDDITRMRV; as 260-268), and (iv) the purine nucleotide binding site identified in ORF21. The purine binding motif is the only motif with obvious functional significance. A cytosine-specific methylase motif present in HVS ORF27 is not present in KSHV ORF27. This motif may play a role in the methylation of episomal DNA in cells persistently infected with HVS .
 Phylogenetic Analysis of KSHV: Amino acid sequences translated from the KS5 sequence were aligned with corresponding sequences from other herpesviruses. On the basis of the level of conserved aligned residues and the low incidence of introduced gaps, the amino acid alignments for ORFs 21, 22, 23, 24, 25, 26, 29a, 29b, 31 and 34 were suitable for phylogenetic analyses.
 To demonstrate the phylogenetic relationship of KSHV to other herpesviruses, a single-gene comparison was made for ORF25 (MCP) homologs from KS5 and twelve members of Herpesviridae (FIGS. 15A-15B). The thirteen available MCP amino acid sequences are large (1376 a.a. residues for the KSHV homolog) and alignment required only a low level of gapping; however, the overall similarity between viruses is relatively low . The MCP set gave stable trees with high bootstrap scores and assigned the KSHV homolog to the gamma-2 sublineage (genus Rhadinovirus), containing HVS, EHV2 and BVH4 [20, 33, 43]. KSHV was most closely associated with HVS. Similar results were obtained for single-gene alignments of TK and UL15/ORF29 sets but with lower bootstrap scores so that among gamma-2 herpesvirus members branching orders for EHV2, HVS and KSHV were not resolved.
 To determine the relative divergence between KSHV and other gammaherpesviruses, alignments for the nine genes listed above were concatenated to produce a combined gammaherpesvirus gene set (CS1) containing EBV, EHV2, HVS and KSHV amino acid sequences. The total length of CS1 was 4247 residues after removal of positions containing gaps introduced by the alignment process in one or more of the sequences. The CS1 alignment was analyzed by the ML method, giving the tree shown in FIG. 15B and by the MP and NJ methods used with the aligned herpesvirus MCP sequences. All three methods identified KSHV and HVS as sister groups, confirming that KSHV belongs in the gamma-2 sublineage with HVS as its closest known relative. It was previously estimated that divergence of the HVS and EHV2 lineages may have been contemporary with divergence of the primate and ungulate host lineages . The results for the CS1 set suggest that HVS and KSHV represent a lineage of primate herpesviruses and, based on the distance between KSHV and HVS relative to the position of EHV2, divergence between HVS and KSHV lines is ancient.
GenomiC Studies of KSHV:
 CHEF electrophoresis performed on BCBL-1 cells embedded in agarose plugs demonstrated the presence of a nonintegrated KSHV genome as well as a high molecular weight species (FIGS. 16A-16B). KS631Bam (FIG. 16A) and KS33 Bam specifically hybridized to a single CHEF gel band comigrating with 270 kilobase (kb) linear DNA standards. The majority of hybridizing DNA was present in a diffuse band at the well origin; a low intensity high molecular weight (HMW) band was also present immediately below the origin (FIG. 16A. arrow). The same filter was stripped and probed with an EBV terminal repeat sequence  yielding a 150-160 kb band (FIG. 16B) corresponding to linear EBV DNA . The HMW EBV band may correspond to either circular or concatemeric EBV DNA .
 The phorbol ester TPA induces replication-competent EBV to enter a lytic replication cycle . To determine if TPA induces replication of KSHV and EBV in BCBL-1 cells, these cells were incubated with varying concentrations of TPA for 48 h (FIG. 17). Maximum stimulation of EBV occurred at 20 ng/ml TPA which resulted in an eight-fold increase in hybridizing EBV genome. Only a 1.3-1.4 fold increase in KSHV genome abundance occurred after 20-80 ng/ml TPA incubation for 48 h.
 Prior to determining that the agent was likely to be a member of Herpesviridae by sequence analysis, BCBL-1 cells were cultured with Raji cells, a nonlytic EBV transformed B cell line, in chambers separated by a 0.45μ tissue culture filter. Recipient Raji cells generally demonstrated rapid cytolysis suggesting transmission of a cytotoxic component from the BCBL-1 cell line. One Raji line cultured in 10 ng/ml TPA for 2 days, underwent an initial period of cytolysis before recovery and resumption of logarithmic growth. This cell line (RCC-1) is a monoculture derived from Raji uncontaminated by BCBL-1 as determined by PCR amplification of HLA-DR sequences.
 RCC-1 has remained positive for the KS330233 PCR product for >6 months in continuous culture (approximately 70 passages), but KSHV was not detectable by dot or Southern hybridization at any time. In situ hybridization, however, with a 25 bp KSHV ORF26-derived oligomer was used to demonstrate persistent localization of KSHV DNA to RCC-1 nuclei. As indicated in FIGS. 18A-18C, nuclei of BCBL-1 and RCC-1 (from passage ˜65) cells had detectable hybridization with the ORF26 oligomer, whereas no specific hybridization occurred with parental Raji cells (FIG. 18B). KSHV sequences were detectable in 65% of BCBL-1 and 2.6% of RCC-1 cells under these conditions. In addition, forty-five monoclonal cultures were subcultured by serial dilution from RCC-1 at passage 50, of which eight (18%) clones were PCR positive by KS330233. While PCR detection using unnested KS330233 primer pairs was lost by passage 15 in each of the clonal cultures, persistent KSHV genome was detected in 5 clones using two more sensitive nonoverlapping nested PCR primer sets  suggesting that KSHV genome is lost over time in RCC-1 and its clones.
 Low but persistent levels of KS330233 PCR positivity were found for one of four Raji, one of four Bjab, two of three Molt-3, one of one owl monkey kidney cell lines and three of eight human fetal cord blood lymphocyte (FCBL) cultures after inoculation with 0.2-0.45μ filtered BCBL-1 supernatants. Among the PCR positive cultures, PCR detectable genome was lost after 2-6 weeks and multiple washings. Five FCBL cultures developed cell clusters characteristic of EBV immortalized lymphocytes and were positive for EBV by PCR using EBER primers [23); three of these cultures were also initially KS330233 positive. None of the recipient cell lines had detectable KSHV genome by dot blot hybridization.
 Indirect immunofluorescence antibody assays (IFA) were used to assess the presence of specific antibodies against the KSHV- and EBV-infected cell line BHL-6 in the sera from AIDS-KS patients and control patients with HIV infection or AIDS. BHL-6 was substituted for BCBL-1 for reasons of convenience; preliminary studies showed no significant differences in IFA results between BHL-6 and BCBL-1. BHL-6 have diffuse immunofluorescent cell staining with most KS patient and control unabsorbed sera suggesting nonspecific antibody binding (FIGS. 19A-19D). After adsorption with paraformaldehyde-fixed, TPA-induced P3H3 (an EBV producer sublime of P3J-HR1, a gift of Dr. George Miller) to remove cross-reacting antibodies against EBV and lymphocyte antigens, patient sera generally showed specular nuclear staining at high titers while this staining pattern was absent from control patient sera (FIGS. 19B and 19D). Staining was localized primarily to the nucleus but weak cytoplasmic staining was also present at low sera dilutions.
 With unadsorbed sera, the initial endpoint geometric mean titers (GMT) against BHL-6 cell antigens for the sera from AIDS-KS patients (GMT=1:1153, range: 1:150 to 1:12,150) were higher than for sera from control, non-KS patients (GMT=1:342; range 1:50 to 1:12,150; p=0.04) (FIG. 13). While AIDS-KS patients and HIV-infected gay/bisexual and intravenous drug user control patients had similar endpoint titers to BHL-6 antigens (GMT=1:1265 and GMT=1:1578, respectively), hemophilic AIDS patient titers were lower (GMT=1:104). Both case and control patient groups had elevated IFA titers against the EBV infected cell line P3H3.
 The difference in endpoint GMT between case and control titers against HBL-6 antigens increased after adsorption with P3H3. After adsorption, case GMT declined to 1:780 and control GMT declined to 1:81 (p=0.00009). Similar results were obtained by using BCBL-1 instead of HBL-6 cells, by pre-adsorbing with EBV-infected nonproducer Raji cells instead of P3H3 and by using sera from a homosexual male KS patient without HIV infection, in complete remission for KS for 9 months (BHL-6 titer 1:450, P3H3 titer 1:150). Paired sera taken 8-14 months prior to KS onset and after KS onset were available for three KS patients: KS patients 8 and 13 had eight-fold rises and patient 8 had a three-fold fall in P3H3-adsorbed BCBL-1 titers from pre-onset sera to post-KS sera.
 These studies demonstrate that specific DNA sequences found in KS lesions by representational difference analysis belong to a newly identified human herpesvirus. The current studies define this agent as a human gamma-2 herpesvirus that can be continuously cultured in naturally-transformed, EBV-coinfected lymphocytes from AIDS-related body-cavity based lymphomas.
 Sequence analysis of the KS5 lambda phage insert provides clear evidence that the KS330Bam sequence is part of a larger herpesvirus genome. KS5 has a 54.0% G+C content which is considerably higher than the corresponding HVS region (34.3% G+C). While there is no CpG dinucleotide suppression in the KS5 sequence, the corresponding HVS region has a 0.33 expected:observed CpG dinucleotide ratio . The CpG dinucleotide frequency in herpesviruses varies from global CpG suppression among gammaherpesviruses to local CpG suppression in the betaherpesviruses, which may result from deamination of 5'-methylcytosine residues at CpG sites resulting in TpG substitutions . CpG suppression among herpesviruses [21, 30, 44] has been hypothesized to reflect co-replication of latent genome in actively dividing host cells, but it is unknown whether or not KSHV is primarily maintained by a lytic replication cycle in vivo.
 The 20,705 bp KS5 fragment has 17 protein-coding regions, 15 of which are complete ORFs with appropriately located TATA and polyadenylation signals, and two incomplete ORFs located at the phage insert termini. Sixteen of these ORFs correspond by sequence and collinear positional homology to 15 previously identified herpesviral genes including the highly conserved spliced gene. The conserved positional and sequence homology for KSHV genes in this region are consistent with the possibility that the biological behavior of the virus is similar to that of other gammaherpesviruses. For example, identification of a thymidine kinase-like gene on KS5 implies that the agent is potentially susceptible to TK-activated DNA polymerase inhibitors and like other herpesviruses possesses viral genes involved in nucleotide metabolism and DNA replication . The presence of major capsid protein and glycoprotein H gene homologs suggest that replication competent virus would produce a capsid structure similar to other herpesviruses.
 Phylogenetic analyses of molecular sequences show that KSHV belongs to the gamma-2 sublineage of the Gammaherpesvirinae subfamily, and is thus the first human gamma-2 herpesvirus identified. Its closest known relative based on available sequence comparisons is HVS, a squirrel monkey gamma-2 herpesvirus that causes fulminant polyclonal T cell lymphoproliferative disorders in some New World monkey species. Data for the gamma-2 sublineage are sparse: only three viruses (KSHV, HVS and EHV2) can at present be placed on the phylogenetic tree with precision (the sublineage also contains murine herpesvirus 68 and BHV4 ). Given the limitation in resolution imposed by this thin background, KSHV and HVS appear to represent a lineage of primate gamma-2 viruses. Previously, McGeoch et al.  proposed that lines of gamma-2 herpesviruses may have originated by cospeciation with the ancestors of their host species. Extrapolation of this view to KSHV and HVS suggests that these viruses diverged at an ancient time, possibly contemporaneously with the divergence of the Old World and New World primate host lineages. Gammaherpesviruses are distinguished as a subfamily by their lymphotrophism  and this grouping is supported by phylogenetic analysis based on sequence data . The biologic behavior of KSHV is consistent with its phylogenetic designation in that KSHV can be found in in vitro lymphocyte cultures and in in vivo samples of lymphocytes .
 This band appears to be a linear form of the genome because other "high molecular weight" bands are present for both EBV and KSHV in BCBL-1 which may represent circular forms of their genomes. The linear form of the EBV genome, associated with replicating and packaged DNA  migrates substantially faster than the closed circular form associated with latent viral replication . While the 270 kb band appears to be a linear form, it is also consistent with a replicating dimer plasmid since the genome size of HVS is approximately 135 kb. The true size of the genome may only be resolved by ongoing mapping and sequencing studies.
 Replication deficient EBV mutants are common among EBV strains passaged through prolonged tissue culture . The EBV strain infecting Raji, for example, is an BALF-2 deficient mutant ; virus replication is not inducibile with TPA and its genome is maintained only as a latent circular form [23, 33]. The EBV strain coinfecting BCBL-1 does not appear to be replication deficient because TPA induces eight-fold increases in DNA content and has an apparent linear form on CHEF electrophoresis. KSHV replication, however, is only marginally induced by comparable TPA treatment indicating either insensitivity to TPA induction or that the genome has undergone loss of genetic elements required for TPA induction. Additional experiments, however, indicate that KSHV DNA can be pelleted by high speed centrifugation of filtered organelle-free, DNase I-protected BCBL-1 cell extracts, which is consistent with KSHV encapsidation.
 Transmission of KSHV DNA from BCBL-1 to a variety of recipient cell lines is possible and KSHV DNA can be maintained at low levels in recipient cells for up to 70 passages. However, detection of virus genome in recipient cell lines by PCR may be due to physical association of KSHV DNA fragments rather than true infection. This appears to be unlikely given evidence for specific nuclear localization of the ORF26 sequence in RCC-1. If transmission of infectious virus from BCBL-1 occurs, it is apparent that the viral genome declines in abundance with subsequent passages of recipient cells. This is consistent with studies of spindle cell lines derived from KS lesions. Spindle cell cultures generally have PCR detectable KSHV genome when first explanted, but rapidly lose viral genome after initial passages and established spindle cell cultures generally do not have detectable KSHV sequences .
 Infections with the human herpesviruses are generally ubiquitous in that nearly all humans are infected by early adulthood with six of the seven previously identified human herpesviruses . Universal infection with EBV, for example, is the primary reason for the difficulty in clearly establishing a causal role for this virus in EBV-associated human tumors. The serologic studies identified nuclear antigen in BCBL-1 and HBL-6 which is recognized by sera from AIDS-KS patients but generally not by sera from control AIDS patients without KS after removal of EBV-reactive antibodies. These data are consistent with PCR studies of KS and control patient lymphocytes suggesting that KSHV is not ubiquitous among adult humans, but is specifically associated with persons who develop Kaposi's sarcoma. In this respect, it appears to be epidemiologically similar to HSV2 rather than the other known human herpesviruses. An alternative possibility is that elevated IFA titers against BCBL-1 reflect disease status rather than infection with the virus.
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EXPERIMENTAL DETAILS SECTION III
 KS Patient Enrollment: Cases and controls were selected from ongoing cohort studies based on the availability of clinical information and appropriate PBMC samples. 21 homosexual or bisexual men with AIDS who developed KS during their participation in prospective cohort studies were identified [14-16]. Fourteen of these patients had paired PBMC samples collected after KS diagnosis (median +4 months) and at least four months prior to KS diagnosis (median -13 months), while the remaining 7 had paired PBMC taken at the study visit immediately prior to KS diagnosis (median -3 months) and at entry into their cohort study (median -51 months prior to KS diagnosis).
 Hemophilic and Homosexual/Bisexual Male AIDS Patient Control Enrollment: Two control groups of AIDS patients were examined: 23 homosexual/bisexual men with AIDS followed until death who did not develop KS ("high risk" control group) from the Multicenter AIDS Cohort Study ), and 19 hemophilic men ("low risk" control group) enrolled from joint projects of the National Hemophilia Foundation and the Centers for Disease Control and Prevention. Of the 16 hemophilic controls with available follow-up information, none are known to have developed KS and <2% of hemophilic AIDS patients historically develop KS . For homosexual/bisexual AIDS control patients who did not develop KS, paired PBMC specimens were available at entry into their cohort study (median -35 months prior to AIDS onset) and at the study visit immediately prior to nonKS AIDS diagnosis (median HBL-6 months prior to AIDS onset).
 DNA Extraction and Analyses: DNA from 106-107 PBMC in each specimen was extracted and quantitated by spectrophotometry. Samples were prepared in physically isolated laboratories from the laboratory where polymerase chain reaction (PCR) analyses were performed.
 All samples were tested for amplifiability using primers specific for either the HLA-DQ locus (GH26/GH27) or b-globin . PCR detection of KSHV DNA was performed as previously described  with the following nested primer sets: No. 1 outer 5'-AGCACTCGCAGGGCAGTACG-3' (SEQ ID NO:51), 5'-GACTCTTCGCTGATGAACTGG-3' (SEQ ID NO:52); No. 1 inner 5'-TCCGTGTTGTCTACGTCCAG-3' (SEQ ID NO:53), 5'-AGCCGAAAGGATTCCACCAT-3' (SEQ ID NO:41); No. 2 outer 5'-AGGCAACGTCAGATGTGAC-3' (SEQ ID NO:54), 5'-GAAATTACCCACGAGATCGC-3' (SEQ ID NO:42); No. 2 inner 5'-CATGGGAGTACATTGTCAGGACCTC-3' NO:55), (SEQ ID 5'-GGAATTATCTCGCAGGTTGCC-3' (SEQ ID NO:56); No. 3 outer 5'-GGCGACATTCATCAACCTCAGGG-3' (SEQ ID NO:57), 5'-ATATCATCCTGTGCGTTCACGAC-3' (SEQ ID NO:58); No. 3 inner 5'-CATGGGAGTACATTGTCAGGACCTC-3' (SEQ ID NO: 55), 5'-GGAATTATCTCGCAGGTTGCC-3' (SEQ ID NO:56). The outer primer set was amplified for 35 cycles at 94° C. for 30 seconds, 60° C. for 1 minute and 72° C. for 1 minute with a 5 minute final extension cycle at 72° C. One to three ml of the PCR product was added to the inner PCR reaction mixture and amplified for 25 additional cycles with a 5 minute final extension cycle. Primary determination of sample positivity was made with primer set No. 1 and confirmed with either primer sets 2 or 3 which amplify nonoverlapping regions of the KSHV hypothetical major capsid gene. Sampling two portions of the KSHV genome decreased the likelihood of intraexperimental PCR contamination. These nested primer sets are 2-3 logs more sensitive for detecting KSHV sequences than the previously published KS330233 primers  and are estimated to be able to detect <10 copies of KSHV genome under optimal conditions. Sample preparations were prealiquoted and amplified with alternating negative control samples without DNA to monitor and control possible contamination. All samples were tested in a blinded fashion and a determination of the positivity/negativity made before code breaking. 35 Significance testing was performed with Mantel-Haenszel chi-squared estimates and exact confidence intervals using Epi-Info ver. 6 (USD Inc., Stone Mt. GA).
KSHV Positivity of Case and Control PBMC Samples:
 Paired PBMC samples were available from each KS patient and homosexual/bisexual control patient; a single sample was available from each hemophilic control patient.
 To determine the KSHV positivity rate for each group of AIDS patients, a single specimen from each participant taken closest to KS or other AIDS-defining illness ("second sample") was analyzed. Overall, 12 of 21 (57%) of PBMC specimens from KS patients taken from 6 months prior to KS diagnosis to 20 months after KS diagnosis were KSHV positive. There was no apparent difference in positivity rate between immediate pre-diagnosis and post-diagnosis visit specimens (4 of 7 (57%) vs. 8 of 14 (57%) respectively).
 The number of KSHV positive control PBMC specimens from both homosexual/bisexual (second visit) and hemophilic patient controls was significantly lower. Only 2 of 19 (11%) hemophilic PBMC samples were positive (odds ratio 11.3, 95% confidence interval 1.8 to 118) and only 2 of 23 (9%) PBMC samples from homosexual/bisexual men who did not develop KS were positive (odds ratio 14.0, 95% confidence interval 2.3 to 144). If all KS patient PBMC samples taken immediately prior to or after diagnosis were truly infected, the PCR assay was at least 57% sensitive in detecting KSHV infection among PBMC samples. No significant differences in CD4+ counts were found for KS patients and homosexual/bisexual patients without KS at the second sample evaluation (Kruskall-Wallis p=0.15) (FIG. 21). CD4+ counts from the single sample from hemophilic AIDS patients were higher than CD4+ counts from KS patients (Kruskall-Wallis p=0.004), although both groups showed evidence of HIV-related immunosuppression.
 Paired specimens were available from all 21 KS patients and 23 homosexual/bisexual male AIDS control patients who did not develop KS. For the KS group, initial PBMC samples were taken four to 87 months (median 13 months) prior to the onset of KS. Initial PBMC samples from the control group were drawn 13 to 106 months (median 55 months) prior to onset of first nonKS AIDS-defining illness (1987 CDC surveillance definition). 11 of 21 (52%) of KS patients had detectable KSHV DNA in PBMC samples taken prior to KS onset compared to 2 of 19 (11%, p=0.005) hemophilic control samples, and 1 (4%, p=0.0004) and 2 (9%, p=0.002) of 23 homosexual/bisexual control samples taken at the first and second visits respectively (FIGS. 20A-20B). The figure shows that 7 of the paired KS patient samples were positive at both visits, 5 KS patients and 2 control patients converted from negative to positive and two KS patients and one control patient reverted from positive to negative between visits. The remaining 7 KS patients and 20 control patients were negative at both visits.
 For the 5 KS patients that converted from an initial negative PBMC result to a positive result at or near to KS diagnosis, the median length of time between the first sample and the KS diagnosis was 19 months. Three of the 6 KS patients that were negative at both visits had their last PBMC sample drawn 2-3 months prior to onset of illness. It is unknown whether these patients became infected between their last study visit and the KS diagnosis date.
 Ambroziak and coworkers have found evidence that KSHV preferentially infects CD19+ B cells by PBMC subset examination of three patients . Other gammaherpesviruses, such as Epstein-Barr virus (EBV) and herpesvirus saimiri are also lymphotrophic herpesviruses and can cause lymphoproliferative disorders in primates [11, 20].
 It is possible that KSHV, like most human herpesviruses, is a ubiquitous infection of adults . EBV, for example, is detectable by PCR in CD19+ B lymphocytes from virtually all seropositive persons  and approximately 98% MACS study participants had EBV VCA antibodies at entry into the cohort study . The findings, however, are most consistent with control patients having lower KSHV infection rates than cases and that KSHV is specifically associated with the subsequent development of KS. While it is possible that control patients are infected but have an undetectably low KSHV viral PBMC load, the inability to find evidence of infection in control patients under a variety of PCR conditions suggests that the majority of control patients are not infected. Nonetheless, approximately 10% of these patients were KSHV infected and did not develop KS. It is unknown whether or not this is similar to the KSHV infection rate for the general human population.
 This study demonstrates that KSHV infection is both strongly associated with KS and precedes onset of disease in the majority of patients. 57% of KS patients had detectable KSHV infection at their second follow-up visit (52% prior to the onset of KS] compared to only 9% of homosexual/bisexual (p=0.002) and 11% of hemophilic control patients (p=0.005). Despite similar CD4+ levels between homosexual/bisexual KS cases and controls, KSHV DNA positivity rates were significantly higher for cases at both the first (p=0.005) and second sample visits indicating that immunosuppression alone was not responsible for these elevated detection rates. It is also unlikely that KSHV simply colonizes existing KS lesions in AIDS patients since neither patient group had KS at the time the initial sample was obtained. Five KS patients and two homosexual/bisexual control patients converted from a negative to a positive, possibly due to new infection acquired during the study period.
 The findings are in contrast to PCR detection of KSHV DNA in all 10 PBMC samples from KS patients by Ambroziak at al. . It is possible that the assay was not sensitive enough to detect virus in all samples since it was required that each positive sample to be repeatedly positive by two independent primers in blinded PCR assays. This appears unlikely, however, given the sensitivity of the PCR nested primer sets. The 7 KS patients who were persistently negative on both paired samples may represent an aviremic or low viral load subpopulation of KS patients. The PCR conditions test a DNA amount equivalent to approximately 2×103 lymphocytes; an average viral load less than 1 copy per 2×103 cells may be negative in the assay. Two KS patients and a homosexual/bisexual control patient initially positive for KSHV PCR amplification reverted to negative in samples drawn after diagnosis. These results probably reflect inability to detect KSHV DNA in peripheral blood rather than true loss of infection although more detailed studies of the natural history of infection are needed.
 The study was designed to answer the fundamental question of whether or not infection with KSHV precedes development of the KS phenotype. The findings indicate that there is a strong antecedent association between KSHV infection and KS. This temporal relationship is an absolute requirement for establishing that KSHV is central to the causal pathway for developing KS. This study contributes additional evidence for a possible causal role for this virus in the development of KS.
 1. Katz M H, Hessol N A, Buchbinder S P, Hirozawa A, O'Malley P, Holmberg S D. Temporal trends of opportunistic infections and malignancies in homosexual men with AIDS. J Infect Dis. 1994; 170:198-202.  2. Beral V, Peterman T A, Berkelman R L, Jaffe H W. Kaposi's sarcoma among persons with AIDS: a sexually transmitted infection?Lancet. 1990; 335:123-128.  3. Archibald C P, Schechter M T, Le T N, Craib K J P, Montaner J S G, O'Shaughnessy M V. Evidence for a sexually transmitted cofactor for AIDS-related Kaposi's sarcoma in a cohort of homosexual men. Epidemiol. 1992; 3:203-209.  4. Beral V, Bull D, Jaffe H, Evans B, Gill N, Tillett H et al. Is risk of Kaposi's sarcoma in AIDS patients in Britain increased if sexual partners came from United States or Africa?BMJ. 1991; 302:624-5.  5. Beral V. Epidemiology of Kaposi's sarcoma. Cancer, HIV and AIDS. London: Imperial Cancer Research Fund; 1991:5-22.  6. Chang Y, Cesarman E, Pessin M S, Lee F, Culpepper J, Knowles D M, et al. Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma. Science. 1994; 265:1865-69.  7. Moore P S, Chang Y. Detection of herpesvirus-like DNA sequences in Kaposi's sarcoma lesions from persons with and without HIV infection. New England J Med. 1995; 332:1181-1185.  8. Boshoff C, Whitby D, Hatziionnou T, Fisher C, van der Walt J, Hatzakis A et al. Kaposi's sarcoma-associated herpesvirus in HIV-negative Kaposi's sarcoma. Lancet. 1995; 345:1043-44.  9. Su I-J, Hsu Y-S, Chang Y-C, Wang I-W. Herpesvirus-like DNA sequence in Kaposi's sarcoma from AIDS and non-AIDS patients in Taiwan. Lancet. 1995; 345:722-23.  10. Dupin N, Grandadam M, Calvez V, Gorin I, Aubin J T, Harvard S, et al. Herpesvirus-like DNA in patients with Mediterranean Kaposi's sarcoma. Lancet. 1995; 345:761-2.  11. Miller G. Oncogenicity of Epstein-Barr virus. J Infect Dis. 1974; 130:187-205.  12. Hill A B. Environment and disease: association or causation? Proc Roy Soc Med. 1965; 58:295-300.  13. Susser M. Judgment and causal inference: criteria in epidemiologic studies. An J. Epid. 1977; 105:1-15.  14. Fishbein D B, Kaplan J E, Spira T J, Miller B, Schonberger L B, Pinsky P F, et al. Unexplained lymphadenopathy in homosexual men: a longitudinal study. JAMA. 1985; 254:930-5.  15. Holmberg S D. Possible cofactors for the development of AIDS-related neoplasms. Cancer Detection and Prevention. 1990; 14:331-336.  16. Kaslow R A, Ostrow D G, Detels R, Phair J P, Polk B F, Rinaldo C R. The Multicenter AIDS Cohort Study: rationale, organization and selected characteristics of the participants. Am J Epidemiol. 1987; 126:310-318.  17. Wolinsky S. Rinaldo C, Kwok S, Sinsky J, Gupta P. Imagawa D, et al. Human immunodeficiency virus type 1 (HIV-1) infection a median of 18 months before a diagnostic Western blot. Ann Internal Med. 1989; 111:961.  18. Bauer H M, Ting Y, Greer C E, Chambers J C, Tashiro C J, Chimera J, et al. Genital papillomavirus infection in female university students as determined by a PCR-based method. JAMA. 1991; 265:2809-10.  19. Ambroziak J A, Blackbourn D J, Herndier B G, Glogau R G, Gullett J H, McDonald A R, at al. Herpes-like sequences in HIV-infected and uninfected Kaposi's sarcoma patients. Science. 1995; 268:582-583.  20. Roizman B. The family Herpesviridae. In: Roizman B, Whitley R J, Lopez C, eds. The Human Herpeviruses. New York: Raven Press, Ltd.; 1993:1-9.  21, Roizman B. New viral footprints in Kaposi's sarcoma. N Engl J Med. 1995; 332:1227-1228.  22. Miyashita E M, Yang B, Lam K M C, Crawford D H, Thorley-Lawson D A. A novel form of Epstein-Barr virus latency in normal B cells in vivo. Cell. 1995; 80:593-601.  23. Rinaldo C R, Kingsley L A, Lyter D W, Rabin B S, Atchison R W, Bodner A J, at al. Association of HTLV-III with Epstein-Barr virus infection and abnormalities of T lymphocytes in homosexual men. J Infect Dis. 1986; 154:556-61.
EXPERIMENTAL DETAILS SECTION IV
 To determine if the KHV-KS virus is also present in both endemic and HIV-associated KS lesions from African patients, formalin-fixed, paraffin-embedded tissues from both HIV seropositive and HIV seropositive Ugandan KS patients were compared to cancer tissues from patients without KS in a blinded case-control study.
 Patient Enrollment: Archival KS biopsy specimens were selected from approximately equal numbers of HIV-associated and endemic HIV-negative KS patients enrolled in an ongoing case-control study of cancer and HIV infection at Makerere University, Kampala Uganda. Control tissues were consecutive archival biopsies from patients with various malignancies enrolled in the same study, chosen without prior knowledge of HIV serostatus. All patients were tested for HIV antibody (measured by Cambridge Bioscience Recombigen Elisa assay).
 Tissue preparation: Each sample examined was from an individual patient. Approximately ten tissue sections were cut (10 micron) from each paraffin block using a cleaned knife blade for each specimen. Tissue sections were deparaffinized by extracting the sections twice with 1 ml xylene for 15 min. followed by two extractions with 100% ethanol for 15 min. The remaining pellet was then resuspended and incubated overnight at 50° C. in 0.5 ml of lysis buffer (25 mM KCl, 10 mM Tris-HCl, pH 8.3, 1.4 mM MgCl2, 0.01% gelatin, 1 mg/ml proteinase K). DNA was extracted with phenol/chloroform, ethanol precipitated and resuspended in 10 mM Tris-HCl, 0.1 mM EDTA, pH 8.3.
 PCR Amplification: 0.2-0.4 ug of DNA was used in PCR reactions with KS330233 primers as previously described . The samples which were negative were retested by nested PCR amplification, which is approximately 102-103 fold more sensitive in detecting KS330233 sequence than the previously published KS330233 primer set . These samples were tested twice and samples showing discordant results were retested a third time. 51 of 74 samples initially examined were available for independent extraction and testing at Chester Beatty Laboratories, London using identical nested PCR primers and conditions to ensure fidelity of the PCR results. Results from eight samples were discordant between laboratories and were removed from the analysis as uninterpretable (four positive samples from each laboratory). Statistical comparisons were made using EPI-INFO ver. 5 (USD, Stone Mt. GA, USA) with exact confidence intervals.
 Of 66 tissues examined, 24 were from AIDS-KS cases, 20 were from endemic HIV seronegative KS cases, and 22 were from cancer control patients without KS. Seven of the cancer control patients were HIV seropositive and 15 were HIV seronegative (FIG. 22). Tumors examined in the control group included carcinomas of the breast, ovaries, rectum, stomach, and colon, fibrosarcoma, lymphocytic lymphomas, Hodgkin's lymphomas, choriocarcinoma and anaplastic carcinoma of unknown primary site. The median age of AIDS-KS patients was 29 years (range 3-50) compared to 36 years (range 3-79) for endemic KS patients and 38 years (range 21-73) for cancer controls.
 Among KS lesions, 39 of 44 (89%) were positive for KS330233 PCR product, including KS tissues from 22 of 24 (92%) HIV seropositive and 17 of 20 (85%) HIV seronegative patients. In comparison, 3 of 22 (14%) nonKS cancer control tissues were positive, including 1 of 7 (14%) HIV seropositive and 2 of 15 (13%) HIV seronegative control patients (FIG. 19). These control patients included a 73 year old HIV seronegative male and a 29 year old HIV seronegative female with breast carcinomas, and a 36 year old HIV seropositive female with ovarian carcinoma. The odds ratios for detecting the sequences in tissues from HIV seropositive and HIV seronegative cases and controls was 66 (95% confidence interval (95% C.I.) 3.8-3161) and 36.8 (95% C.I. 4.3-428) respectively. The overall weighted Mantel-Haenzel odds ratio stratified by HIV serostatus was 49.2 (95% C.I. 9.1-335). KS tissues from four HIV seropositive children (ages 3, 5, 6, and 7 years) and four HIV seronegative children (ages 3, 4, 4, and 12 years) were all positive for KS330233.
 All discordant results (i.e. KSHV negative KS or KSHV positive nonKS cancers) were reviewed microscopically. All KS330233 PCR negative KS samples were confirmed to be KS. Likewise, all KS330233 PCR positive nonKS cancers were found not to have occult KS histopathologically.
 These results indicate that KSHV DNA sequences are found not only in AIDS-KS , classical KS  and transplant KS  but also in African KS from both HIV seropositive and seronegative patients. Despite differences in clinical and epidemiological features, KSHV DNA sequences are present in all major clinical subtypes of KS from widely dispersed geographic settings.
 This study was performed on banked, formalin-fixed tissues which prevented the use of specific detection assays such as Southern hybridization. DNA extracted after such treatment is often fragmented which reduces the detection sensitivity of PCR and may account for the 5 PCR negative KS samples found in the study. The results, however, are unlikely to be due to PCR contamination or nonspecific amplification. Specimens were tested blindly and a subset of samples were independently extracted and tested at a physically separate laboratory. Specimen blinding is essential to ensure the integrity of results based solely on PCR analyses. A subset of amplicons was sequenced and found to be more than 98% identical to the published KS330233 sequence confirming their specific nature and, because of minor sequence variation, making the possibility of contamination unlikely.
 In contrast to previous studies in North American and European populations, it was found 3 of 22 control tissues to have evidence of KSHV infection. Since these cancers represent a variety of tissue types, it is unlikely that KSHV has an etiologic role in these tumors. One possible explanation for the findings is that these results reflect the rate of KSHV infection in the nonKS population in Uganda. Four independent controlled studies from North America [5 and 9] Europe  and Asia  have failed to detect evidence of KSHV infection in over 200 cancer control tissues, with the exception of an unusual AIDS-associated, body-cavity-based lymphoma . Taken together, these studies indicate that DNA-based detection of KSHV infection is rare in most nonKS cancer tissues from developed countries. KSHV infection has been reported in post-transplant skin tumors, although well-controlled studies are needed to confirm that these findings are not due to PCR contamination . Since the rate of HIV-negative KS is much more frequent in Uganda than the United States, detection of KSHV in control tissues from cancer patients in the study may reflect a relatively high prevalence infection in the general Ugandan population.
 While KS is extremely rare among children in developed countries , the rate of KS in Ugandan children has risen dramatically over the past 3 decades: age-standardized rates (per 100,000) for boys age 0-14 years were 0.25 in 1964-68 and 10.1 in 1992-93. Detection of KSHV genome in KS lesions from prepubertal children suggests that the virus has a nonsexual mode of transmission among Ugandan children. That five of these children were 5 years old or less raises the possibility that the agent can be transmitted perinatally. Whether or not immune tolerance due to perinatal transmission accounts for the more fulminant form of KS occurring in African children remains to be investigated.
 1. Oettle A. G. Geographic and racial differences in the frequency of Kaposi's sarcoma as evidence of environmental or genetic causes. Acta Un Int Cancer 1962; 18:330-363.  2. Beral V. Epidemiology of Kaposi's sarcoma. In: Cancer, HIV and AIDS. London: Imperial Cancer Research Fund, 1991: 5-22.  3. Wabinga H. R., Parkin D. M., Wabwire-Mangen F., Mugerwa J. Cancer in Kampala, Uganda, in 1989-91: changes in incidence in the era of AIDS. Int J Cancer 1993; 54:26-36.  4. Kestens L. et al. Endemic Kaposi's sarcoma is not associated with immunodeficiency. Int. J. Cancer 1985; 36:49-54.  5. Chang Y. et al. Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi's sarcoma. Science 1994; 266:1865-9.  6. Moore P. S. and Chang Y. Detection of herpesvirus-like DNA sequences in Kaposi's sarcoma lesions from persons with and without HIV infection. New England J Med 1995; 332:1181-85.  7. Boshoff C. et al. Kaposi's sarcoma-associated herpesvirus in HIV negative Kaposi's sarcoma (letter). Lancet 1995; 345:1043-44.  8. Su, I.-J., Hsu, Y.-S., Chang, Y.-C., Wang, I.-W. Herpesvirus-like DNA sequence in Kaposi's sarcoma from AIDS and non-AIDS patients in Taiwan. Lancet 1995; 345: 722-3.  9. Cesarman E., Chang Y. Moore P. S. Said J. W., Knowles D. M. Kaposi's sarcoma-associated herpesvirus-like DNA sequences are present in AIDS-related body cavity based lymphomas. New England J Med 1995; 332:1186-1191.  10. Rady P. L., et al. Herpesvirus-like DNA sequences in nonKaposi's sarcoma skin lesions of transplant patients. Lancet 1995; 345:1339-40.
EXPERIMENTAL DETAILS SECTION V
Serologic Marker for KSHV Infection
 Patients Serum was collected from a convenience sample of 89 HIV-infected patients seen at several clinical sites in Connecticut, New York, and California. Demographic and clinical information was recorded on standardized forms which were linked to samples by a numerical code. Patients were classified as having KS if the diagnosis was histologically confirmed or, in the opinion of the primary clinician, the diagnosis of KS was unequivocal on clinical grounds. Eighty six (97%) were male; 90 of the 86 men (93%) were homosexual or bisexual. Forty seven patients, all male, had KS. The characteristics of the study population are found in FIG. 23].
 Cell lines The BCBL-1 line was established from an AIDS-associated body cavity B cell non-Hodgkin's lymphoma . Neither BCBL-1 cells, nor the tumor from which they were derived, express surface immunoglobulin or B cell specific surface markers; however BCBL-1 cells contain immunoglobulin gene rearrangements that are characteristic of B cells . KSHV DNA sequences can be detected in BCBL-1 cells by DNA representational difference analysis [23,32]. BCBL-1 cells also contain an EBV genome detectable with several different EBV DNA probes. B95-8 is an EBV producer marmoset cell line that can be efficiently induced into EBV lytic cycle gene expression by phorbol esters (TPA) [33,34], HH 514-16 is an EBV containing cell line, originally from a Burkitt lymphoma, that is optimally inducible into EBV lytic cycle gene expression by n-butyrate [35,36]. B141 is an EBV-negative Burkitt lymphoma cell line . B95-8, HH514-16 and BL41 do not hybridize with the KSHV probes. All cell lines were cultured in RPMI 1640 medium containing 8% fetal calf serum.
 Immunoblotting Assays Extracts of uninduced BCBL-1 cells or BCBL-1 cells that had been treated with 20 ng/ml TPA and 3 mM n-butyrate for 48 hrs were prepared by sonication. HH514-16 cells, treated similarly, served to control for antibody reactivity to EBV polypeptides. Each lane of a 10% or 12% polyacrylamide gel was loaded with extract of 5×105 cells in SDS sample buffer; electrophoresis, transfer to nitrocellulose and blocking with skim milk followed standard protocols . Sera were screened at 1:100 dilution. The reaction was developed by 1.0 μCi of 125I Staphylococcal protein A. Radioautographs were exposed to film for 24-48 hrs. Immunoblotting assays were performed and interpreted on coded sera.
 Immunofluorescent assay The antigens were BCBL-1 cells that were untreated or treated with 3 mM n-butyrate for 48 hrs. Cells were dropped onto slides that were fixed in acetone and methanol. Sera were tested at 1:10 dilution, followed by 1:30 dilution of fluoresceinated goat anti-human Ig. The reactivity of a serum was compared on untreated and n-butyrate treated BCBL-1 cells. Reactivity with 30-50% of the chemically treated BCBL-1 cells was considered a positive reaction. All immunofluorescence tests were performed on coded sera. The two readers were blinded to disease status or results of immunoblotting assays.
 Chemical Induction of lytic cycle KSHV proteins in BCBL cells: Initial experiments using the immunoblotting technique were designed to determine whether BCBL-1 cells expressed unique antigenic polypeptides that might be specific for KSHV infection. Since sera from HIV-1 infected patients with or without KS would be expected to contain antibodies to EBV polypeptides and since BCBL-1 cells are dually infected with KSHV and EBV it was essential to distinguish EBV polypeptides from those encoded or induced by KSHV. FIGS. 27A-27B, an immunoblot prepared from BCBL-1 cells reacted with a reference EBV antiserum, shows that BCBL-1 cells expressed two polypeptides, representing the latent nuclear antigen EBNA1 and p21, a late antigen complex , that were present in other EBV producer cell lines, such as B95-8 (FIG. 27A) and HH514-16 (FIG. 278 and FIGS. 28A-28D). When sera from patients with KS were used as a source of antibody they failed to identify in extracts from untreated BCBL-1 cells additional antigenic polypeptides that were not also seen in the EBV producer cell lines. However, if extracts were prepared from BCBL-1 cells that had first been treated with a combination of phorbol ester, TPA, and n-butyrate, KS patient sera now recognized a number of novel polypeptides that were present in the BCBL-1 cell line but not in standard EBV producer cell lines (FIG. 27B). The molecular weights of the most prominent of these many polypeptides were estimated at about 27 KDa, 40 KDa and 60 KDa on 10% polyacrylamide gels. These polypeptides were detected within 24 hrs after addition of the chemical inducing agents, but were not evident in BCBL-1 held in culture for as long as 5 days without chemical treatment. Further experiments showed that n-butyrate was the chemical agent primarily responsible for induction of p40, whereas p60 could be induced by TPA or n-butyrate (FIGS. 28A-28D). Since p27, p40 and p60 were not detected in untreated cells and appeared after treatment with chemicals they likely represented lytic cycle rather than latent cycle polypeptides of KSHV.
 p40 and p60 are KSHV specific: FIGS. 27A-27B shows that antigenic polypeptides corresponding in molecular weight to p40 were not observed in two EBV producer lines, B95-8 and HH514-16, that were induced into the EBV lytic cycle by the same chemicals or in comparably treated EBV-negative BL41 cells. Furthermore n-butyrate strongly induced expression of p40 in BCBL-1 cells but had little or no effect on the level of expression of the EBV p21 complex in the same cells. In related experiments it was found that n-butyrate also induced an increase in the abundance of KSHV DNA and KSHV lytic cycle mRNA. TPA, by contrast, induced the EBV lytic cycle efficiently` treatment with TPA caused an increase in the abundance of the EBV p21 protein and minimal induction of KSHV p40. These findings suggested that latency to lytic cycle switch of the two gamma herpes viruses carried by BCBL-1 cells was under separate control and that the p40 complex was specific to the KSHV genome.
 p40 as a serologic marker for KSHV: While a few highly reactive sera, such as KS 01-03, (FIG. 27B) recognized multiple antigenic proteins unique to the chemically induced BCBL-1 cells, including p27, p60 and p40, sera from other patients with KS did not react with p27 or p60 but still recognized p40 (FIGS. 28A and 28B). Therefore recognition of p40 was investigated as a serologic marker for infection with KSHV. Sera from 89 HIV-1 infected patients from Connecticut, New York and California were examined for presence of antibodies to p40; only 3 of 42 patients (7%) without KS had antibodies to p40 (p<0.0001 by Chi square). These three patients were homosexual or bisexual men from New York city. The positive and negative predictive values of the serologic marker for the presence of KS were 84% and 78% respectively. Three HIV-1 infected men from New York with non-Hodgkin's lymphoma but without KS were non-reactive to the KSHV p40 antigen. FIG. 25 compares the patients with KS whose serum did or did not contain antibodies to KSHV p40. Neither CD4 cell number nor the extent of KS disease predicted the presence or absence of a serologic response to p40.
 Immunofluorescence assays: Immunoblots showed that n-butyrate induced expression of KSHV lytic cycle polypeptides in BCBL-1 cells without significantly affecting expression of EBV polypeptides (FIG. 28A). Therefore it was reasoned that n-butyrate might also induce many more BCBL-1 cells into the KSHV lytic cycle than into the EBV lytic cycle. Using indirect immunofluorescence with a reference human antiserum, R Ml in FIG. 27B, that contains antibodies to EBV but not KSHV there were about 2% antigen positive untreated BCBL-1 cells and a similar number of antigen positive BCBL cell that had been treated with n-butyrate. Serum 01-03 that is EBV-positive and KS-positive (FIG. 27B) detected 2% antigen positive cells in the untreated BCBL population, presumably the EBV expressing cells, while it detected 50% antigen positive BCBL-1 cells that had been treated with n-butyrate. This increase in the number of antigen positive BCBL-1 cells among the n-butyrate treated population served as the basis of an immunofluorescence screening assay for antibodies to KSHV lytic cycle antigens (FIGS. 29A-29F). The results of the immunofluorescence assay were nearly identical to the immunoblotting assay (FIG. 26). Among 89 sera there were only 4 (3%) that were discordant in the two assays. Three sera scored positive by IFA and negative by immunoblotting: one was considered positive by immunoblotting and negative by IFA. 68% of patients with KS and 12% of HIV-1 infected patients without KS were reactive by indirect immunofluorescence assay (IFA). Thus using two different assays, antibodies to KSHV lytic cycle antigens were found 6 to 9 times more frequently among patients with KS than among HIV-1 infected patients without KS. Stated another way, among individuals who were seropositive to KSHV p40 32/35 (91%) had KS. Among those seropositive by the immunofluorescence assay 32/37 (86%) had KS. Thus infection with KSHV, as defined by these serologic markers, carries a high risk of development of KS.
 The recent discovery of genetic sequences representative of a new human herpes virus in KS tumor tissue, taken together with past epidemiologic observations, strongly implicate this novel agent in the pathogenesis of KS. However, these observations, by themselves, do not permit the construction of a unified theory of pathogenesis that accounts for the many mysterious features of KS. For example, the relative contribution of HIV-1, other forms of immunosuppression, geographic factors, sex differences, the role of cytokines and growth factors, and the occurrence of distinct clinical variants must all be eventually understood. By identifying the infection rate in different populations a serologic marker for infection with KSHV would be great aid in unraveling the significance of the new virus in this complicated puzzle.
 One possibility is that KSHV, the putative etiologic agent is, like all the other human herpes viruses, a ubiquitous, or at least widespread virus which infects large segments of the human population. Individuals who are immunosuppressed would have a greater likelihood of developing disease, whereas immunocompetent individuals would remain healthy. This pathogenetic model is similar to that postulated for the role that EBV plays in non-Hodgkin's lymphoma or cytomegalovirus in retinitis in patients with AIDS. If this model is correct a very high proportion of the adult human population might be found to be seropositive for KSHV. The model of a ubiquitous virus selectively causing disease in immunodeficient individuals does not account for classical KS affecting patients who are not immunocompromised nor does it account for the observations that endemic KS in Africa preceded the HIV-1 epidemic. Since many African patients with KS are HIV-1 negative other co-factors must be implicated.
 The other possibility is that KSHV infection occurs selectively in the human population. Transmission may be promoted by sexual behavior that also carries a high risk of acquiring HIV-1. In this scenario seroprevalence of KSHV would be expected to be higher in HIV-1 seropositive and HIV-1 seronegative homosexual men than in other populations. If the virus alone were capable of inducing disease, acquisition of KSHV infection, as monitored by the presence of antibody, would be associated with a high rate of clinically evident KS. However, if KSHV infection needed to accompanied by other co-factors to cause disease, the prevalence of antibody of KSHV might be similar among patients with and without KS. The other co-factors would not be identified in a serologic test for antibodies to KSHV antigens.
 The findings, using tests for antibodies to KSHV lytic cycle antigens, are consistent with the general model in which infection with KSHV is infrequent but associated with a high rate of apparent disease. Only a few HIV-1 infected patients without KS had antibodies to the KSHV lytic cycle antigens; by contrast a very high proportion of HIV-1 infected men who had clinically evident KS were seropositive. This finding suggests that a high proportion of individuals who are dually infected with HIV-1 and KSHV develop KS. However, another interpretation of the data is possible, though this interpretation is novel and no other examples are known among the human herpes virus family. Infection with KSHV might be ubiquitous, antibodies to the virus would not normally be detected in healthy infected individuals. Antibodies would only appear after the virus has been reactivated from the latent into the lytic cycle as might occur during the course of immunosuppression. Thus the two serologic tests that are described would indicate reactivated infection but would not be an index of past exposure to the virus. If this interpretation is correct, it should be possible to demonstrate KSHV DNA sequences or tot isolate the virus from healthy individuals who are KSHV seronegative.
 Regardless of which of these two interpretations is correct, the serologic studies provide a strong correlation between the presence of antibodies to KSHV lytic cycle gene products and clinical KS. Nonetheless there are two groups of patients whose serologic results require further explanation. One group consists of the few patients with positive serology for KSHV p40 without clinical KS. They may have subclinical or visceral disease, or they may develop KS in the future. The other group is the approximately 30% of patients with KS whose sera lacked antibody to p40. The patients with KS who were p40 seronegative were not misclassified since the diagnosis was confirmed in all of them by biopsy (FIG. 25). It is possible that the antibodies being measured are variable and wax and wane with time following infection. The appearance of antibody to p40 may reflect the extent of lytic viral replication which may vary during different phases of the disease. To determine whether this is true prospective studies including serial bleedings are required.
 p40 is likely to be only one among a number of KSHV antigens recognized by the infected patients. Antibody recognition of other KSHV antigens may not be possible on immunoblots because they comigrate with EBV polypeptides, because the BCBL-1 cells cannot be induced to express these antigens, or because the antigens are of low abundance or denatured on the immunoblots. In some individuals serum antibodies to p40 may be consumed in immune complexes with p40 antigen in the circulation. Thus detection of p40 on immunoblots may not be of optimal sensitivity. In this connection three sera recognized antigens in immunofluorescence tests but did not react with p40 on western blots. The serologic test employing whole BCBL-1 cells as antigen are clearly first generation assays to be improved by better characterization of the KSHV gene products and preparation of recombinant antigens.
 Lack of a serologic response to p40 could also reflect severely impaired humoral immunity. Although humoral immunity is usually relatively intact in HIV infection, examples of impaired antibody response have been described. For instance, some individuals are known to have impaired antibody responses to parvovirus B19(40 and others have been observed to lose antibodies to hepatitis B surface antigen (41]. An association between the degree of immunosuppression, as monitored by the number of CD4 cells, and the presence or absence of antibody p40 among patients with KS was not found (FIG. 25). Furthermore all the patients with or without antibodies to KSHV p40 had antibodies to EBV p21 suggesting an intact humoral immune response.
 In these serologic studies, as in the genetic probe studies previously reported, KSHV infection was found in the majority, but not all, patients with KS. Assuming that methodologic explanations do not account exclusively for the seronegative patients, other pathways, in addition to infection with KSHV, may lead to development of KS. In fact, most data suggest that the pathogenesis of KS is a multifactorial process. It has been observed that the product of the HIV-tat gene stimulates growth of KS tissue culture cells  and can induce KS-like lesions in mice . These findings suggest a direct role for HIV-1 in the pathogenesis of KS, at least in HIV-infected hosts. In other settings, other growth factors may play a similar or complementary function. Interleukin-6 and basic fibroblast growth factor are both known stimulate growth of KS cells in vitro . Interleukin-6 is also produced in AIDS-KS derived cell culture . Thus, KS pathogenesis may involve autocrine and paracrine growth factors together with infection with KSHV in some patients or with certain strains of HIV-1 in other patients. If infection with KSHV is the sine qua non of this process on would expect to see evidence of KSHV infection in all patients with KS.
 In summary, an immunoblotting and a immunofluorescence screening assay for detection of antibodies to lytic cycle antigens of KSHV is disclosed. These assays should permit detailed seroepidemiologic investigations of KSHV. The findings support the notion of a strong association between infection with KSHV and the development of KS in HIV-infected patients. Infection with KSHV, as defined by these serologic assays, appears to carry an extremely high risk of development of clinical KS.
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tctataatct taaagttggt 5100ataagactgg tcgctcgtta tggccagccg gcactccggt agtatctgcg tgtcctcgaa 5160ttcgtggccg cgtacgactg gcttggagtg caggtaaacg ccaagagatg cggtctcttc 5220gcctacgcac aagtggcttc ttaacgcgta ggggtgcggt gagagcatga tccgtagcaa 5280cgatagttcc gggtgcctag ccgcgtagag tggcagggta gacgagtccg gagtcccaaa 5340cttttcgaac aacagtggca tcgggacttc aggattagag actcccacca tggccgccac 5400cgccggagag gtcaagacgt gaaacacgcg ctcgcctgtc gacaggcgcg ccgcgccctc 5460tactagacta gccttcacgt ccggaactcg taacatagct tagaccagcg gacggacgca 5520acgtacgcgg ggatcggctg gcggtgtctg ctcgttggac gcggccgttc ggtggcgcca 5580gtgcaggcct agtttgcgaa tggcgtgacg gacaatttgt ggctttagag cggcgaaccg 5640atgacccgtg gtggcgacga acgaaatgaa gtttgcattg cggcccaact cgtctagcct 5700ggtcttcttg tttcgggcat agattttcgg gattaggtta cactttttat atcccagtac 5760tgcgcactcg tgtttgcttt tagtgtgact gattatcttc tttgagaagt caaacaggcc 5820ccgggcggcg gctcgcctaa tgcaagccac gtcaagcctg agaaacgaac agcattccac 5880cagacactcc aggaaccttt tgtgtagcgt ctgtatttgg gaacggtttc tgtgctcaag 5940tagggagaat attctatttt tgtttccgtc gatgcgcgcg tgctggtccg tgagaatggg 6000cgccagctcg tggcgaatct gttccacaag aggctgcccg tacactttag aaatcgtggc 6060tgtcgcggcc ttaaaccagg acacgtttag cccatccttg ctggagacca cagatggaaa 6120gtttgtggtc caaaatacgt tttttcgccc cattctcacc atgtactggt tttccagtcc 6180gtgcaggtcc aacgtggagt tccaatttgc tatcgataca ggaaatatgt gcctgattgg 6240cagaaagcat ttcagcgtac ccattgcgaa gagaaagtgc agcatgtccc cactgatgtt 6300gatgtttatt gcggtgcctt gacacatgtt gtcggaaaaa aacacgctta tggtaaaaga 6360aggttccttt acggagtact ttcgtataac aaaattgttg gtcaatctgg ggatgtttaa 6420aatagtcttt tgcagggtgt taggaacgtg gcagcttatc ttagtgttaa tcaccatgtt 6480ggtgttgaat atggtgatct tgaagttttc caaactgacg tgttttgtgg gttccagcat 6540gtctgacact gtagagctgc ccagagtccg cgcgtccgtg gccgcgtatc gttggaagca 6600cgcctgcaaa tttcctttca tggctgctcg ccggtctttc ggcgcgtacc ggattcttga 6660aagcgtcgcc gccaggagac gcggtgtctc gtgggtgcct aaaaagtttg cgcaggggtg 6720cagtccgctg cacgagtggc cgatgcagtc tgccactgcc atacacatga cgagtctgta 6780gatggccggt gtgcccggat acactagata gtaggtacaa tctggggtac tgacgaccac 6840cctgtatggc tttggtccgg ggtccttgcg ttggattttt acgtgcagac gggacacgag 6900ctggtttaga gccagctgaa agcccaccag atcccgtccg ttaaccttga cgtcctggtg 6960cttactctgt ttcgacaggt tcttcagcac ggtgggcagt cgctctacgt tgtgagcgat 7020ggcacggcgc agcgagacca gctctccgtg ccacccccac gtggccatga agctgctgat 7080gttaaacttt aaaaaatgta gctgtgcgtc tggggatgcg ggtggcatta ttgaaaacga 7140gagatgcttc aggctctcca ggagtgcaaa ataattttga tagattgtgg gttgtagact 7200atggggcaac accgccagaa acgcatgaaa acactgttcg aactcccaga actccaggta 7260cctgcacact atcctgaaca tggctttgta acatatggtg cacgttagta gcgcgggaag 7320atacagcgag cgtagctccc tgaattcgca gggtttatca caatcatcgg taagttccca 7380tgatcccacc gcaggtaggt agttgtcggt gtctatctgt ccgcgcgtaa acactccacc 7440accgtcaatt attaaacctt cgccgctgta ccgtcgaccc acttttccca aaagagtccc 7500ttcttgatgt ataaaagggt ggaggcgttc ccccaggagt agtctgcgta tcgctctgca 7560ggcgaaaaag gtgggctcgg gctgcatcat cttatcaaga ccttctaagg tcagctctgc 7620ctgcaggtgc gagttggtgg ccagacagca gaatatttcc agctgtgatt cccaagtcgc 7680ttgataacac gtggtctgcg gactcgtcgt cagggaggcg ctcggtggca gtagtagggg 7740gccctcgagc gctgccatgg aggcgacctt ggagcaacga cctttcccgt acctcgccac 7800ggaggccaac ctcctaacgc agattaagga gtcggctgcc gacggactct tcaagagctt 7860tcagctattg ctcggcaagg acgccagaga aggcagtgtc cgtttcgaag cgctactggg 7920cgtatatacc aatgtggtgg agtttgttaa gtttctggag accgccctcg ccgccgcttg 7980cgtcaatacc gagttcaagg acctgcggag aatgatagat ggaaaaatac agtttaaaat 8040ttcaatgccc actattgccc acggagacgg gaggaggccc aacaagcaga gacagtatat 8100cgtcatgaag gcttgcaata agcaccacat cggtgcggag attgagcttg cggccgcaga 8160catcgagctt ctcttcgccg agaaagagac gcccttggac ttcacagagt acgcgggtgc 8220catcaagacg attacgtcgg ctttgcagtt tggtatggac gccctagaac gggggctagt 8280ggacacggtt ctcgcagtta aacttcggca cgctccaccc gtctttattt taaagacgct 8340gggcgatccc gtctactctg agaggggcct caaaaaggcc gtcaagtctg acatggtatc 8400catgttcaag gcacacctca tagaacattc attttttcta gataaggccg agctcatgac 8460aagggggaag cagtatgtcc taaccatgct ctccgacatg ctggccgcgg tgtgcgagga 8520taccgtcttt aagggtgtca gcacgtacac cacggcctct gggcagcagg tggccggcgt 8580cctggagacg acggacagcg tcatgagacg gctgatgaac ctgctggggc aagtggaaag 8640tgccatgtcc gggcccgcgg cctacgccag ctacgttgtc aggggtgcca acctcgtcac 8700cgccgttagc tacggaaggg cgatgagaaa ctttgaacag tttatggcac gcatagtgga 8760ccatcccaac gctctgccgt ctgtggaagg tgacaaggcc gctctggcgg acggacacga 8820cgagattcag agaacccgca tcgccgcctc tctcgtcaag ataggggata agtttgtggc 8880cattgaaagt ttgcagcgca tgtacaacga gactcagttt ccctgcccac tgaaccggcg 8940catccagtac acctatttct tccctgttgg ccttcacctt cccgtgcccc gctactcgac 9000atccgtctca gtcaggggcg tagaatcccc ggccatccag tcgaccgaga cgtgggtggt 9060taataaaaac aacgtgcctc tttgcttcgg ttaccaaaac gccctcaaaa gcatatgcca 9120ccctcgaatg cacaacccca cccagtcagc ccaggcacta aaccaagctt ttcccgatcc 9180cgacggggga catgggtacg gtctcaggta tgagcagacg ccaaacatga acctattcag 9240aacgttccac cagtattaca tggggaaaaa cgtggcattt gttcccgatg tggcccaaaa 9300agcgctcgta accacggagg atctactgca cccaacctct caccgtctcc tcagattgga 9360ggtccacccc ttctttgatt tttttgtgca cccctgtcct ggagcgagag gatcgtaccg 9420cgccacccac agaacaatgg ttggaaatat accacaaccg ctcgctccaa gggagtttca 9480ggaaagtaga ggggcgcagt tcgacgctgt gacgaatatg acacacgtca tagaccagct 9540aactattgac gtcatacagg agacggcatt tgaccccgcg tatcccctgt tctgctatgt 9600aatcgaagca atgattcacg gacaggaaga aaaattcgtg atgaacatgc ccctcattgc 9660cctggtcatt caaacctact gggtcaactc gggaaaactg gcgtttgtga acagttatca 9720catggttaga ttcatctgta cgcatattgg gaatggaagc atccctaagg aggcgcacgg 9780ccactaccgg aaaatcttag gcgagctcat cgcccttgag caggcgcttc tcaagctcgc 9840gggacacgag acggtgggtc ggacgccgat cacacatctg gtttcggctc tcctcgaccc 9900gcatctgctg cctccctttg cctaccacga tgtctttacg gatcttatgc agaagtcatc 9960cagacaaccc ataatcaaga tcggggatca aaactacgac aaccctcaaa atagggcgac 10020attcatcaac ctcaggggtc gcatggagga cctagtcaat aaccttgtta acatttacca 10080gacaagggtc aatgaggacc atgacgagag acacgtcctg gacgtggcgc ccctggacga 10140gaatgactac aacccggtcc tcgagaagct attctactat gttttaatgc cggtgtgcag 10200taacggccac atgtgcggta tgggggtcga ctatcaaaac gtggccctga cgctgactta 10260caacggcccc gtctttgcgg acgtcgtgaa cgcacaggat gatattctac tgcacctgga 10320gaacggaacc ttgaaggaca ttctgcaggc aggcgacata cgcccgacgg tggacatgat 10380cagggtgctg tgcacctcgt ttctgacgtg ccctttcgtc acccaggccg ctcgcgtgat 10440cacaaagcgg gacccggccc agagttttgc cacgcacgaa tacgggaagg atgtggcgca 10500gaccgtgctt gttaatggct ttggtgcgtt cgcggtggcg gaccgctctc gcgaggcggc 10560ggagactatg ttttatccgg taccctttaa caagctctac gctgacccgt tggtggctgc 10620cacactgcat ccgctcctgc caaactatgt caccaggctc cccaaccaga gaaacgcggt 10680ggtctttaac gtgccatcca atctcatggc agaatatgag gaatggcaca agtcgcccgt 10740cgcggcgtat gccgcgtctt gtcaggccac cccgggcgcc attagcgcca tggtgagcat 10800gcaccaaaaa ctatctgccc ccagtttcat ttgccaggca aaacaccgca tgcaccctgg 10860ttttgccatg acagtcgtca ggacggacga ggttctagca gagcacatcc tatactgctc 10920cagggcgtcg acatccatgt ttgtgggctt gccttcggtg gtacggcgcg aggtacgttc 10980ggacgcggtg acttttgaaa ttacccacga gatcgcttcc ctgcacaccg cacttggcta 11040ctcatcagtc atcgccccgg cccacgtggc cgccataact acagacatgg gagtacattg 11100tcaggacctc tttatgattt tcccagggga cgcgtatcag gaccgccagc tgcatgacta 11160tatcaaaatg aaagcgggcg tgcaaaccgg ctcaccggga aacagaatgg atcacgtggg 11220atacactgct ggggttcctc gctgcgagaa cctgcccggt ttgagtcatg gtcagctggc 11280aacctgcgag ataattccca cgccggtcac atctgacgtt gcctatttcc agacccccag 11340caacccccgg gggcgtgcgg cgtcggtcgt gtcgtgtgat gcttacagta acgaaagcgc 11400agagcgtttg ttctacgacc attcaatacc agaccccgcg tacgaatgcc ggtccaccaa 11460caacccgtgg gcttcgcagc gtggctccct cggcgacgtg ctatacaata tcacctttcg 11520ccagactgcg ctgccgggca tgtacagtcc ttgtcggcag ttcttccaca aggaagacat 11580tatgcggtac aatagggggt tgtacacttt ggttaatgag tattctgcca ggcttgctgg 11640ggcccccgcc accagcacta cagacctcca gtacgtcgtg gtcaacggta cagacgtgtt 11700tttggaccag ccttgccata tgctgcagga ggcctatccc acgctcgccg ccagccacag 11760agttatgctt gccgagtaca tgtcaaacaa gcagacacac gccccagtac acatgggcca 11820gtatctcatt gaagaggtgg cgccgatgaa gagactatta aagctcggaa acaaggtggt 11880gtattagcta acccttctag cgttggctag tcatggcact cgacaagagt atagtggtta 11940acttcacctc cagactcttc gctgatgaac tggccgccct tcagtcaaaa atagggagcg 12000tactgccgct cggagattgc caccgtttac aaaatataca ggcattgggc ctggggtgcg 12060tatgctcacg tgagacatct ccggactaca tccaaattat gcagtatcta tccaagtgca 12120cactcgctgt cctggaggag gttcgcccgg acagcctgcg cctaacgcgg atggatccct 12180ctgacaacct tcagataaaa aacgtatatg cccccttttt tcagtgggac agcaacaccc 12240agctagcagt gctaccccca ttttttagcc gaaaggattc caccattgtg ctcgaatcca 12300acggatttga ccccgtgttc cccatggtcg tgccgcagca actggggcac gctattctgc 12360agcagctgtt ggtgtaccac atctactcca aaatatcggc cggggccccg gatgatgtaa 12420atatggcgga acttgatcta tataccacca atgtgtcatt tatggggcgc acatatcgtc 12480tggacgtaga caacacggat ccacgtactg ccctgcgagt gcttgacgat ctgtccatgt 12540acctttgtat cctatcagcc ttggttccca gggggtgtct ccgtctgctc acggcgctcg 12600tgcggcacga caggcatcct ctgacagagg tgtttgaggg ggtggtgcca gatgaggtga 12660ccaggataga tctcgaccag ttgagcgtcc cagatgacat caccaggatg cgcgtcatgt 12720tctcctatct tcagagtctc agttctatat ttaatcttgg ccccagactg cacgtgtatg 12780cctactcggc agagactttg gcggcctcct gttggtattc cccacgctaa cgatttgaag 12840cggggggggt atggcgtcat ctgatattct gtcggttgca aggacggatg acggctccgt 12900ctgtgaagtc tccctgcgtg gaggtaggaa aaaaactacc gtctacctgc cggacactga 12960accctgggtg gtagagaccg acgccatcaa agacgccttc ctcagcgacg ggatcgtgga 13020tatggctcga aagcttcatc gtggtgccct gccctcaaat tctcacaacg gcttgaggat 13080ggtgcttttt tgttattgtt acttgcaaaa ttgtgtgtac ctagccctgt ttctgtgccc 13140ccttaatcct tacttggtaa ctccctcaag cattgagttt gccgagcccg ttgtggcacc 13200tgaggtgctc ttcccacacc cggctgagat gtctcgcggt tgcgatgacg cgattttctg 13260taaactgccc tataccgtgc ctataatcaa caccacgttt ggacgcattt acccgaactc 13320tacacgcgag ccggacggca ggcctacgga ttactccatg gcccttagaa gggcttttgc 13380agttatggtt aacacgtcat gtgcaggagt gacattgtgc cgcggagaaa ctcagaccgc 13440atcccgtaac cacactgagt gggaaaatct gctggctatg ttttctgtga ttatctatgc 13500cttagatcac aactgtcacc cggaagcact gtctatcgcg agcggcatct ttgacgagcg 13560tgactatgga ttattcatct ctcagccccg gagcgtgccc tcgcctaccc cttgcgacgt 13620gtcgtgggaa gatatctaca acgggactta cctagctcgg cctggaaact gtgacccctg 13680gcccaatcta tccacccctc ccttgattct aaattttaaa taaaggtgtg tcactggtta 13740caccacgatt aaaaaccact cactgagatg tctttttaac cgctaaggga ttataccggg 13800atttaaaacc gcccactgat ttttttacgc taagagttgg gtgcttgggg ggttttgcat 13860tgctctgttg taaactatat ataagttaaa ccaaaattcg cagggagaca aggtgacggt 13920ggtgagaact cagttgagag tcagagaata cagtgctaat cagggtagat gagcatgact 13980ttccccgtct ccagtcaccg gaggaatggt ggacggctcc gtcctggtgc gaatggccac 14040caagcctccc gtgattggtc ttataacagt gctcttcctc ctagtcatag gcgcctgcgt 14100ctactgctgc attcgcgtgt tcctggcggc tcgactgtgg cgcgccaccc cactaggcag 14160ggccaccgtg gcgtatcagg tccttcgcac cctgggaccg caggccgggt cacatgcacc 14220gccgacggtg ggcatagcta cccaggagcc ctaccgtaca atatacatgc cagattagaa 14280cggggtgtgt gctataatgg atggctatgg gggggggctg tagataattg agcgctgtgc 14340ttttattgtg gggatatggg cttgtacatg tgtctatcat cggtagccat aaaatgggcc 14400atgacaactg ccacaagtaa gtcgtccgac atgtgctttt gcttggcgct gtatgactgc 14460cctccatccc taagcgggac gcacttgatc gcgcggacct gttctaccag gtaggtcacc 14520gggtcaaatg atattttgat ggtgttggac accaccgtct ggctggcgct cagggtgccg 14580gagttcagag cgtagatgaa tgtctcaaac gcggaggatt tctcgcctcc caacatgtaa 14640attggccact gcagggcgct gctcttgtca gtatagtgta gaaaatgtat ggggagcggg 14700catatttcgt taaggacggt tgcaatggcc accccagaat cttggctgct gttgccttcg 14760accgccgcgt tcacgcgctc aattgtggtg tggagcacag cgatcgcctt aatcatcgtg 14820catgcgcagg acgctatctc gtaagcagct gcgccagtga ggtcgcgcag gaagaaatgc 14880tccatgccca atatgaggct tctggtggga gtctgagtac tcgtgacaac ggcgcccacg 14940ccagtaccgg acgcctccgt gttgttcgta tacgcggggt cgatgtaaac aaacagctgt 15000tttccaaggc acttctgaac
ctcctgggcg gtggtgtcta cccgacacat gtcaaactgt 15060gtcagcgctg cgtcacccac cacgcggtaa agcgtagcat ttgacgacgc tgctccctcg 15120cccattagtt cggtgtcgaa tgccccctcc ataaagaggt tggtggtggt tttgatggat 15180tcgtcgatgg tgatgtacgt cggaatgtgc agtctgtaac aaggacagga cactagtgcg 15240tcttgcaggt ggaaatcttc tcggtggtcc gcacacacgt aactgaccac attcagcatc 15300ttttcctggg cgttcctgag gttaagcagg aaactcgtgg agcggtctga cgagttcacg 15360gatgatataa atataagctt ggcgtctttc tgaagcatga aacccagaat agccggcagt 15420gcatcctttt taataaaatt cgcctcgtct acgtagagca ggttaaaggt ctgtccccga 15480atgctctgca gacacggaaa gacacaaaag aggggctcat aagcggctaa cagtaaagga 15540gaggaggcga acagtgcgtg gctcttggtt cttgggaata aaagggggcg tgtgtgccga 15600tcgatcgtat gggtgagcca gtggatcctg gacatgtggt gaatgagaaa gattttgagg 15660agtgtgaaca atttttcagt caacccctta gggagcaagt ggtcgcgggg gtcagggcac 15720tcgacggcct cggtctcgct gactctctat gtcacaaaac agaaagactc tgcctgctga 15780tggacctggt gggcacggag tgctttgcga gggtgtgccg cctagacacc ggtgcgaaat 15840gaagagtgtg gcgagtccct tatgtcagtt ccacggcgtg ttttgcctgt accagtgtcg 15900ccagtgcctg gcataccacg tgtgtgatgg gggcgccgaa tgcgttctcc tgcatacgcc 15960ggagagcgtc atctgcgaac taacgggtaa ctgcatgctc ggcaacattc aagagggcca 16020gtttttaggg ccggtaccgt atcggacttt ggataaccag gttgacaggg acgcatatca 16080cgggatgcta gcgtgtctga aacgggacat tgtgcggtat ttgcagacat ggccggacac 16140caccgtaatc gtgcaggaaa tagccctggg ggacggcgtc accgacacca tctcggccat 16200tatagatgaa acattcggtg agtgtcttcc cgtactgggg gaggcccaag gcgggtacgc 16260cctggtctgt agcatgtatc tgcacgttat cgtctccatc tattcgacaa aaacggtgta 16320caacagtatg ctatttaaat gcacaaagaa taaaaagtac gactgcattg ccaagcgggt 16380gcggacaaaa tggatgcgca tgctatcaac gaaagatacg taggtcctcg ctgccaccgt 16440ttggcccacg tggtgctgcc taggaccttt ctgctgcatc acgccatacc cctggagccc 16500gagatcatct tttccaccta cacccggttc agccggtcgc cagggtcatc ccgccggttg 16560gtggtgtgtg ggaaacgtgt cctgccaggg gaggaaaacc aacttgcgtc ttcaccttct 16620ggtttggcgc ttagcctgcc tctgttttcc cacgatggga actttcatcc atttgacatc 16680tcggtactgc gcatttcctg ccctggttct aatcttagtc ttactgtcag atttctctat 16740ctatctctgg tggtggctat gggggcggga cggaataatg cgcggagtcc gaccgttgac 16800ggggtatcgc cgccagaggg cgccgtagcc caccctttgg aggaactgca gaggctggcg 16860cgtgctacgc cggacccggc actcacccgt ggaccgttgc aggtcctgac cggccttctc 16920cgcgcagggt cagacggaga ccgcgccact caccacatgg cgctcgaggc tccgggaacc 16980gtgcgtggag aaagcctaga cccgcctgtt tcacagaagg ggccagcgcg cacacgccac 17040aggccacccc ccgtgcgact gagcttcaac cccgtcaatg ccgatgtacc cgctacctgg 17100cgagacgcca ctaacgtgta ctcgggtgct ccctactatg tgtgtgttta cgaacgcggt 17160ggccgtcagg aagacgactg gctgccgata ccactgagct tcccagaaga gcccgtgccc 17220ccgccaccgg gcttagtgtt catggacgac ttgttcatta acacgaagca gtgcgacttt 17280gtggacacgc tagaggccgc ctgtcgcacg caaggctaca cgttgagaca gcgcgtgcct 17340gtcgccattc ctcgcgacgc ggaaatcgca gacgcagtta aatcgcactt tttagaggcg 17400tgcctagtgt tacgggggct ggcttcggag gctagtgcct ggataagagc tgccacgtcc 17460ccgccccttg gccgccacgc ctgctggatg gacgtgttag gattatggga aagccgcccc 17520cacactctag gtttggagtt acgcggcgta aactgtggcg gcacggacgg tgactggtta 17580gagattttaa aacagcccga tgtgcaaaag acagtcagcg ggagtcttgt ggcatgcgtg 17640atcgtcacac ccgcattgga agcctggctt gtgttacctg ggggttttgc tattaaagcc 17700cgctataggg cgtcgaagga ggatctggtg ttcattcgag gccgctatgg ctagccggag 17760gcgcaaactt cggaatttcc taaacaagga atgcatatgg actgttaacc caatgtcagg 17820ggaccatatc aaggtcttta acgcctgcac ctctatctcg ccggtgtatg accctgagct 17880ggtaaccagc tacgcactga gcgtgcctgc ttacaatgtg tctgtggcta tcttgctgca 17940taaagtcatg ggaccgtgtg tggctgtggg aattaacgga gaaatgatca tgtacgtcgt 18000aagccagtgt gtttctgtgc ggcccgtccc ggggcgcgat ggtatggcgc tcatctactt 18060tggacagttt ctggaggaag catccggact gagatttccc tacattgctc cgccgccgtc 18120gcgcgaacac gtacctgacc tgaccagaca agaattagtt catacctccc aggtggtgcg 18180ccgcggcgac ctgaccaatt gcactatggg tctcgaattc aggaatgtga acccttttgt 18240ttggctcggg ggcggatcgg tgtggctgct gttcttgggc gtggactaca tggcgttctg 18300tccgggtgtc gacggaatgc cgtcgttggc aagagtggcc gccctgctta ccaggtgcga 18360ccacccagac tgtgtccact gccatggact ccgtggacac gttaatgtat ttcgtgggta 18420ctgttctgcg cagtcgccgg gtctatctaa catctgtccc tgtatcaaat catgtgggac 18480cgggaatgga gtgactaggg tcactggaaa cagaaatttt ctgggtcttc tgttcgatcc 18540cattgtccag agcagggtaa cagctctgaa gataactagc cacccaaccc ccacgcacgt 18600cgagaatgtg ctaacaggag tgctcgacga cggcaccttg gtgccgtccg tccaaggcac 18660cctgggtcct cttacgaatg tctgactact tcagccgctt gctgatatat gagtgtaaaa 18720aacttaaggc cctgggctta cgttcttatt gaagcatgtt gcgcacatca gcgagctgga 18780ccgtcctccg ggtcgcgtgt agattatggt tccgttctcc ttcttgatgt ttaaattttt 18840gggggggaac caccgacaaa gcgtctttat gatttccgcg aacacggagt tggctacgtg 18900cttttggtgg gctacgtacc caatgttaat gttctctacg gatgccagta gcatgctgat 18960gatcgccacc actatccatg tctttccgtg tctccttggt attaggaata cgcttgcctt 19020ttgcttaaac gtctgtaaaa cactgtttgg agtttcaaat aaaccgaagt actgcttaaa 19080caatccaaac aactggtgcg tcttttgtgg ggccttgatt gaaaccaaaa agaaaaaagt 19140gtgcattact agctgctgtt ggaagggctc cagccagtgc accccgggaa cgtaacagcc 19200gttcagaaag gacgaaaggt taaccagaaa agcctgaagt tcgcggtaga cagagcaggc 19260gtgcagggag tcgtgtgttt ttctgcccgc ctggtactcg accagttgat cggccgtgga 19320gacgtgcgcg tcctcgcgca cacaccgcat ctgcaagtat gttgataggg actccaatag 19380gcgcggcttt gcggggacgt tgtcctcgga cggtctgggg gttcccacgt cgggatttgc 19440tgacgtgggc gtggcgggat ggtgccgtgt gcagtatgtt tccaggaccg aactgtatga 19500gtttattctg tgcaccacgc caataaaagg gtgcgccatc cgtgccgttt tgggacagtg 19560tcgcgtgaat gtcggggcac tcagttccca cctctctccg gcgtctttgg cggtctcctc 19620caggttggcg gcaaggcgct ccctgtgacg gctgagcagc atgtttgctt tgagctcgct 19680cgtgtccgag ggtgacccgg aggtgaccag taggtacgtc aagggcgtac aacttgccct 19740ggaccttagc gagaacacac ctggacaatt taagttgata gaaactcccc tgaacagctt 19800cctcttggtt tccaacgtga tgcccgaggt ccagccaatc tgcagtggcc ggccggcctt 19860gcggccagac tttagtaatc tccacttgcc tagactggag aagctccaga gagtcctcgg 19920gcagggtttc ggggcggcgg gtgaggaaat cgcactggac ccgtctcacg tagaaacaca 19980cgaaaagggc caggtgttct acaaccacta tgctaccgag gagtggacgt gggctttgac 20040tctgaataag gatgcgctcc ttcgggaggc tgtagatggc ctgtgtgacc ccggaacttg 20100gaagggtctt cttcctgacg acccccttcc gttgctatgg ctgctgttca acggacccgc 20160ctctttttgt cgggccgact gttgcctgta caagcagcac tgcggttacc cgggcccggt 20220gctacttcca ggtcacatgt acgctcccaa acgggatctt ttgtcgttcg ttaatcatgc 20280cctgaagtac accaagtttc tatacggaga tttttccggg acatgggcgg cggcttgccg 20340cccgccattc gctacttctc ggatacaaag ggtagtgagt cagatgaaaa tcatagatgc 20400ttccgacact tacatttccc acacctgcct cttgtgtcac atatatcagc aaaatagcat 20460aattgcgggt caggggaccc acgtgggtgg aatcctactg ttgagtggaa aagggaccca 20520gtatataaca ggcaatgttc agacccaaag gtgtccaact acgggcgact atctaatcat 20580cccatcgtat gacataccgg cgatcatcac catgatcaag gagaatggac tcaaccaact 20640ctaaaagaga gtttattaag tcggctctgg aggccaacat caacaggagg gcagctgtat 20700cgctatttga 2071024128DNAKaposi's sarcoma-associated herpesvirus 2atggaggcga ccttggagca acgacctttc ccgtacctcg ccacggaggc caacctccta 60acgcagatta aggagtcggc tgccgacgga ctcttcaaga gctttcagct attgctcggc 120aaggacgcca gagaaggcag tgtccgtttc gaagcgctac tgggcgtata taccaatgtg 180gtggagtttg ttaagtttct ggagaccgcc ctcgccgccg cttgcgtcaa taccgagttc 240aaggacctgc ggagaatgat agatggaaaa atacagttta aaatttcaat gcccactatt 300gcccacggag acgggaggag gcccaacaag cagagacagt atatcgtcat gaaggcttgc 360aataagcacc acatcggtgc ggagattgag cttgcggccg cagacatcga gcttctcttc 420gccgagaaag agacgccctt ggacttcaca gagtacgcgg gtgccatcaa gacgattacg 480tcggctttgc agtttggtat ggacgcccta gaacgggggc tagtggacac ggttctcgca 540gttaaacttc ggcacgctcc acccgtcttt attttaaaga cgctgggcga tcccgtctac 600tctgagaggg gcctcaaaaa ggccgtcaag tctgacatgg tatccatgtt caaggcacac 660ctcatagaac attcattttt tctagataag gccgagctca tgacaagggg gaagcagtat 720gtcctaacca tgctctccga catgctggcc gcggtgtgcg aggataccgt ctttaagggt 780gtcagcacgt acaccacggc ctctgggcag caggtggccg gcgtcctgga gacgacggac 840agcgtcatga gacggctgat gaacctgctg gggcaagtgg aaagtgccat gtccgggccc 900gcggcctacg ccagctacgt tgtcaggggt gccaacctcg tcaccgccgt tagctacgga 960agggcgatga gaaactttga acagtttatg gcacgcatag tggaccatcc caacgctctg 1020ccgtctgtgg aaggtgacaa ggccgctctg gcggacggac acgacgagat tcagagaacc 1080cgcatcgccg cctctctcgt caagataggg gataagtttg tggccattga aagtttgcag 1140cgcatgtaca acgagactca gtttccctgc ccactgaacc ggcgcatcca gtacacctat 1200ttcttccctg ttggccttca ccttcccgtg ccccgctact cgacatccgt ctcagtcagg 1260ggcgtagaat ccccggccat ccagtcgacc gagacgtggg tggttaataa aaacaacgtg 1320cctctttgct tcggttacca aaacgccctc aaaagcatat gccaccctcg aatgcacaac 1380cccacccagt cagcccaggc actaaaccaa gcttttcccg atcccgacgg gggacatggg 1440tacggtctca ggtatgagca gacgccaaac atgaacctat tcagaacgtt ccaccagtat 1500tacatgggga aaaacgtggc atttgttccc gatgtggccc aaaaagcgct cgtaaccacg 1560gaggatctac tgcacccaac ctctcaccgt ctcctcagat tggaggtcca ccccttcttt 1620gatttttttg tgcacccctg tcctggagcg agaggatcgt accgcgccac ccacagaaca 1680atggttggaa atataccaca accgctcgct ccaagggagt ttcaggaaag tagaggggcg 1740cagttcgacg ctgtgacgaa tatgacacac gtcatagacc agctaactat tgacgtcata 1800caggagacgg catttgaccc cgcgtatccc ctgttctgct atgtaatcga agcaatgatt 1860cacggacagg aagaaaaatt cgtgatgaac atgcccctca ttgccctggt cattcaaacc 1920tactgggtca actcgggaaa actggcgttt gtgaacagtt atcacatggt tagattcatc 1980tgtacgcata ttgggaatgg aagcatccct aaggaggcgc acggccacta ccggaaaatc 2040ttaggcgagc tcatcgccct tgagcaggcg cttctcaagc tcgcgggaca cgagacggtg 2100ggtcggacgc cgatcacaca tctggtttcg gctctcctcg acccgcatct gctgcctccc 2160tttgcctacc acgatgtctt tacggatctt atgcagaagt catccagaca acccataatc 2220aagatcgggg atcaaaacta cgacaaccct caaaataggg cgacattcat caacctcagg 2280ggtcgcatgg aggacctagt caataacctt gttaacattt accagacaag ggtcaatgag 2340gaccatgacg agagacacgt cctggacgtg gcgcccctgg acgagaatga ctacaacccg 2400gtcctcgaga agctattcta ctatgtttta atgccggtgt gcagtaacgg ccacatgtgc 2460ggtatggggg tcgactatca aaacgtggcc ctgacgctga cttacaacgg ccccgtcttt 2520gcggacgtcg tgaacgcaca ggatgatatt ctactgcacc tggagaacgg aaccttgaag 2580gacattctgc aggcaggcga catacgcccg acggtggaca tgatcagggt gctgtgcacc 2640tcgtttctga cgtgcccttt cgtcacccag gccgctcgcg tgatcacaaa gcgggacccg 2700gcccagagtt ttgccacgca cgaatacggg aaggatgtgg cgcagaccgt gcttgttaat 2760ggctttggtg cgttcgcggt ggcggaccgc tctcgcgagg cggcggagac tatgttttat 2820ccggtaccct ttaacaagct ctacgctgac ccgttggtgg ctgccacact gcatccgctc 2880ctgccaaact atgtcaccag gctccccaac cagagaaacg cggtggtctt taacgtgcca 2940tccaatctca tggcagaata tgaggaatgg cacaagtcgc ccgtcgcggc gtatgccgcg 3000tcttgtcagg ccaccccggg cgccattagc gccatggtga gcatgcacca aaaactatct 3060gcccccagtt tcatttgcca ggcaaaacac cgcatgcacc ctggttttgc catgacagtc 3120gtcaggacgg acgaggttct agcagagcac atcctatact gctccagggc gtcgacatcc 3180atgtttgtgg gcttgccttc ggtggtacgg cgcgaggtac gttcggacgc ggtgactttt 3240gaaattaccc acgagatcgc ttccctgcac accgcacttg gctactcatc agtcatcgcc 3300ccggcccacg tggccgccat aactacagac atgggagtac attgtcagga cctctttatg 3360attttcccag gggacgcgta tcaggaccgc cagctgcatg actatatcaa aatgaaagcg 3420ggcgtgcaaa ccggctcacc gggaaacaga atggatcacg tgggatacac tgctggggtt 3480cctcgctgcg agaacctgcc cggtttgagt catggtcagc tggcaacctg cgagataatt 3540cccacgccgg tcacatctga cgttgcctat ttccagaccc ccagcaaccc ccgggggcgt 3600gcggcgtcgg tcgtgtcgtg tgatgcttac agtaacgaaa gcgcagagcg tttgttctac 3660gaccattcaa taccagaccc cgcgtacgaa tgccggtcca ccaacaaccc gtgggcttcg 3720cagcgtggct ccctcggcga cgtgctatac aatatcacct ttcgccagac tgcgctgccg 3780ggcatgtaca gtccttgtcg gcagttcttc cacaaggaag acattatgcg gtacaatagg 3840gggttgtaca ctttggttaa tgagtattct gccaggcttg ctggggcccc cgccaccagc 3900actacagacc tccagtacgt cgtggtcaac ggtacagacg tgtttttgga ccagccttgc 3960catatgctgc aggaggccta tcccacgctc gccgccagcc acagagttat gcttgccgag 4020tacatgtcaa acaagcagac acacgcccca gtacacatgg gccagtatct cattgaagag 4080gtggcgccga tgaagagact attaaagctc ggaaacaagg tggtgtat 412831376PRTKaposi's sarcoma-associated herpesvirus 3Met Glu Ala Thr Leu Glu Gln Arg Pro Phe Pro Tyr Leu Ala Thr Glu1 5 10 15Ala Asn Leu Leu Thr Gln Ile Lys Glu Ser Ala Ala Asp Gly Leu Phe 20 25 30Lys Ser Phe Gln Leu Leu Leu Gly Lys Asp Ala Arg Glu Gly Ser Val 35 40 45Arg Phe Glu Ala Leu Leu Gly Val Tyr Thr Asn Val Val Glu Phe Val 50 55 60Lys Phe Leu Glu Thr Ala Leu Ala Ala Ala Cys Val Asn Thr Glu Phe65 70 75 80Lys Asp Leu Arg Arg Met Ile Asp Gly Lys Ile Gln Phe Lys Ile Ser 85 90 95Met Pro Thr Ile Ala His Gly Asp Gly Arg Arg Pro Asn Lys Gln Arg 100 105 110Gln Tyr Ile Val Met Lys Ala Cys Asn Lys His His Ile Gly Ala Glu 115 120 125Ile Glu Leu Ala Ala Ala Asp Ile Glu Leu Leu Phe Ala Glu Lys Glu 130 135 140Thr Pro Leu Asp Phe Thr Glu Tyr Ala Gly Ala Ile Lys Thr Ile Thr145 150 155 160Ser Ala Leu Gln Phe Gly Met Asp Ala Leu Glu Arg Gly Leu Val Asp 165 170 175Thr Val Leu Ala Val Lys Leu Arg His Ala Pro Pro Val Phe Ile Leu 180 185 190Lys Thr Leu Gly Asp Pro Val Tyr Ser Glu Arg Gly Leu Lys Lys Ala 195 200 205Val Lys Ser Asp Met Val Ser Met Phe Lys Ala His Leu Ile Glu His 210 215 220Ser Phe Phe Leu Asp Lys Ala Glu Leu Met Thr Arg Gly Lys Gln Tyr225 230 235 240Val Leu Thr Met Leu Ser Asp Met Leu Ala Ala Val Cys Glu Asp Thr 245 250 255Val Phe Lys Gly Val Ser Thr Tyr Thr Thr Ala Ser Gly Gln Gln Val 260 265 270Ala Gly Val Leu Glu Thr Thr Asp Ser Val Met Arg Arg Leu Met Asn 275 280 285Leu Leu Gly Gln Val Glu Ser Ala Met Ser Gly Pro Ala Ala Tyr Ala 290 295 300Ser Tyr Val Val Arg Gly Ala Asn Leu Val Thr Ala Val Ser Tyr Gly305 310 315 320Arg Ala Met Arg Asn Phe Glu Gln Phe Met Ala Arg Ile Val Asp His 325 330 335Pro Asn Ala Leu Pro Ser Val Glu Gly Asp Lys Ala Ala Leu Ala Asp 340 345 350Gly His Asp Glu Ile Gln Arg Thr Arg Ile Ala Ala Ser Leu Val Lys 355 360 365Ile Gly Asp Lys Phe Val Ala Ile Glu Ser Leu Gln Arg Met Tyr Asn 370 375 380Glu Thr Gln Phe Pro Cys Pro Leu Asn Arg Arg Ile Gln Tyr Thr Tyr385 390 395 400Phe Phe Pro Val Gly Leu His Leu Pro Val Pro Arg Tyr Ser Thr Ser 405 410 415Val Ser Val Arg Gly Val Glu Ser Pro Ala Ile Gln Ser Thr Glu Thr 420 425 430Trp Val Val Asn Lys Asn Asn Val Pro Leu Cys Phe Gly Tyr Gln Asn 435 440 445Ala Leu Lys Ser Ile Cys His Pro Arg Met His Asn Pro Thr Gln Ser 450 455 460Ala Gln Ala Leu Asn Gln Ala Phe Pro Asp Pro Asp Gly Gly His Gly465 470 475 480Tyr Gly Leu Arg Tyr Glu Gln Thr Pro Asn Met Asn Leu Phe Arg Thr 485 490 495Phe His Gln Tyr Tyr Met Gly Lys Asn Val Ala Phe Val Pro Asp Val 500 505 510Ala Gln Lys Ala Leu Val Thr Thr Glu Asp Leu Leu His Pro Thr Ser 515 520 525His Arg Leu Leu Arg Leu Glu Val His Pro Phe Phe Asp Phe Phe Val 530 535 540His Pro Cys Pro Gly Ala Arg Gly Ser Tyr Arg Ala Thr His Arg Thr545 550 555 560Met Val Gly Asn Ile Pro Gln Pro Leu Ala Pro Arg Glu Phe Gln Glu 565 570 575Ser Arg Gly Ala Gln Phe Asp Ala Val Thr Asn Met Thr His Val Ile 580 585 590Asp Gln Leu Thr Ile Asp Val Ile Gln Glu Thr Ala Phe Asp Pro Ala 595 600 605Tyr Pro Leu Phe Cys Tyr Val Ile Glu Ala Met Ile His Gly Gln Glu 610 615 620Glu Lys Phe Val Met Asn Met Pro Leu Ile Ala Leu Val Ile Gln Thr625 630 635 640Tyr Trp Val Asn Ser Gly Lys Leu Ala Phe Val Asn Ser Tyr His Met 645 650 655Val Arg Phe Ile Cys Thr His Ile Gly Asn Gly Ser Ile Pro Lys Glu 660 665 670Ala His Gly His Tyr Arg Lys Ile Leu Gly Glu Leu Ile Ala Leu Glu 675 680 685Gln Ala Leu Leu Lys Leu Ala Gly His Glu Thr Val Gly Arg Thr Pro 690 695 700Ile Thr His Leu Val Ser Ala Leu Leu Asp Pro His Leu Leu Pro Pro705 710 715 720Phe Ala Tyr His Asp Val Phe Thr Asp Leu Met Gln Lys Ser Ser Arg 725 730 735Gln Pro Ile Ile Lys Ile Gly Asp Gln Asn Tyr Asp Asn Pro Gln Asn 740 745 750Arg Ala Thr Phe Ile Asn Leu Arg Gly Arg Met Glu Asp Leu Val Asn 755 760 765Asn Leu Val Asn Ile Tyr Gln Thr Arg Val Asn Glu Asp His Asp Glu 770 775 780Arg His Val Leu Asp Val Ala Pro Leu Asp Glu Asn Asp Tyr Asn Pro785 790 795 800Val Leu Glu Lys Leu Phe Tyr Tyr Val Leu Met Pro Val Cys Ser Asn 805 810 815Gly His Met Cys Gly Met Gly Val Asp Tyr Gln Asn Val Ala Leu Thr 820 825 830Leu
Thr Tyr Asn Gly Pro Val Phe Ala Asp Val Val Asn Ala Gln Asp 835 840 845Asp Ile Leu Leu His Leu Glu Asn Gly Thr Leu Lys Asp Ile Leu Gln 850 855 860Ala Gly Asp Ile Arg Pro Thr Val Asp Met Ile Arg Val Leu Cys Thr865 870 875 880Ser Phe Leu Thr Cys Pro Phe Val Thr Gln Ala Ala Arg Val Ile Thr 885 890 895Lys Arg Asp Pro Ala Gln Ser Phe Ala Thr His Glu Tyr Gly Lys Asp 900 905 910Val Ala Gln Thr Val Leu Val Asn Gly Phe Gly Ala Phe Ala Val Ala 915 920 925Asp Arg Ser Arg Glu Ala Ala Glu Thr Met Phe Tyr Pro Val Pro Phe 930 935 940Asn Lys Leu Tyr Ala Asp Pro Leu Val Ala Ala Thr Leu His Pro Leu945 950 955 960Leu Pro Asn Tyr Val Thr Arg Leu Pro Asn Gln Arg Asn Ala Val Val 965 970 975Phe Asn Val Pro Ser Asn Leu Met Ala Glu Tyr Glu Glu Trp His Lys 980 985 990Ser Pro Val Ala Ala Tyr Ala Ala Ser Cys Gln Ala Thr Pro Gly Ala 995 1000 1005Ile Ser Ala Met Val Ser Met His Gln Lys Leu Ser Ala Pro Ser 1010 1015 1020Phe Ile Cys Gln Ala Lys His Arg Met His Pro Gly Phe Ala Met 1025 1030 1035Thr Val Val Arg Thr Asp Glu Val Leu Ala Glu His Ile Leu Tyr 1040 1045 1050Cys Ser Arg Ala Ser Thr Ser Met Phe Val Gly Leu Pro Ser Val 1055 1060 1065Val Arg Arg Glu Val Arg Ser Asp Ala Val Thr Phe Glu Ile Thr 1070 1075 1080His Glu Ile Ala Ser Leu His Thr Ala Leu Gly Tyr Ser Ser Val 1085 1090 1095Ile Ala Pro Ala His Val Ala Ala Ile Thr Thr Asp Met Gly Val 1100 1105 1110His Cys Gln Asp Leu Phe Met Ile Phe Pro Gly Asp Ala Tyr Gln 1115 1120 1125Asp Arg Gln Leu His Asp Tyr Ile Lys Met Lys Ala Gly Val Gln 1130 1135 1140Thr Gly Ser Pro Gly Asn Arg Met Asp His Val Gly Tyr Thr Ala 1145 1150 1155Gly Val Pro Arg Cys Glu Asn Leu Pro Gly Leu Ser His Gly Gln 1160 1165 1170Leu Ala Thr Cys Glu Ile Ile Pro Thr Pro Val Thr Ser Asp Val 1175 1180 1185Ala Tyr Phe Gln Thr Pro Ser Asn Pro Arg Gly Arg Ala Ala Ser 1190 1195 1200Val Val Ser Cys Asp Ala Tyr Ser Asn Glu Ser Ala Glu Arg Leu 1205 1210 1215Phe Tyr Asp His Ser Ile Pro Asp Pro Ala Tyr Glu Cys Arg Ser 1220 1225 1230Thr Asn Asn Pro Trp Ala Ser Gln Arg Gly Ser Leu Gly Asp Val 1235 1240 1245Leu Tyr Asn Ile Thr Phe Arg Gln Thr Ala Leu Pro Gly Met Tyr 1250 1255 1260Ser Pro Cys Arg Gln Phe Phe His Lys Glu Asp Ile Met Arg Tyr 1265 1270 1275Asn Arg Gly Leu Tyr Thr Leu Val Asn Glu Tyr Ser Ala Arg Leu 1280 1285 1290Ala Gly Ala Pro Ala Thr Ser Thr Thr Asp Leu Gln Tyr Val Val 1295 1300 1305Val Asn Gly Thr Asp Val Phe Leu Asp Gln Pro Cys His Met Leu 1310 1315 1320Gln Glu Ala Tyr Pro Thr Leu Ala Ala Ser His Arg Val Met Leu 1325 1330 1335Ala Glu Tyr Met Ser Asn Lys Gln Thr His Ala Pro Val His Met 1340 1345 1350Gly Gln Tyr Leu Ile Glu Glu Val Ala Pro Met Lys Arg Leu Leu 1355 1360 1365Lys Leu Gly Asn Lys Val Val Tyr 1370 137541143DNAKaposi's sarcoma-associated herpesvirus 4agcattcggg gacagacctt taacctgctc tacgtagacg aggcgaattt tattaaaaag 60gatgcactgc cggctattct gggtttcatg cttcagaaag acgccaagct tatatttata 120tcatccgtga actcgtcaga ccgctccacg agtttcctgc ttaacctcag gaacgcccag 180gaaaagatgc tgaatgtggt cagttacgtg tgtgcggacc accgagaaga tttccacctg 240caagacgcac tagtgtcctg tccttgttac agactgcaca ttccgacgta catcaccatc 300gacgaatcca tcaaaaccac caccaacctc tttatggagg gggcattcga caccgaacta 360atgggcgagg gagcagcgtc gtcaaatgct acgctttacc gcgtggtggg tgacgcagcg 420ctgacacagt ttgacatgtg tcgggtagac accaccgccc aggaggttca gaagtgcctt 480ggaaaacagc tgtttgttta catcgacccc gcgtatacga acaacacgga ggcgtccggt 540actggcgtgg gcgccgttgt cacgagtact cagactccca ccagaagcct catattgggc 600atggagcatt tcttcctgcg cgacctcact ggcgcagctg cttacgagat agcgtcctgc 660gcatgcacga tgattaaggc gatcgctgtg ctccacacca caattgagcg cgtgaacgcg 720gcggtcgaag gcaacagcag ccaagattct ggggtggcca ttgcaaccgt ccttaacgaa 780atatgcccgc tccccataca ttttctacac tatactgaca agagcagcgc cctgcagtgg 840ccaatttaca tgttgggagg cgagaaatcc tccgcgtttg agacattcat ctacgctctg 900aactccggca ccctgagcgc cagccagacg gtggtgtcca acaccatcaa aatatcattt 960gacccggtga cctacctggt agaacaggtc cgcgcgatca agtgcgtccc gcttagggat 1020ggagggcagt catacagcgc caagcaaaag cacatgtcgg acgacttact tgtggcagtt 1080gtcatggccc attttatggc taccgatgat agacacatgt acaagcccat atccccacaa 1140taa 11435380PRTKaposi's sarcoma-associated herpesvirus 5Ser Ile Arg Gly Gln Thr Phe Asn Leu Leu Tyr Val Asp Glu Ala Asn1 5 10 15Phe Ile Lys Lys Asp Ala Leu Pro Ala Ile Leu Gly Phe Met Leu Gln 20 25 30Lys Asp Ala Lys Leu Ile Phe Ile Ser Ser Val Asn Ser Ser Asp Arg 35 40 45Ser Thr Ser Phe Leu Leu Asn Leu Arg Asn Ala Gln Glu Lys Met Leu 50 55 60Asn Val Val Ser Tyr Val Cys Ala Asp His Arg Glu Asp Phe His Leu65 70 75 80Gln Asp Ala Leu Val Ser Cys Pro Cys Tyr Arg Leu His Ile Pro Thr 85 90 95Tyr Ile Thr Ile Asp Glu Ser Ile Lys Thr Thr Thr Asn Leu Phe Met 100 105 110Glu Gly Ala Phe Asp Thr Glu Leu Met Gly Glu Gly Ala Ala Ser Ser 115 120 125Asn Ala Thr Leu Tyr Arg Val Val Gly Asp Ala Ala Leu Thr Gln Phe 130 135 140Asp Met Cys Arg Val Asp Thr Thr Ala Gln Glu Val Gln Lys Cys Leu145 150 155 160Gly Lys Gln Leu Phe Val Tyr Ile Asp Pro Ala Tyr Thr Asn Asn Thr 165 170 175Glu Ala Ser Gly Thr Gly Val Gly Ala Val Val Thr Ser Thr Gln Thr 180 185 190Pro Thr Arg Ser Leu Ile Leu Gly Met Glu His Phe Phe Leu Arg Asp 195 200 205Leu Thr Gly Ala Ala Ala Tyr Glu Ile Ala Ser Cys Ala Cys Thr Met 210 215 220Ile Lys Ala Ile Ala Val Leu His Thr Thr Ile Glu Arg Val Asn Ala225 230 235 240Ala Val Glu Gly Asn Ser Ser Gln Asp Ser Gly Val Ala Ile Ala Thr 245 250 255Val Leu Asn Glu Ile Cys Pro Leu Pro Ile His Phe Leu His Tyr Thr 260 265 270Asp Lys Ser Ser Ala Leu Gln Trp Pro Ile Tyr Met Leu Gly Gly Glu 275 280 285Lys Ser Ser Ala Phe Glu Thr Phe Ile Tyr Ala Leu Asn Ser Gly Thr 290 295 300Leu Ser Ala Ser Gln Thr Val Val Ser Asn Thr Ile Lys Ile Ser Phe305 310 315 320Asp Pro Val Thr Tyr Leu Val Glu Gln Val Arg Ala Ile Lys Cys Val 325 330 335Pro Leu Arg Asp Gly Gly Gln Ser Tyr Ser Ala Lys Gln Lys His Met 340 345 350Ser Asp Asp Leu Leu Val Ala Val Val Met Ala His Phe Met Ala Thr 355 360 365Asp Asp Arg His Met Tyr Lys Pro Ile Ser Pro Gln 370 375 3806234DNAKaposi's sarcoma-associated herpesvirus 6atgggtgagc cagtggatcc tggacatgtg gtgaatgaga aagattttga ggagtgtgaa 60caatttttca gtcaacccct tagggagcaa gtggtcgcgg gggtcagggc actcgacggc 120ctcggtctcg ctgactctct atgtcacaaa acagaaagac tctgcctgct gatggacctg 180gtgggcacgg agtgctttgc gagggtgtgc cgcctagaca ccggtgcgaa atga 234777PRTKaposi's sarcoma-associated herpesvirus 7Met Gly Glu Pro Val Asp Pro Gly His Val Val Asn Glu Lys Asp Phe1 5 10 15Glu Glu Cys Glu Gln Phe Phe Ser Gln Pro Leu Arg Glu Gln Val Val 20 25 30Ala Gly Val Arg Ala Leu Asp Gly Leu Gly Leu Ala Asp Ser Leu Cys 35 40 45His Lys Thr Glu Arg Leu Cys Leu Leu Met Asp Leu Val Gly Thr Glu 50 55 60Cys Phe Ala Arg Val Cys Arg Leu Asp Thr Gly Ala Lys65 70 758585DNAKaposi's sarcoma-associated herpesvirus 8atgaagagtg tggcgagtcc cttatgtcag ttccacggcg tgttttgcct gtaccagtgt 60cgccagtgcc tggcatacca cgtgtgtgat gggggcgccg aatgcgttct cctgcatacg 120ccggagagcg tcatctgcga actaacgggt aactgcatgc tcggcaacat tcaagagggc 180cagtttttag ggccggtacc gtatcggact ttggataacc aggttgacag ggacgcatat 240cacgggatgc tagcgtgtct gaaacgggac attgtgcggt atttgcagac atggccggac 300accaccgtaa tcgtgcagga aatagccctg ggggacggcg tcaccgacac catctcggcc 360attatagatg aaacattcgg tgagtgtctt cccgtactgg gggaggccca aggcgggtac 420gccctggtct gtagcatgta tctgcacgtt atcgtctcca tctattcgac aaaaacggtg 480tacaacagta tgctatttaa atgcacaaag aataaaaagt acgactgcat tgccaagcgg 540gtgcggacaa aatggatgcg catgctatca acgaaagata cgtag 5859194PRTKaposi's sarcoma-assciated herpesvirus 9Met Lys Ser Val Ala Ser Pro Leu Cys Gln Phe His Gly Val Phe Cys1 5 10 15Leu Tyr Gln Cys Arg Gln Cys Leu Ala Tyr His Val Cys Asp Gly Gly 20 25 30Ala Glu Cys Val Leu Leu His Thr Pro Glu Ser Val Ile Cys Glu Leu 35 40 45Thr Gly Asn Cys Met Leu Gly Asn Ile Gln Glu Gly Gln Phe Leu Gly 50 55 60Pro Val Pro Tyr Arg Thr Leu Asp Asn Gln Val Asp Arg Asp Ala Tyr65 70 75 80His Gly Met Leu Ala Cys Leu Lys Arg Asp Ile Val Arg Tyr Leu Gln 85 90 95Thr Trp Pro Asp Thr Thr Val Ile Val Gln Glu Ile Ala Leu Gly Asp 100 105 110Gly Val Thr Asp Thr Ile Ser Ala Ile Ile Asp Glu Thr Phe Gly Glu 115 120 125Cys Leu Pro Val Leu Gly Glu Ala Gln Gly Gly Tyr Ala Leu Val Cys 130 135 140Ser Met Tyr Leu His Val Ile Val Ser Ile Tyr Ser Thr Lys Thr Val145 150 155 160Tyr Asn Ser Met Leu Phe Lys Cys Thr Lys Asn Lys Lys Tyr Asp Cys 165 170 175Ile Ala Lys Arg Val Arg Thr Lys Trp Met Arg Met Leu Ser Thr Lys 180 185 190Asp Thr 10939DNAKaposi's sarcoma-associated herpesvirus 10atggctagcc ggaggcgcaa acttcggaat ttcctaaaca aggaatgcat atggactgtt 60aacccaatgt caggggacca tatcaaggtc tttaacgcct gcacctctat ctcgccggtg 120tatgaccctg agctggtaac cagctacgca ctgagcgtgc ctgcttacaa tgtgtctgtg 180gctatcttgc tgcataaagt catgggaccg tgtgtggctg tgggaattaa cggagaaatg 240atcatgtacg tcgtaagcca gtgtgtttct gtgcggcccg tcccggggcg cgatggtatg 300gcgctcatct actttggaca gtttctggag gaagcatccg gactgagatt tccctacatt 360gctccgccgc cgtcgcgcga acacgtacct gacctgacca gacaagaatt agttcatacc 420tcccaggtgg tgcgccgcgg cgacctgacc aattgcacta tgggtctcga attcaggaat 480gtgaaccctt ttgtttggct cgggggcgga tcggtgtggc tgctgttctt gggcgtggac 540tacatggcgt tctgtccggg tgtcgacgga atgccgtcgt tggcaagagt ggccgccctg 600cttaccaggt gcgaccaccc agactgtgtc cactgccatg gactccgtgg acacgttaat 660gtatttcgtg ggtactgttc tgcgcagtcg ccgggtctat ctaacatctg tccctgtatc 720aaatcatgtg ggaccgggaa tggagtgact agggtcactg gaaacagaaa ttttctgggt 780cttctgttcg atcccattgt ccagagcagg gtaacagctc tgaagataac tagccaccca 840acccccacgc acgtcgagaa tgtgctaaca ggagtgctcg acgacggcac cttggtgccg 900tccgtccaag gcaccctggg tcctcttacg aatgtctga 93911312PRTKaposi's sarcoma-associated herpesvirus 11Met Ala Ser Arg Arg Arg Lys Leu Arg Asn Phe Leu Asn Lys Glu Cys1 5 10 15Ile Trp Thr Val Asn Pro Met Ser Gly Asp His Ile Lys Val Phe Asn 20 25 30Ala Cys Thr Ser Ile Ser Pro Val Tyr Asp Pro Glu Leu Val Thr Ser 35 40 45Tyr Ala Leu Ser Val Pro Ala Tyr Asn Val Ser Val Ala Ile Leu Leu 50 55 60His Lys Val Met Gly Pro Cys Val Ala Val Gly Ile Asn Gly Glu Met65 70 75 80Ile Met Tyr Val Val Ser Gln Cys Val Ser Val Arg Pro Val Pro Gly 85 90 95Arg Asp Gly Met Ala Leu Ile Tyr Phe Gly Gln Phe Leu Glu Glu Ala 100 105 110Ser Gly Leu Arg Phe Pro Tyr Ile Ala Pro Pro Pro Ser Arg Glu His 115 120 125Val Pro Asp Leu Thr Arg Gln Glu Leu Val His Thr Ser Gln Val Val 130 135 140Arg Arg Gly Asp Leu Thr Asn Cys Thr Met Gly Leu Glu Phe Arg Asn145 150 155 160Val Asn Pro Phe Val Trp Leu Gly Gly Gly Ser Val Trp Leu Leu Phe 165 170 175Leu Gly Val Asp Tyr Met Ala Phe Cys Pro Gly Val Asp Gly Met Pro 180 185 190Ser Leu Ala Arg Val Ala Ala Leu Leu Thr Arg Cys Asp His Pro Asp 195 200 205Cys Val His Cys His Gly Leu Arg Gly His Val Asn Val Phe Arg Gly 210 215 220Tyr Cys Ser Ala Gln Ser Pro Gly Leu Ser Asn Ile Cys Pro Cys Ile225 230 235 240Lys Ser Cys Gly Thr Gly Asn Gly Val Thr Arg Val Thr Gly Asn Arg 245 250 255Asn Phe Leu Gly Leu Leu Phe Asp Pro Ile Val Gln Ser Arg Val Thr 260 265 270Ala Leu Lys Ile Thr Ser His Pro Thr Pro Thr His Val Glu Asn Val 275 280 285Leu Thr Gly Val Leu Asp Asp Gly Thr Leu Val Pro Ser Val Gln Gly 290 295 300Thr Leu Gly Pro Leu Thr Asn Val305 3101286DNAKaposi's sarcoma-associated herpesvirus 12atggactcaa ccaactctaa aagagagttt attaagtcgg ctctggaggc caacatcaac 60aggagggcag ctgtatcgct atttga 861328PRTKaposi's sarcoma-associated herpesvirus 13Met Asp Ser Thr Asn Ser Lys Arg Glu Phe Ile Lys Ser Ala Leu Glu1 5 10 15Ala Asn Ile Asn Arg Arg Ala Ala Val Ser Leu Phe 20 25141743DNAKaposi's sarcoma-associated herpesvirus 14atggcagaag gcggttttgg agcggactcg gtggggcgcg gcggagaaaa ggcctctgtg 60actaggggag gcaggtggga cttggggagc tcggacgacg aatcaagcac ctccacaacc 120agcacggata tggacgacct ccctgaggag aggaaaccac taacgggaaa gtctgtaaaa 180acctcgtaca tatacgacgt gcccaccgtc ccgaccagca agccgtggca tttaatgcac 240gacaactccc tctacgcaac gcctaggttt ccgcccagac ctctcatacg gcacccttcc 300gaaaaaggca gcatttttgc cagtcggttg tcagcgactg acgacgactc gggagactac 360gcgccaatgg atcgcttcgc cttccagagc cccagggtgt gtggtcgccc tccccttccg 420cctccaaatc acccacctcc ggcaactagg ccggcagacg cgtcaatggg ggacgtgggc 480tgggcggatc tgcagggact caagaggacc ccaaagggat ttttaaaaac atctaccaag 540gggggcagtc tcaaagcccg tggacgcgat gtaggtgacc gtctcaggga cggcggcttt 600gcctttagtc ctaggggcgt gaaatctgcc atagggcaaa acattaaatc atggttgggg 660atcggagaat catcggcgac tgctgtcccc gtcaccacgc agcttatggt accggtgcac 720ctcattagaa cgcctgtgac cgtggactac aggaatgttt atttgcttta cttagagggg 780gtaatgggtg tgggcaaatc aacgctggtc aacgccgtgt gcgggatctt gccccaggag 840agagtgacaa gttttcccga gcccatggtg tactggacga gggcatttac agattgttac 900aaggaaattt cccacctgat gaagtctggt aaggcgggag acccgctgac gtctgccaaa 960atatactcat gccaaaacaa gttttcgctc cccttccgga cgaacgccac cgctatcctg 1020cgaatgatgc agccctggaa cgttgggggt gggtctggga ggggcactca ctggtgcgtc 1080tttgataggc atctcctctc cccagcagtg gtgttccctc tcatgcacct gaagcacggc 1140cgcctatctt ttgatcactt ctttcaatta ctttccatct ttagagccac agaaggcgac 1200gtggtcgcca ttctcaccct ctccagcgcc gagtcgttgc ggcgggtcag ggcgagggga 1260agaaagaacg acgggacggt ggagcaaaac tacatcagag aattggcgtg ggcttatcac 1320gccgtgtact gttcatggat catgttgcag tacatcactg tggagcagat ggtacaacta 1380tgcgtacaaa ccacaaatat tccggaaatc tgcttccgca gcgtgcgcct ggcacacaag 1440gaggaaactt tgaaaaacct tcacgagcag agcatgctac ctatgatcac cggtgtactg 1500gatcccgtga gacatcatcc cgtcgtgatc gagctttgct tttgtttctt cacagagctg 1560agaaaattac aatttatcgt agccgacgcg gataagttcc acgacgacgt atgcggcctg 1620tggaccgaaa tctacaggca gatcctgtcc aatccggcta ttaaacccag ggccatcaac 1680tggccagcat tagagagcca gtctaaagca gttaatcacc tagaggagac atgcagggtc 1740tag 174315580PRTKaposi's sarcoma-associated herpesvirus 15Met Ala Glu Gly Gly Phe Gly Ala Asp Ser Val Gly Arg Gly Gly Glu1 5 10 15Lys Ala Ser Val Thr Arg Gly Gly Arg Trp Asp Leu Gly Ser Ser Asp 20 25 30Asp Glu Ser Ser Thr Ser Thr Thr Ser Thr Asp Met Asp Asp Leu Pro
35 40 45Glu Glu Arg Lys Pro Leu Thr Gly Lys Ser Val Lys Thr Ser Tyr Ile 50 55 60Tyr Asp Val Pro Thr Val Pro Thr Ser Lys Pro Trp His Leu Met His65 70 75 80Asp Asn Ser Leu Tyr Ala Thr Pro Arg Phe Pro Pro Arg Pro Leu Ile 85 90 95Arg His Pro Ser Glu Lys Gly Ser Ile Phe Ala Ser Arg Leu Ser Ala 100 105 110Thr Asp Asp Asp Ser Gly Asp Tyr Ala Pro Met Asp Arg Phe Ala Phe 115 120 125Gln Ser Pro Arg Val Cys Gly Arg Pro Pro Leu Pro Pro Pro Asn His 130 135 140Pro Pro Pro Ala Thr Arg Pro Ala Asp Ala Ser Met Gly Asp Val Gly145 150 155 160Trp Ala Asp Leu Gln Gly Leu Lys Arg Thr Pro Lys Gly Phe Leu Lys 165 170 175Thr Ser Thr Lys Gly Gly Ser Leu Lys Ala Arg Gly Arg Asp Val Gly 180 185 190Asp Arg Leu Arg Asp Gly Gly Phe Ala Phe Ser Pro Arg Gly Val Lys 195 200 205Ser Ala Ile Gly Gln Asn Ile Lys Ser Trp Leu Gly Ile Gly Glu Ser 210 215 220Ser Ala Thr Ala Val Pro Val Thr Thr Gln Leu Met Val Pro Val His225 230 235 240Leu Ile Arg Thr Pro Val Thr Val Asp Tyr Arg Asn Val Tyr Leu Leu 245 250 255Tyr Leu Glu Gly Val Met Gly Val Gly Lys Ser Thr Leu Val Asn Ala 260 265 270Val Cys Gly Ile Leu Pro Gln Glu Arg Val Thr Ser Phe Pro Glu Pro 275 280 285Met Val Tyr Trp Thr Arg Ala Phe Thr Asp Cys Tyr Lys Glu Ile Ser 290 295 300His Leu Met Lys Ser Gly Lys Ala Gly Asp Pro Leu Thr Ser Ala Lys305 310 315 320Ile Tyr Ser Cys Gln Asn Lys Phe Ser Leu Pro Phe Arg Thr Asn Ala 325 330 335Thr Ala Ile Leu Arg Met Met Gln Pro Trp Asn Val Gly Gly Gly Ser 340 345 350Gly Arg Gly Thr His Trp Cys Val Phe Asp Arg His Leu Leu Ser Pro 355 360 365Ala Val Val Phe Pro Leu Met His Leu Lys His Gly Arg Leu Ser Phe 370 375 380Asp His Phe Phe Gln Leu Leu Ser Ile Phe Arg Ala Thr Glu Gly Asp385 390 395 400Val Val Ala Ile Leu Thr Leu Ser Ser Ala Glu Ser Leu Arg Arg Val 405 410 415Arg Ala Arg Gly Arg Lys Asn Asp Gly Thr Val Glu Gln Asn Tyr Ile 420 425 430Arg Glu Leu Ala Trp Ala Tyr His Ala Val Tyr Cys Ser Trp Ile Met 435 440 445Leu Gln Tyr Ile Thr Val Glu Gln Met Val Gln Leu Cys Val Gln Thr 450 455 460Thr Asn Ile Pro Glu Ile Cys Phe Arg Ser Val Arg Leu Ala His Lys465 470 475 480Glu Glu Thr Leu Lys Asn Leu His Glu Gln Ser Met Leu Pro Met Ile 485 490 495Thr Gly Val Leu Asp Pro Val Arg His His Pro Val Val Ile Glu Leu 500 505 510Cys Phe Cys Phe Phe Thr Glu Leu Arg Lys Leu Gln Phe Ile Val Ala 515 520 525Asp Ala Asp Lys Phe His Asp Asp Val Cys Gly Leu Trp Thr Glu Ile 530 535 540Tyr Arg Gln Ile Leu Ser Asn Pro Ala Ile Lys Pro Arg Ala Ile Asn545 550 555 560Trp Pro Ala Leu Glu Ser Gln Ser Lys Ala Val Asn His Leu Glu Glu 565 570 575Thr Cys Arg Val 580162193DNAKaposi's sarcoma-associated herpesvirus 16atgcagggtc tagccttctt ggcggccctt gcatgctggc gatgcatatc gttgacatgt 60ggagccactg gcgcgttgcc gacaacggcg acgacaataa cccgctccgc cacgcagctc 120atcaatggga gaaccaacct ctccatagaa ctggaattca acggcactag tttttttcta 180aattggcaaa atctgttgaa tgtgatcacg gagccggccc tgacagagtt gtggacctcc 240gccgaagtcg ccgaggacct cagggtaact ctgaaaaaga ggcaaagtct ttttttcccc 300aacaagacag ttgtgatctc tggagacggc catcgctata cgtgcgaggt gccgacgtcg 360tcgcaaactt ataacatcac caagggcttt aactatagcg ctctgcccgg gcaccttggc 420ggatttggga tcaacgcgcg tctggtactg ggtgatatct tcgcatcaaa atggtcgcta 480ttcgcgaggg acaccccaga gtatcgggtg ttttacccaa tgaatgtcat ggccgtcaag 540ttttccatat ccattggcaa caacgagtcc ggcgtagcgc tctatggagt ggtgtcggaa 600gatttcgtgg tcgtcacgct ccacaacagg tccaaagagg ctaacgagac ggcgtcccat 660cttctgttcg gtctcccgga ttcactgcca tctctgaagg gccatgccac ctatgatgaa 720ctcacgttcg cccgaaacgc aaaatatgcg ctagtggcga tcctgcctaa agattcttac 780cagacactcc ttacagagaa ttacactcgc atatttctga acatgacgga gtcgacgccc 840ctcgagttca cgcggacgat ccagaccagg atcgtatcaa tcgaggccag gcgcgcctgc 900gcagctcaag aggcggcgcc ggacatattc ttggtgttgt ttcagatgtt ggtggcacac 960tttcttgttg cgcggggcat tgccgagcac cgatttgtgg aggtggactg cgtgtgtcgg 1020cagtatgcgg aactgtattt tctccgccgc atctcgcgtc tgtgcatgcc cacgttcacc 1080actgtcgggt ataaccacac cacccttggc gctgtggccg ccacacaaat agctcgcgtg 1140tccgccacga agttggccag tttgccccgc tcttcccagg aaacagtgct ggccatggtc 1200cagcttggcg cccgtgatgg cgccgtccct tcctccattc tggagggcat tgctatggtc 1260gtcgaacata tgtataccgc ctacacttat gtgtacacac tcggcgatac tgaaagaaaa 1320ttaatgttgg acatacacac ggtcctcacc gacagctgcc cgcccaaaga ctccggagta 1380tcagaaaagc tactgagaac atatttgatg ttcacatcaa tgtgtaccaa catagagctg 1440ggcgaaatga tcgcccgctt ttccaaaccg gacagcctta acatctatag ggcattctcc 1500ccctgctttc taggactaag gtacgatttg catccagcca agttgcgcgc cgaggcgccg 1560cagtcgtccg ctctgacgcg gactgccgtt gccagaggaa catcgggatt cgcagaattg 1620ctccacgcgc tgcacctcga tagcttaaat ttaattccgg cgattaactg ttcaaagatt 1680acagccgaca agataatagc tacggtaccc ttgcctcacg tcacgtatat catcagttcc 1740gaagcactct cgaacgctgt tgtctacgag gtgtcggaga tcttcctcaa gagtgccatg 1800tttatatctg ctatcaaacc cgattgctcc ggctttaact tttctcagat tgataggcac 1860attcccatag tctacaacat cagcacacca agaagaggtt gccccctttg tgactctgta 1920atcatgagct acgatgagag cgatggcctg cagtctctca tgtatgtcac taatgaaagg 1980gtgcagacca acctcttttt agataagtca cctttctttg ataataacaa cctacacatt 2040cattatttgt ggctgaggga caacgggacc gtagtggaga taaggggcat gtatagaaga 2100cgcgcagcca gtgctttgtt tctaattctc tcttttattg ggttctcggg ggttatctac 2160tttctttaca gactgttttc catcctttat tag 219317730PRTKaposi's sarcoma-associated herpesvirus 17Met Gln Gly Leu Ala Phe Leu Ala Ala Leu Ala Cys Trp Arg Cys Ile1 5 10 15Ser Leu Thr Cys Gly Ala Thr Gly Ala Leu Pro Thr Thr Ala Thr Thr 20 25 30Ile Thr Arg Ser Ala Thr Gln Leu Ile Asn Gly Arg Thr Asn Leu Ser 35 40 45Ile Glu Leu Glu Phe Asn Gly Thr Ser Phe Phe Leu Asn Trp Gln Asn 50 55 60Leu Leu Asn Val Ile Thr Glu Pro Ala Leu Thr Glu Leu Trp Thr Ser65 70 75 80Ala Glu Val Ala Glu Asp Leu Arg Val Thr Leu Lys Lys Arg Gln Ser 85 90 95Leu Phe Phe Pro Asn Lys Thr Val Val Ile Ser Gly Asp Gly His Arg 100 105 110Tyr Thr Cys Glu Val Pro Thr Ser Ser Gln Thr Tyr Asn Ile Thr Lys 115 120 125Gly Phe Asn Tyr Ser Ala Leu Pro Gly His Leu Gly Gly Phe Gly Ile 130 135 140Asn Ala Arg Leu Val Leu Gly Asp Ile Phe Ala Ser Lys Trp Ser Leu145 150 155 160Phe Ala Arg Asp Thr Pro Glu Tyr Arg Val Phe Tyr Pro Met Asn Val 165 170 175Met Ala Val Lys Phe Ser Ile Ser Ile Gly Asn Asn Glu Ser Gly Val 180 185 190Ala Leu Tyr Gly Val Val Ser Glu Asp Phe Val Val Val Thr Leu His 195 200 205Asn Arg Ser Lys Glu Ala Asn Glu Thr Ala Ser His Leu Leu Phe Gly 210 215 220Leu Pro Asp Ser Leu Pro Ser Leu Lys Gly His Ala Thr Tyr Asp Glu225 230 235 240Leu Thr Phe Ala Arg Asn Ala Lys Tyr Ala Leu Val Ala Ile Leu Pro 245 250 255Lys Asp Ser Tyr Gln Thr Leu Leu Thr Glu Asn Tyr Thr Arg Ile Phe 260 265 270Leu Asn Met Thr Glu Ser Thr Pro Leu Glu Phe Thr Arg Thr Ile Gln 275 280 285Thr Arg Ile Val Ser Ile Glu Ala Arg Arg Ala Cys Ala Ala Gln Glu 290 295 300Ala Ala Pro Asp Ile Phe Leu Val Leu Phe Gln Met Leu Val Ala His305 310 315 320Phe Leu Val Ala Arg Gly Ile Ala Glu His Arg Phe Val Glu Val Asp 325 330 335Cys Val Cys Arg Gln Tyr Ala Glu Leu Tyr Phe Leu Arg Arg Ile Ser 340 345 350Arg Leu Cys Met Pro Thr Phe Thr Thr Val Gly Tyr Asn His Thr Thr 355 360 365Leu Gly Ala Val Ala Ala Thr Gln Ile Ala Arg Val Ser Ala Thr Lys 370 375 380Leu Ala Ser Leu Pro Arg Ser Ser Gln Glu Thr Val Leu Ala Met Val385 390 395 400Gln Leu Gly Ala Arg Asp Gly Ala Val Pro Ser Ser Ile Leu Glu Gly 405 410 415Ile Ala Met Val Val Glu His Met Tyr Thr Ala Tyr Thr Tyr Val Tyr 420 425 430Thr Leu Gly Asp Thr Glu Arg Lys Leu Met Leu Asp Ile His Thr Val 435 440 445Leu Thr Asp Ser Cys Pro Pro Lys Asp Ser Gly Val Ser Glu Lys Leu 450 455 460Leu Arg Thr Tyr Leu Met Phe Thr Ser Met Cys Thr Asn Ile Glu Leu465 470 475 480Gly Glu Met Ile Ala Arg Phe Ser Lys Pro Asp Ser Leu Asn Ile Tyr 485 490 495Arg Ala Phe Ser Pro Cys Phe Leu Gly Leu Arg Tyr Asp Leu His Pro 500 505 510Ala Lys Leu Arg Ala Glu Ala Pro Gln Ser Ser Ala Leu Thr Arg Thr 515 520 525Ala Val Ala Arg Gly Thr Ser Gly Phe Ala Glu Leu Leu His Ala Leu 530 535 540His Leu Asp Ser Leu Asn Leu Ile Pro Ala Ile Asn Cys Ser Lys Ile545 550 555 560Thr Ala Asp Lys Ile Ile Ala Thr Val Pro Leu Pro His Val Thr Tyr 565 570 575Ile Ile Ser Ser Glu Ala Leu Ser Asn Ala Val Val Tyr Glu Val Ser 580 585 590Glu Ile Phe Leu Lys Ser Ala Met Phe Ile Ser Ala Ile Lys Pro Asp 595 600 605Cys Ser Gly Phe Asn Phe Ser Gln Ile Asp Arg His Ile Pro Ile Val 610 615 620Tyr Asn Ile Ser Thr Pro Arg Arg Gly Cys Pro Leu Cys Asp Ser Val625 630 635 640Ile Met Ser Tyr Asp Glu Ser Asp Gly Leu Gln Ser Leu Met Tyr Val 645 650 655Thr Asn Glu Arg Val Gln Thr Asn Leu Phe Leu Asp Lys Ser Pro Phe 660 665 670Phe Asp Asn Asn Asn Leu His Ile His Tyr Leu Trp Leu Arg Asp Asn 675 680 685Gly Thr Val Val Glu Ile Arg Gly Met Tyr Arg Arg Arg Ala Ala Ser 690 695 700Ala Leu Phe Leu Ile Leu Ser Phe Ile Gly Phe Ser Gly Val Ile Tyr705 710 715 720Phe Leu Tyr Arg Leu Phe Ser Ile Leu Tyr 725 730181215DNAKaposi's sarcoma-associated herpesvirus 18atgttacgag ttccggacgt gaaggctagt ctagtagagg gcgcggcgcg cctgtcgaca 60ggcgagcgcg tgtttcacgt cttgacctct ccggcggtgg cggccatggt gggagtctct 120aatcctgaag tcccgatgcc actgttgttc gaaaagtttg ggactccgga ctcgtctacc 180ctgccactct acgcggctag gcacccggaa ctatcgttgc tacggatcat gctctcaccg 240cacccctacg cgttaagaag ccacttgtgc gtaggcgaag agaccgcatc tcttggcgtt 300tacctgcact ccaagccagt cgtacgcggc cacgaattcg aggacacgca gatactaccg 360gagtgccggc tggccataac gagcgaccag tcttatacca actttaagat tatagatctg 420ccagcgggat gccgtcgcgt ccccatacac gccgcgaaca agcgtgtcgt catcgacgag 480gccgccaacc gcataaaggt gtttgaccca gagtcgcctt taccgcgtca ccccataaca 540ccccgtgccg gtcagaccag atctatactg aaacacaaca tcgcacaggt ttgcgaacgg 600gatatcgtgt cacttaacac agacaacgag gccgcgtcta tgttctacat gattggactc 660aggcggccga gactcggaga aagcccggtc tgtgacttca acaccgttac catcatggag 720cgtgctaaca actcgataac ttttctaccc aagctaaaac tgaaccggct acaacacctg 780ttcctgaagc acgtgttgct gcgcagcatg gggctggaaa acatcgtgtc gtgtttctca 840tcgctgtacg gcgcagaact tgcccctgcg aaaacacacg agcgggagtt cttcggcgct 900ctgctagaaa gactcaaacg tcgggtggag gacgcggtct tctgcctgaa taccatagag 960gatttcccgt ttagggaacc cattcgccaa cccccagatt gttccaaggt gcttatagaa 1020gccatggaaa agtactttat gatgtgtagc cccaaagacc gtcaaagcgc cgcatggcta 1080ggtgcagggg tggtcgaact gatatgtgac ggcaatccac tttctgaggt gctcggattt 1140cttgccaagt atatgcccat acaaaaagaa tgcacaggaa accttttaaa aatctacgct 1200ttattgaccg tctaa 121519404PRTKaposi's sarcoma-associated herpesvirus 19Met Leu Arg Val Pro Asp Val Lys Ala Ser Leu Val Glu Gly Ala Ala1 5 10 15Arg Leu Ser Thr Gly Glu Arg Val Phe His Val Leu Thr Ser Pro Ala 20 25 30Val Ala Ala Met Val Gly Val Ser Asn Pro Glu Val Pro Met Pro Leu 35 40 45Leu Phe Glu Lys Phe Gly Thr Pro Asp Ser Ser Thr Leu Pro Leu Tyr 50 55 60Ala Ala Arg His Pro Glu Leu Ser Leu Leu Arg Ile Met Leu Ser Pro65 70 75 80His Pro Tyr Ala Leu Arg Ser His Leu Cys Val Gly Glu Glu Thr Ala 85 90 95Ser Leu Gly Val Tyr Leu His Ser Lys Pro Val Val Arg Gly His Glu 100 105 110Phe Glu Asp Thr Gln Ile Leu Pro Glu Cys Arg Leu Ala Ile Thr Ser 115 120 125Asp Gln Ser Tyr Thr Asn Phe Lys Ile Ile Asp Leu Pro Ala Gly Cys 130 135 140Arg Arg Val Pro Ile His Ala Ala Asn Lys Arg Val Val Ile Asp Glu145 150 155 160Ala Ala Asn Arg Ile Lys Val Phe Asp Pro Glu Ser Pro Leu Pro Arg 165 170 175His Pro Ile Thr Pro Arg Ala Gly Gln Thr Arg Ser Ile Leu Lys His 180 185 190Asn Ile Ala Gln Val Cys Glu Arg Asp Ile Val Ser Leu Asn Thr Asp 195 200 205Asn Glu Ala Ala Ser Met Phe Tyr Met Ile Gly Leu Arg Arg Pro Arg 210 215 220Leu Gly Glu Ser Pro Val Cys Asp Phe Asn Thr Val Thr Ile Met Glu225 230 235 240Arg Ala Asn Asn Ser Ile Thr Phe Leu Pro Lys Leu Lys Leu Asn Arg 245 250 255Leu Gln His Leu Phe Leu Lys His Val Leu Leu Arg Ser Met Gly Leu 260 265 270Glu Asn Ile Val Ser Cys Phe Ser Ser Leu Tyr Gly Ala Glu Leu Ala 275 280 285Pro Ala Lys Thr His Glu Arg Glu Phe Phe Gly Ala Leu Leu Glu Arg 290 295 300Leu Lys Arg Arg Val Glu Asp Ala Val Phe Cys Leu Asn Thr Ile Glu305 310 315 320Asp Phe Pro Phe Arg Glu Pro Ile Arg Gln Pro Pro Asp Cys Ser Lys 325 330 335Val Leu Ile Glu Ala Met Glu Lys Tyr Phe Met Met Cys Ser Pro Lys 340 345 350Asp Arg Gln Ser Ala Ala Trp Leu Gly Ala Gly Val Val Glu Leu Ile 355 360 365Cys Asp Gly Asn Pro Leu Ser Glu Val Leu Gly Phe Leu Ala Lys Tyr 370 375 380Met Pro Ile Gln Lys Glu Cys Thr Gly Asn Leu Leu Lys Ile Tyr Ala385 390 395 400Leu Leu Thr Val202259DNAKaposi's sarcoma-associated herpesvirus 20atggcagcgc tcgagggccc cctactactg ccaccgagcg cctccctgac gacgagtccg 60cagaccacgt gttatcaagc gacttgggaa tcacagctgg aaatattctg ctgtctggcc 120accaactcgc acctgcaggc agagctgacc ttagaaggtc ttgataagat gatgcagccc 180gagcccacct ttttcgcctg cagagcgata cgcagactac tcctggggga acgcctccac 240ccttttatac atcaagaagg gactcttttg ggaaaagtgg gtcgacggta cagcggcgaa 300ggtttaataa ttgacggtgg tggagtgttt acgcgcggac agatagacac cgacaactac 360ctacctgcgg tgggatcatg ggaacttacc gatgattgtg ataaaccctg cgaattcagg 420gagctacgct cgctgtatct tcccgcgcta ctaacgtgca ccatatgtta caaagccatg 480ttcaggatag tgtgcaggta cctggagttc tgggagttcg aacagtgttt tcatgcgttt 540ctggcggtgt tgccccatag tctacaaccc acaatctatc aaaattattt tgcactcctg 600gagagcctga agcatctctc gttttcaata atgccacccg catccccaga cgcacagcta 660cattttttaa agtttaacat cagcagcttc atggccacgt gggggtggca cggagagctg 720gtctcgctgc gccgtgccat cgctcacaac gtagagcgac tgcccaccgt gctgaagaac 780ctgtcgaaac agagtaagca ccaggacgtc aaggttaacg gacgggatct ggtgggcttt 840cagctggctc taaaccagct cgtgtcccgt ctgcacgtaa aaatccaacg caaggacccc 900ggaccaaagc catacagggt ggtcgtcagt accccagatt gtacctacta tctagtgtat 960ccgggcacac cggccatcta cagactcgtc atgtgtatgg cagtggcaga ctgcatcggc 1020cactcgtgca gcggactgca cccctgcgca aactttttag gcacccacga gacaccgcgt 1080ctcctggcgg cgacgctttc aagaatccgg tacgcgccga aagaccggcg agcagccatg 1140aaaggaaatt tgcaggcgtg cttccaacga
tacgcggcca cggacgcgcg gactctgggc 1200agctctacag tgtcagacat gctggaaccc acaaaacacg tcagtttgga aaacttcaag 1260atcaccatat tcaacaccaa catggtgatt aacactaaga taagctgcca cgttcctaac 1320accctgcaaa agactatttt aaacatcccc agattgacca acaattttgt tatacgaaag 1380tactccgtaa aggaaccttc ttttaccata agcgtgtttt tttccgacaa catgtgtcaa 1440ggcaccgcaa taaacatcaa catcagtggg gacatgctgc actttctctt cgcaatgggt 1500acgctgaaat gctttctgcc aatcaggcac atatttcctg tatcgatagc aaattggaac 1560tccacgttgg acctgcacgg actggaaaac cagtacatgg tgagaatggg gcgaaaaaac 1620gtattttgga ccacaaactt tccatctgtg gtctccagca aggatgggct aaacgtgtcc 1680tggtttaagg ccgcgacagc cacgatttct aaagtgtacg ggcagcctct tgtggaacag 1740attcgccacg agctggcgcc cattctcacg gaccagcacg cgcgcatcga cggaaacaaa 1800aatagaatat tctccctact tgagcacaga aaccgttccc aaatacagac gctacacaaa 1860aggttcctgg agtgtctggt ggaatgctgt tcgtttctca ggcttgacgt ggcttgcatt 1920aggcgagccg ccgcccgggg cctgtttgac ttctcaaaga agataatcag tcacactaaa 1980agcaaacacg agtgcgcagt actgggatat aaaaagtgta acctaatccc gaaaatctat 2040gcccgaaaca agaagaccag gctagacgag ttgggccgca atgcaaactt catttcgttc 2100gtcgccacca cgggtcatcg gttcgccgct ctaaagccac aaattgtccg tcacgccatt 2160cgcaaactag gcctgcactg gcgccaccga acggccgcgt ccaacgagca gacaccgcca 2220gccgatcccc gcgtacgttg cgtccgtccg ctggtctaa 225921752PRTKaposi's sarcoma-associated herpesvirus 21Met Ala Ala Leu Glu Gly Pro Leu Leu Leu Pro Pro Ser Ala Ser Leu1 5 10 15Thr Thr Ser Pro Gln Thr Thr Cys Tyr Gln Ala Thr Trp Glu Ser Gln 20 25 30Leu Glu Ile Phe Cys Cys Leu Ala Thr Asn Ser His Leu Gln Ala Glu 35 40 45Leu Thr Leu Glu Gly Leu Asp Lys Met Met Gln Pro Glu Pro Thr Phe 50 55 60Phe Ala Cys Arg Ala Ile Arg Arg Leu Leu Leu Gly Glu Arg Leu His65 70 75 80Pro Phe Ile His Gln Glu Gly Thr Leu Leu Gly Lys Val Gly Arg Arg 85 90 95Tyr Ser Gly Glu Gly Leu Ile Ile Asp Gly Gly Gly Val Phe Thr Arg 100 105 110Gly Gln Ile Asp Thr Asp Asn Tyr Leu Pro Ala Val Gly Ser Trp Glu 115 120 125Leu Thr Asp Asp Cys Asp Lys Pro Cys Glu Phe Arg Glu Leu Arg Ser 130 135 140Leu Tyr Leu Pro Ala Leu Leu Thr Cys Thr Ile Cys Tyr Lys Ala Met145 150 155 160Phe Arg Ile Val Cys Arg Tyr Leu Glu Phe Trp Glu Phe Glu Gln Cys 165 170 175Phe His Ala Phe Leu Ala Val Leu Pro His Ser Leu Gln Pro Thr Ile 180 185 190Tyr Gln Asn Tyr Phe Ala Leu Leu Glu Ser Leu Lys His Leu Ser Phe 195 200 205Ser Ile Met Pro Pro Ala Ser Pro Asp Ala Gln Leu His Phe Leu Lys 210 215 220Phe Asn Ile Ser Ser Phe Met Ala Thr Trp Gly Trp His Gly Glu Leu225 230 235 240Val Ser Leu Arg Arg Ala Ile Ala His Asn Val Glu Arg Leu Pro Thr 245 250 255Val Leu Lys Asn Leu Ser Lys Gln Ser Lys His Gln Asp Val Lys Val 260 265 270Asn Gly Arg Asp Leu Val Gly Phe Gln Leu Ala Leu Asn Gln Leu Val 275 280 285Ser Arg Leu His Val Lys Ile Gln Arg Lys Asp Pro Gly Pro Lys Pro 290 295 300Tyr Arg Val Val Val Ser Thr Pro Asp Cys Thr Tyr Tyr Leu Val Tyr305 310 315 320Pro Gly Thr Pro Ala Ile Tyr Arg Leu Val Met Cys Met Ala Val Ala 325 330 335Asp Cys Ile Gly His Ser Cys Ser Gly Leu His Pro Cys Ala Asn Phe 340 345 350Leu Gly Thr His Glu Thr Pro Arg Leu Leu Ala Ala Thr Leu Ser Arg 355 360 365Ile Arg Tyr Ala Pro Lys Asp Arg Arg Ala Ala Met Lys Gly Asn Leu 370 375 380Gln Ala Cys Phe Gln Arg Tyr Ala Ala Thr Asp Ala Arg Thr Leu Gly385 390 395 400Ser Ser Thr Val Ser Asp Met Leu Glu Pro Thr Lys His Val Ser Leu 405 410 415Glu Asn Phe Lys Ile Thr Ile Phe Asn Thr Asn Met Val Ile Asn Thr 420 425 430Lys Ile Ser Cys His Val Pro Asn Thr Leu Gln Lys Thr Ile Leu Asn 435 440 445Ile Pro Arg Leu Thr Asn Asn Phe Val Ile Arg Lys Tyr Ser Val Lys 450 455 460Glu Pro Ser Phe Thr Ile Ser Val Phe Phe Ser Asp Asn Met Cys Gln465 470 475 480Gly Thr Ala Ile Asn Ile Asn Ile Ser Gly Asp Met Leu His Phe Leu 485 490 495Phe Ala Met Gly Thr Leu Lys Cys Phe Leu Pro Ile Arg His Ile Phe 500 505 510Pro Val Ser Ile Ala Asn Trp Asn Ser Thr Leu Asp Leu His Gly Leu 515 520 525Glu Asn Gln Tyr Met Val Arg Met Gly Arg Lys Asn Val Phe Trp Thr 530 535 540Thr Asn Phe Pro Ser Val Val Ser Ser Lys Asp Gly Leu Asn Val Ser545 550 555 560Trp Phe Lys Ala Ala Thr Ala Thr Ile Ser Lys Val Tyr Gly Gln Pro 565 570 575Leu Val Glu Gln Ile Arg His Glu Leu Ala Pro Ile Leu Thr Asp Gln 580 585 590His Ala Arg Ile Asp Gly Asn Lys Asn Arg Ile Phe Ser Leu Leu Glu 595 600 605His Arg Asn Arg Ser Gln Ile Gln Thr Leu His Lys Arg Phe Leu Glu 610 615 620Cys Leu Val Glu Cys Cys Ser Phe Leu Arg Leu Asp Val Ala Cys Ile625 630 635 640Arg Arg Ala Ala Ala Arg Gly Leu Phe Asp Phe Ser Lys Lys Ile Ile 645 650 655Ser His Thr Lys Ser Lys His Glu Cys Ala Val Leu Gly Tyr Lys Lys 660 665 670Cys Asn Leu Ile Pro Lys Ile Tyr Ala Arg Asn Lys Lys Thr Arg Leu 675 680 685Asp Glu Leu Gly Arg Asn Ala Asn Phe Ile Ser Phe Val Ala Thr Thr 690 695 700Gly His Arg Phe Ala Ala Leu Lys Pro Gln Ile Val Arg His Ala Ile705 710 715 720Arg Lys Leu Gly Leu His Trp Arg His Arg Thr Ala Ala Ser Asn Glu 725 730 735Gln Thr Pro Pro Ala Asp Pro Arg Val Arg Cys Val Arg Pro Leu Val 740 745 75022364DNAKaposi's sarcoma-associated herpesvirus 22atggtacgtc caaccgaggc cgaggttaag aaatccctga gcaggcttcc agcagcacgc 60aaaagagcag gtaaccgggc ccacctggcc acctaccgcc ggctcctcaa gtactccacc 120ctgcccgatc tatggcggtt tctaagtagc cggccccaga accctcccct tggacaccac 180agattattct ttgaggtgac tctagggcac agaattgccg actgcgtaat tctggtatcg 240ggtgggcatc agcccgtatg ttacgttgta gagctcaaga cttgtctgag tcaccagctg 300atcccaacca acaccgtgag aacgtcacag cgagctcaag gcctgtgcca actctccgac 360tcga 36423121PRTKaposi's sarcoma-associated herpesvirus 23Met Val Arg Pro Thr Glu Ala Glu Val Lys Lys Ser Leu Ser Arg Leu1 5 10 15Pro Ala Ala Arg Lys Arg Ala Gly Asn Arg Ala His Leu Ala Thr Tyr 20 25 30Arg Arg Leu Leu Lys Tyr Ser Thr Leu Pro Asp Leu Trp Arg Phe Leu 35 40 45Ser Ser Arg Pro Gln Asn Pro Pro Leu Gly His His Arg Leu Phe Phe 50 55 60Glu Val Thr Leu Gly His Arg Ile Ala Asp Cys Val Ile Leu Val Ser65 70 75 80Gly Gly His Gln Pro Val Cys Tyr Val Val Glu Leu Lys Thr Cys Leu 85 90 95Ser His Gln Leu Ile Pro Thr Asn Thr Val Arg Thr Ser Gln Arg Ala 100 105 110Gln Gly Leu Cys Gln Leu Ser Asp Ser 115 12024918DNAKaposi's sarcoma-associated herpesvirus 24atggcactcg acaagagtat agtggttaac ttcacctcca gactcttcgc tgatgaactg 60gccgcccttc agtcaaaaat agggagcgta ctgccgctcg gagattgcca ccgtttacaa 120aatatacagg cattgggcct ggggtgcgta tgctcacgtg agacatctcc ggactacatc 180caaattatgc agtatctatc caagtgcaca ctcgctgtcc tggaggaggt tcgcccggac 240agcctgcgcc taacgcggat ggatccctct gacaaccttc agataaaaaa cgtatatgcc 300cccttttttc agtgggacag caacacccag ctagcagtgc tacccccatt ttttagccga 360aaggattcca ccattgtgct cgaatccaac ggatttgacc ccgtgttccc catggtcgtg 420ccgcagcaac tggggcacgc tattctgcag cagctgttgg tgtaccacat ctactccaaa 480atatcggccg gggccccgga tgatgtaaat atggcggaac ttgatctata taccaccaat 540gtgtcattta tggggcgcac atatcgtctg gacgtagaca acacggatcc acgtactgcc 600ctgcgagtgc ttgacgatct gtccatgtac ctttgtatcc tatcagcctt ggttcccagg 660gggtgtctcc gtctgctcac ggcgctcgtg cggcacgaca ggcatcctct gacagaggtg 720tttgaggggg tggtgccaga tgaggtgacc aggatagatc tcgaccagtt gagcgtccca 780gatgacatca ccaggatgcg cgtcatgttc tcctatcttc agagtctcag ttctatattt 840aatcttggcc ccagactgca cgtgtatgcc tactcggcag agactttggc ggcctcctgt 900tggtattccc cacgctaa 91825305PRTKaposi's sarcoma-associated herpesvirus 25Met Ala Leu Asp Lys Ser Ile Val Val Asn Phe Thr Ser Arg Leu Phe1 5 10 15Ala Asp Glu Leu Ala Ala Leu Gln Ser Lys Ile Gly Ser Val Leu Pro 20 25 30Leu Gly Asp Cys His Arg Leu Gln Asn Ile Gln Ala Leu Gly Leu Gly 35 40 45Cys Val Cys Ser Arg Glu Thr Ser Pro Asp Tyr Ile Gln Ile Met Gln 50 55 60Tyr Leu Ser Lys Cys Thr Leu Ala Val Leu Glu Glu Val Arg Pro Asp65 70 75 80Ser Leu Arg Leu Thr Arg Met Asp Pro Ser Asp Asn Leu Gln Ile Lys 85 90 95Asn Val Tyr Ala Pro Phe Phe Gln Trp Asp Ser Asn Thr Gln Leu Ala 100 105 110Val Leu Pro Pro Phe Phe Ser Arg Lys Asp Ser Thr Ile Val Leu Glu 115 120 125Ser Asn Gly Phe Asp Pro Val Phe Pro Met Val Val Pro Gln Gln Leu 130 135 140Gly His Ala Ile Leu Gln Gln Leu Leu Val Tyr His Ile Tyr Ser Lys145 150 155 160Ile Ser Ala Gly Ala Pro Asp Asp Val Asn Met Ala Glu Leu Asp Leu 165 170 175Tyr Thr Thr Asn Val Ser Phe Met Gly Arg Thr Tyr Arg Leu Asp Val 180 185 190Asp Asn Thr Asp Pro Arg Thr Ala Leu Arg Val Leu Asp Asp Leu Ser 195 200 205Met Tyr Leu Cys Ile Leu Ser Ala Leu Val Pro Arg Gly Cys Leu Arg 210 215 220Leu Leu Thr Ala Leu Val Arg His Asp Arg His Pro Leu Thr Glu Val225 230 235 240Phe Glu Gly Val Val Pro Asp Glu Val Thr Arg Ile Asp Leu Asp Gln 245 250 255Leu Ser Val Pro Asp Asp Ile Thr Arg Met Arg Val Met Phe Ser Tyr 260 265 270Leu Gln Ser Leu Ser Ser Ile Phe Asn Leu Gly Pro Arg Leu His Val 275 280 285Tyr Ala Tyr Ser Ala Glu Thr Leu Ala Ala Ser Cys Trp Tyr Ser Pro 290 295 300Arg30526873DNAKaposi's sarcoma-associated herpesvirus 26atggcgtcat ctgatattct gtcggttgca aggacggatg acggctccgt ctgtgaagtc 60tccctgcgtg gaggtaggaa aaaaactacc gtctacctgc cggacactga accctgggtg 120gtagagaccg acgccatcaa agacgccttc ctcagcgacg ggatcgtgga tatggctcga 180aagcttcatc gtggtgccct gccctcaaat tctcacaacg gcttgaggat ggtgcttttt 240tgttattgtt acttgcaaaa ttgtgtgtac ctagccctgt ttctgtgccc ccttaatcct 300tacttggtaa ctccctcaag cattgagttt gccgagcccg ttgtggcacc tgaggtgctc 360ttcccacacc cggctgagat gtctcgcggt tgcgatgacg cgattttctg taaactgccc 420tataccgtgc ctataatcaa caccacgttt ggacgcattt acccgaactc tacacgcgag 480ccggacggca ggcctacgga ttactccatg gcccttagaa gggcttttgc agttatggtt 540aacacgtcat gtgcaggagt gacattgtgc cgcggagaaa ctcagaccgc atcccgtaac 600cacactgagt gggaaaatct gctggctatg ttttctgtga ttatctatgc cttagatcac 660aactgtcacc cggaagcact gtctatcgcg agcggcatct ttgacgagcg tgactatgga 720ttattcatct ctcagccccg gagcgtgccc tcgcctaccc cttgcgacgt gtcgtgggaa 780gatatctaca acgggactta cctagctcgg cctggaaact gtgacccctg gcccaatcta 840tccacccctc ccttgattct aaattttaaa taa 87327290PRTKaposi's sarcoma-associated herpesvirus 27Met Ala Ser Ser Asp Ile Leu Ser Val Ala Arg Thr Asp Asp Gly Ser1 5 10 15Val Cys Glu Val Ser Leu Arg Gly Gly Arg Lys Lys Thr Thr Val Tyr 20 25 30Leu Pro Asp Thr Glu Pro Trp Val Val Glu Thr Asp Ala Ile Lys Asp 35 40 45Ala Phe Leu Ser Asp Gly Ile Val Asp Met Ala Arg Lys Leu His Arg 50 55 60Gly Ala Leu Pro Ser Asn Ser His Asn Gly Leu Arg Met Val Leu Phe65 70 75 80Cys Tyr Cys Tyr Leu Gln Asn Cys Val Tyr Leu Ala Leu Phe Leu Cys 85 90 95Pro Leu Asn Pro Tyr Leu Val Thr Pro Ser Ser Ile Glu Phe Ala Glu 100 105 110Pro Val Val Ala Pro Glu Val Leu Phe Pro His Pro Ala Glu Met Ser 115 120 125Arg Gly Cys Asp Asp Ala Ile Phe Cys Lys Leu Pro Tyr Thr Val Pro 130 135 140Ile Ile Asn Thr Thr Phe Gly Arg Ile Tyr Pro Asn Ser Thr Arg Glu145 150 155 160Pro Asp Gly Arg Pro Thr Asp Tyr Ser Met Ala Leu Arg Arg Ala Phe 165 170 175Ala Val Met Val Asn Thr Ser Cys Ala Gly Val Thr Leu Cys Arg Gly 180 185 190Glu Thr Gln Thr Ala Ser Arg Asn His Thr Glu Trp Glu Asn Leu Leu 195 200 205Ala Met Phe Ser Val Ile Ile Tyr Ala Leu Asp His Asn Cys His Pro 210 215 220Glu Ala Leu Ser Ile Ala Ser Gly Ile Phe Asp Glu Arg Asp Tyr Gly225 230 235 240Leu Phe Ile Ser Gln Pro Arg Ser Val Pro Ser Pro Thr Pro Cys Asp 245 250 255Val Ser Trp Glu Asp Ile Tyr Asn Gly Thr Tyr Leu Ala Arg Pro Gly 260 265 270Asn Cys Asp Pro Trp Pro Asn Leu Ser Thr Pro Pro Leu Ile Leu Asn 275 280 285Phe Lys 29028363DNAKaposi's sarcoma-associated herpesvirus 28atgagcatga ctttccccgt ctccagtcac cggaggaatg gtggacggct ccgtcctggt 60gcgaatggcc accaagcctc ccgtgattgg tcttataaca gtgctcttcc tcctagtcat 120aggcgcctgc gtctactgct gcattcgcgt gttcctggcg gctcgactgt ggcgcgccac 180cccactaggc agggccaccg tggcgtatca ggtccttcgc accctgggac cgcaggccgg 240gtcacatgca ccgccgacgg tgggcatagc tacccaggag ccctaccgta caatatacat 300gccagattag aacggggtgt gtgctataat ggatggctat gggggggggc tgtagataat 360tga 36329120PRTKaposi's sarcoma-associated herpesvirus 29Met Ser Met Thr Phe Pro Val Ser Ser His Arg Arg Asn Gly Gly Arg1 5 10 15Leu Arg Pro Gly Ala Asn Gly His Gln Ala Ser Arg Asp Trp Ser Tyr 20 25 30Asn Ser Ala Leu Pro Pro Ser His Arg Arg Leu Arg Leu Leu Leu His 35 40 45Ser Arg Val Pro Gly Gly Ser Thr Val Ala Arg His Pro Thr Arg Gln 50 55 60Gly His Arg Gly Val Ser Gly Pro Ser His Pro Gly Thr Ala Gly Arg65 70 75 80Val Thr Cys Thr Ala Asp Gly Gly His Ser Tyr Pro Gly Ala Leu Pro 85 90 95Tyr Asn Ile His Ala Arg Leu Glu Arg Gly Val Cys Tyr Asn Gly Trp 100 105 110Leu Trp Gly Gly Ala Val Asp Asn 115 12030921DNAKaposi's sarcoma-associated herpesvirus 30atgctgctca gccgtcacag ggagcgcctt gccgccaacc tggaggagac cgccaaagac 60gccggagaga ggtgggaact gagtgccccg acattcacgc gacactgtcc caaaacggca 120cggatggcgc acccttttat tggcgtggtg cacagaataa actcatacag ttcggtcctg 180gaaacatact gcacacggca ccatcccgcc acgcccacgt cagcaaatcc cgacgtggga 240acccccagac cgtccgagga caacgtcccc gcaaagccgc gcctattgga gtccctatca 300acatacttgc agatgcggtg tgtgcgcgag gacgcgcacg tctccacggc cgatcaactg 360gtcgagtacc aggcgggcag aaaaacacac gactccctgc acgcctgctc tgtctaccgc 420gaacttcagg cttttctggt taacctttcg tcctttctga acggctgtta cgttcccggg 480gtgcactggc tggagccctt ccaacagcag ctagtaatgc acactttttt ctttttggtt 540tcaatcaagg ccccacaaaa gacgcaccag ttgtttggat tgtttaagca gtacttcggt 600ttatttgaaa ctccaaacag tgttttacag acgtttaagc aaaaggcaag cgtattccta 660ataccaagga gacacggaaa gacatggata gtggtggcga tcatcagcat gctactggca 720tccgtagaga acattaacat tgggtacgta gcccaccaaa agcacgtagc caactccgtg 780ttcgcggaaa tcataaagac gctttgtcgg tggttccccc ccaaaaattt aaacatcaag 840aaggagaacg gaaccataat ctacacgcga cccggaggac ggtccagctc gctgatgtgc 900gcaacatgct tcaataagaa c 92131307PRTKaposi's sarcoma-associated herpesvirus 31Met Leu Leu Ser Arg His Arg Glu Arg Leu Ala Ala Asn Leu Glu Glu1 5 10 15Thr Ala Lys Asp Ala Gly Glu Arg Trp Glu Leu
Ser Ala Pro Thr Phe 20 25 30Thr Arg His Cys Pro Lys Thr Ala Arg Met Ala His Pro Phe Ile Gly 35 40 45Val Val His Arg Ile Asn Ser Tyr Ser Ser Val Leu Glu Thr Tyr Cys 50 55 60Thr Arg His His Pro Ala Thr Pro Thr Ser Ala Asn Pro Asp Val Gly65 70 75 80Thr Pro Arg Pro Ser Glu Asp Asn Val Pro Ala Lys Pro Arg Leu Leu 85 90 95Glu Ser Leu Ser Thr Tyr Leu Gln Met Arg Cys Val Arg Glu Asp Ala 100 105 110His Val Ser Thr Ala Asp Gln Leu Val Glu Tyr Gln Ala Gly Arg Lys 115 120 125Thr His Asp Ser Leu His Ala Cys Ser Val Tyr Arg Glu Leu Gln Ala 130 135 140Phe Leu Val Asn Leu Ser Ser Phe Leu Asn Gly Cys Tyr Val Pro Gly145 150 155 160Val His Trp Leu Glu Pro Phe Gln Gln Gln Leu Val Met His Thr Phe 165 170 175Phe Phe Leu Val Ser Ile Lys Ala Pro Gln Lys Thr His Gln Leu Phe 180 185 190Gly Leu Phe Lys Gln Tyr Phe Gly Leu Phe Glu Thr Pro Asn Ser Val 195 200 205Leu Gln Thr Phe Lys Gln Lys Ala Ser Val Phe Leu Ile Pro Arg Arg 210 215 220His Gly Lys Thr Trp Ile Val Val Ala Ile Ile Ser Met Leu Leu Ala225 230 235 240Ser Val Glu Asn Ile Asn Ile Gly Tyr Val Ala His Gln Lys His Val 245 250 255Ala Asn Ser Val Phe Ala Glu Ile Ile Lys Thr Leu Cys Arg Trp Phe 260 265 270Pro Pro Lys Asn Leu Asn Ile Lys Lys Glu Asn Gly Thr Ile Ile Tyr 275 280 285Thr Arg Pro Gly Gly Arg Ser Ser Ser Leu Met Cys Ala Thr Cys Phe 290 295 300Asn Lys Asn305321365DNAKaposi's sarcoma-associated herpesvirus 32atggatgcgc atgctatcaa cgaaagatac gtaggtcctc gctgccaccg tttggcccac 60gtggtgctgc ctaggacctt tctgctgcat cacgccatac ccctggagcc cgagatcatc 120ttttccacct acacccggtt cagccggtcg ccagggtcat cccgccggtt ggtggtgtgt 180gggaaacgtg tcctgccagg ggaggaaaac caacttgcgt cttcaccttc tggtttggcg 240cttagcctgc ctctgttttc ccacgatggg aactttcatc catttgacat ctcggtactg 300cgcatttcct gccctggttc taatcttagt cttactgtca gatttctcta tctatctctg 360gtggtggcta tgggggcggg acggaataat gcgcggagtc cgaccgttga cggggtatcg 420ccgccagagg gcgccgtagc ccaccctttg gaggaactgc agaggctggc gcgtgctacg 480ccggacccgg cactcacccg tggaccgttg caggtcctga ccggccttct ccgcgcaggg 540tcagacggag accgcgccac tcaccacatg gcgctcgagg ctccgggaac cgtgcgtgga 600gaaagcctag acccgcctgt ttcacagaag gggccagcgc gcacacgcca caggccaccc 660cccgtgcgac tgagcttcaa ccccgtcaat gccgatgtac ccgctacctg gcgagacgcc 720actaacgtgt actcgggtgc tccctactat gtgtgtgttt acgaacgcgg tggccgtcag 780gaagacgact ggctgccgat accactgagc ttcccagaag agcccgtgcc cccgccaccg 840ggcttagtgt tcatggacga cttgttcatt aacacgaagc agtgcgactt tgtggacacg 900ctagaggccg cctgtcgcac gcaaggctac acgttgagac agcgcgtgcc tgtcgccatt 960cctcgcgacg cggaaatcgc agacgcagtt aaatcgcact ttttagaggc gtgcctagtg 1020ttacgggggc tggcttcgga ggctagtgcc tggataagag ctgccacgtc cccgcccctt 1080ggccgccacg cctgctggat ggacgtgtta ggattatggg aaagccgccc ccacactcta 1140ggtttggagt tacgcggcgt aaactgtggc ggcacggacg gtgactggtt agagatttta 1200aaacagcccg atgtgcaaaa gacagtcagc gggagtcttg tggcatgcgt gatcgtcaca 1260cccgcattgg aagcctggct tgtgttacct gggggttttg ctattaaagc ccgctatagg 1320gcgtcgaagg aggatctggt gttcattcga ggccgctatg gctag 136533454PRTKaposi's sarcoma-associated herpesvirus 33Met Asp Ala His Ala Ile Asn Glu Arg Tyr Val Gly Pro Arg Cys His1 5 10 15Arg Leu Ala His Val Val Leu Pro Arg Thr Phe Leu Leu His His Ala 20 25 30Ile Pro Leu Glu Pro Glu Ile Ile Phe Ser Thr Tyr Thr Arg Phe Ser 35 40 45Arg Ser Pro Gly Ser Ser Arg Arg Leu Val Val Cys Gly Lys Arg Val 50 55 60Leu Pro Gly Glu Glu Asn Gln Leu Ala Ser Ser Pro Ser Gly Leu Ala65 70 75 80Leu Ser Leu Pro Leu Phe Ser His Asp Gly Asn Phe His Pro Phe Asp 85 90 95Ile Ser Val Leu Arg Ile Ser Cys Pro Gly Ser Asn Leu Ser Leu Thr 100 105 110Val Arg Phe Leu Tyr Leu Ser Leu Val Val Ala Met Gly Ala Gly Arg 115 120 125Asn Asn Ala Arg Ser Pro Thr Val Asp Gly Val Ser Pro Pro Glu Gly 130 135 140Ala Val Ala His Pro Leu Glu Glu Leu Gln Arg Leu Ala Arg Ala Thr145 150 155 160Pro Asp Pro Ala Leu Thr Arg Gly Pro Leu Gln Val Leu Thr Gly Leu 165 170 175Leu Arg Ala Gly Ser Asp Gly Asp Arg Ala Thr His His Met Ala Leu 180 185 190Glu Ala Pro Gly Thr Val Arg Gly Glu Ser Leu Asp Pro Pro Val Ser 195 200 205Gln Lys Gly Pro Ala Arg Thr Arg His Arg Pro Pro Pro Val Arg Leu 210 215 220Ser Phe Asn Pro Val Asn Ala Asp Val Pro Ala Thr Trp Arg Asp Ala225 230 235 240Thr Asn Val Tyr Ser Gly Ala Pro Tyr Tyr Val Cys Val Tyr Glu Arg 245 250 255Gly Gly Arg Gln Glu Asp Asp Trp Leu Pro Ile Pro Leu Ser Phe Pro 260 265 270Glu Glu Pro Val Pro Pro Pro Pro Gly Leu Val Phe Met Asp Asp Leu 275 280 285Phe Ile Asn Thr Lys Gln Cys Asp Phe Val Asp Thr Leu Glu Ala Ala 290 295 300Cys Arg Thr Gln Gly Tyr Thr Leu Arg Gln Arg Val Pro Val Ala Ile305 310 315 320Pro Arg Asp Ala Glu Ile Ala Asp Ala Val Lys Ser His Phe Leu Glu 325 330 335Ala Cys Leu Val Leu Arg Gly Leu Ala Ser Glu Ala Ser Ala Trp Ile 340 345 350Arg Ala Ala Thr Ser Pro Pro Leu Gly Arg His Ala Cys Trp Met Asp 355 360 365Val Leu Gly Leu Trp Glu Ser Arg Pro His Thr Leu Gly Leu Glu Leu 370 375 380Arg Gly Val Asn Cys Gly Gly Thr Asp Gly Asp Trp Leu Glu Ile Leu385 390 395 400Lys Gln Pro Asp Val Gln Lys Thr Val Ser Gly Ser Leu Val Ala Cys 405 410 415Val Ile Val Thr Pro Ala Leu Glu Ala Trp Leu Val Leu Pro Gly Gly 420 425 430Phe Ala Ile Lys Ala Arg Tyr Arg Ala Ser Lys Glu Asp Leu Val Phe 435 440 445Ile Arg Gly Arg Tyr Gly 45034984DNAKaposi's sarcoma-associated herpesvirus 34atgtttgctt tgagctcgct cgtgtccgag ggtgacccgg aggtgaccag taggtacgtc 60aagggcgtac aacttgccct ggaccttagc gagaacacac ctggacaatt taagttgata 120gaaactcccc tgaacagctt cctcttggtt tccaacgtga tgcccgaggt ccagccaatc 180tgcagtggcc ggccggcctt gcggccagac tttagtaatc tccacttgcc tagactggag 240aagctccaga gagtcctcgg gcagggtttc ggggcggcgg gtgaggaaat cgcactggac 300ccgtctcacg tagaaacaca cgaaaagggc caggtgttct acaaccacta tgctaccgag 360gagtggacgt gggctttgac tctgaataag gatgcgctcc ttcgggaggc tgtagatggc 420ctgtgtgacc ccggaacttg gaagggtctt cttcctgacg acccccttcc gttgctatgg 480ctgctgttca acggacccgc ctctttttgt cgggccgact gttgcctgta caagcagcac 540tgcggttacc cgggcccggt gctacttcca ggtcacatgt acgctcccaa acgggatctt 600ttgtcgttcg ttaatcatgc cctgaagtac accaagtttc tatacggaga tttttccggg 660acatgggcgg cggcttgccg cccgccattc gctacttctc ggatacaaag ggtagtgagt 720cagatgaaaa tcatagatgc ttccgacact tacatttccc acacctgcct cttgtgtcac 780atatatcagc aaaatagcat aattgcgggt caggggaccc acgtgggtgg aatcctactg 840ttgagtggaa aagggaccca gtatataaca ggcaatgttc agacccaaag gtgtccaact 900acgggcgact atctaatcat cccatcgtat gacataccgg cgatcatcac catgatcaag 960gagaatggac tcaaccaact ctaa 98435327PRTKaposi's sarcoma-associated herpesvirus 35Met Phe Ala Leu Ser Ser Leu Val Ser Glu Gly Asp Pro Glu Val Thr1 5 10 15Ser Arg Tyr Val Lys Gly Val Gln Leu Ala Leu Asp Leu Ser Glu Asn 20 25 30Thr Pro Gly Gln Phe Lys Leu Ile Glu Thr Pro Leu Asn Ser Phe Leu 35 40 45Leu Val Ser Asn Val Met Pro Glu Val Gln Pro Ile Cys Ser Gly Arg 50 55 60Pro Ala Leu Arg Pro Asp Phe Ser Asn Leu His Leu Pro Arg Leu Glu65 70 75 80Lys Leu Gln Arg Val Leu Gly Gln Gly Phe Gly Ala Ala Gly Glu Glu 85 90 95Ile Ala Leu Asp Pro Ser His Val Glu Thr His Glu Lys Gly Gln Val 100 105 110Phe Tyr Asn His Tyr Ala Thr Glu Glu Trp Thr Trp Ala Leu Thr Leu 115 120 125Asn Lys Asp Ala Leu Leu Arg Glu Ala Val Asp Gly Leu Cys Asp Pro 130 135 140Gly Thr Trp Lys Gly Leu Leu Pro Asp Asp Pro Leu Pro Leu Leu Trp145 150 155 160Leu Leu Phe Asn Gly Pro Ala Ser Phe Cys Arg Ala Asp Cys Cys Leu 165 170 175Tyr Lys Gln His Cys Gly Tyr Pro Gly Pro Val Leu Leu Pro Gly His 180 185 190Met Tyr Ala Pro Lys Arg Asp Leu Leu Ser Phe Val Asn His Ala Leu 195 200 205Lys Tyr Thr Lys Phe Leu Tyr Gly Asp Phe Ser Gly Thr Trp Ala Ala 210 215 220Ala Cys Arg Pro Pro Phe Ala Thr Ser Arg Ile Gln Arg Val Val Ser225 230 235 240Gln Met Lys Ile Ile Asp Ala Ser Asp Thr Tyr Ile Ser His Thr Cys 245 250 255Leu Leu Cys His Ile Tyr Gln Gln Asn Ser Ile Ile Ala Gly Gln Gly 260 265 270Thr His Val Gly Gly Ile Leu Leu Leu Ser Gly Lys Gly Thr Gln Tyr 275 280 285Ile Thr Gly Asn Val Gln Thr Gln Arg Cys Pro Thr Thr Gly Asp Tyr 290 295 300Leu Ile Ile Pro Ser Tyr Asp Ile Pro Ala Ile Ile Thr Met Ile Lys305 310 315 320Glu Asn Gly Leu Asn Gln Leu 32536330DNAKaposi's sarcoma-associated herpesvirus 36ggatccctct gacaaccttc agataaaaaa cgtatatgcc cccttttttc agtgggacag 60caacacccag ctagcagtgc tacccccatt ttttagccga aaggattcca ccattgtgct 120cgaatccaac ggatttgacc ccgtgttccc catggtcgtg ccgcagcaac tggggcacgc 180tattctgcag cagctgttgg tgtaccacat ctactccaaa atatcggccg gggccccgga 240tgatgtaaat atggcggaac ttgatctata taccaccaat gtgtcattta tggggcgcac 300atatcgtctg gacgtagaca acacggatcc 33037627DNAKaposi's sarcoma-associated herpesvirus 37ggatccgctg gcaggtgggc gcgcacctcg tcgggtagct tggagacaaa cagctccagg 60ccagtccgcg ccgtagcgcc tgcaggtgcc tcaccaccgg ggccgggtca tgcgatctgt 120ttagtccgga gaagataggg cccttgggaa gccgctgaac cagctccagg gtctccaaga 180tgcgcaccgg ttgtcggagc tgtcgcgata gaggttaggg taggtgtccg gtccgtccgt 240gggctcaaac ctgcccagac acaccactgt ctgctggggg atcatccttc tcagggagat 300gcattctttg gaagtagtgg tagagatgga gcagactgcc agggcgttgc aggagtggtg 360gcgatggtgc gcaccgtttt taagaaaccc cccagggtgg ggactcccgc tccctgcagc 420atctcggcct gctgtacgtc cttggcgaat atgcgacgaa atcggctgtg cgcacggggt 480cccagggccg gtccggtggc atacaggccg gtgagggccc cctgggtctg tccgcctgga 540aacagggtgc tgtgaaacaa caggttgcaa ggccgcgaat acccctctgc acgctgctgt 600ggacgtgggt gtatgctccg tggatcc 62738233DNAArtificial SequenceProbe 38agccgaaagg attccaccat tgtgctcgaa tccaacggat ttgaccccgt gttccccatg 60gtcgtgccgc agcaactggg gcacgctatt ctgcagcagc tgttggtgta ccacatctac 120tccaaaatat cggccggggc cccggatgat gtaaatatgg cggaacttga tctatatacc 180accaatgtgt catttatggg gcgcacatat cgtctggacg tagacaacac gga 23339328DNAArtificial SequenceProbe 39gaaattaccc acgagatcgc ttccctgcac accgcacttg gctactcatc agtcatcgcc 60ccggcccacg tggccgccat aactacagac atgggagtac attgtcagga cctctttatg 120attttcccag gggacgcgta tcaggaccgc cagctgcatg actatatcaa aatgaaagcg 180ggcgtgcaaa ccggctcacc gggaaacaga atggatcacg tgggatacac tgctggggtt 240cctcgctgcg agaacctgcc cggtttgagt catggtcagc tggcaacctg cgagataatt 300cccacgccgg tcacatctga cgttgcct 32840132DNAArtificial SequenceProbe 40aacacgtcat gtgcaggagt gacattgtgc cgcggagaaa ctcagaccgc atcccgtaac 60cacactgagt gggaaaatct gctggctatg ttttctgtga ttatctatgc cttagatcac 120aactgtcacc cg 1324120DNAArtificial SequencePrimer 41agccgaaagg attccaccat 204220DNAArtificial SequencePrimer 42gaaattaccc acgagatcgc 204323DNAArtificial SequencePrimer 43aacacgtcat gtgcaggagt gac 234421DNAArtificial SequencePrimer 44acagggctgg ttgcccaggg t 214520DNAArtificial SequencePrimer 45agttgcaaac cagacctcag 2046304PRTKaposi's sarcoma-associated herpesvirus 46Met Leu Thr Asp Lys Thr Ile Ile Val Ser Leu Thr Ser Arg Leu Phe1 5 10 15Ala Asp Glu Ile Thr Lys Leu Gln Lys Lys Ile Gly Ser Ile Leu Pro 20 25 30Leu Gln Asp Pro His Lys Leu Gln Ser Leu Asp Thr Leu Gly Leu Asn 35 40 45Ala Val Cys Ser Arg Asp Val Phe Pro Asp Tyr Val His Met Phe Ser 50 55 60Tyr Leu Ser Lys Cys Thr Leu Ala Ile Leu Glu Glu Val Asn Pro Asp65 70 75 80Asn Leu Ile Leu Thr Arg Leu Asp Pro Ser Glu Thr Tyr Gln Ile Lys 85 90 95Asn Val Tyr Glu Pro Met Phe Gln Trp Asp Gly Phe Ser Asn Leu Thr 100 105 110Val Ile Pro Pro Val Phe Gly Arg Gln Gln Ala Thr Val Thr Leu Glu 115 120 125Ser Asn Gly Phe Asp Leu Val Phe Pro Ser Val Val Pro Ser Asp Leu 130 135 140Ala Gln Ala Ile Ile Gly Lys Leu Leu Leu Tyr Asn Leu Tyr Ser Arg145 150 155 160Leu Val Glu Ser Asp Pro Glu Ile Asn Ile Glu Glu Val Asn Met Tyr 165 170 175Thr Thr Asn Val Thr His Met Gly Arg His Tyr Val Leu Asp Ile Asn 180 185 190His Asn Asn Pro Asn Glu Ala Leu Lys Ser Leu Asp Asp Leu Ala Val 195 200 205Tyr Thr Lys Ile Leu Ser Ala Leu Ile Pro Arg Ala Lys Leu Arg Val 210 215 220Leu Thr Ile Leu Met Arg His Asp Gln His Glu Leu Leu Asp Val Phe225 230 235 240Arg Gly Ile Val Pro Arg Glu Val Tyr Glu Ile Asp Ala Asn Ala Leu 245 250 255Ser Ile Gly Asp Asp Ile Thr Arg Met Thr Thr Phe Ile Thr Tyr Leu 260 265 270Gln Ser Leu Ser Ser Ile Phe Asn Leu Gly Ala Lys Leu His Leu Ser 275 280 285Ser Tyr Ala Ser Glu Thr Gln Thr Ala Thr Cys Trp Ile Ser Tyr Cys 290 295 30047301PRTEpstein Barr Virus 47Met Asp Leu Lys Val Val Val Ser Leu Ser Ser Arg Leu Tyr Thr Asp1 5 10 15Glu Ile Ala Lys Met Gln Gln Arg Ile Gly Cys Ile Leu Pro Leu Ala 20 25 30Ser Thr His Gly Thr Gln Asn Val Gln Gly Leu Gly Leu Gly Gln Val 35 40 45Tyr Ser Leu Glu Thr Val Pro Asp Tyr Val Ser Met Tyr Asn Tyr Leu 50 55 60Ser Asp Cys Thr Leu Ala Val Leu Asp Glu Val Ser Val Asp Ser Leu65 70 75 80Ile Leu Thr Lys Ile Val Pro Gly Gln Thr Tyr Ala Ile Lys Asn Lys 85 90 95Tyr Gln Pro Phe Phe Gln Trp His Gly Thr Gly Ser Lys Ser Val Met 100 105 110Pro Pro Val Phe Gly Arg Glu His Ala Thr Val Lys Leu Glu Ser Asn 115 120 125Asp Val Asp Ile Val Phe Pro Met Val Leu Pro Thr Pro Ile Ala Glu 130 135 140Glu Val Leu Gln Lys Ile Leu Leu Phe Asn Val Tyr Ser Arg Val Val145 150 155 160Met Gln Ala Pro Gly Asn Ala Asp Met Leu Asp Val His Met His Leu 165 170 175Gly Ser Val Ser Tyr Leu Gly His His Tyr Glu Leu Ala Leu Pro Glu 180 185 190Val Pro Gly Pro Leu Gly Leu Ala Leu Leu Asp Asn Leu Ser Leu Tyr 195 200 205Phe Cys Ile Met Val Thr Leu Leu Pro Arg Ala Ser Met Arg Leu Val 210 215 220Arg Gly Leu Ile Arg His Glu His His Asp Leu Leu Asn Leu Phe Gln225 230 235 240Glu Met Val Pro Asp Glu Ile Ala Arg Ile Arg Leu Asp Asp Leu Ser 245 250 255Val Ala Asp Asp Leu Ser Arg Met Arg Val Met Met Thr Tyr Leu Gln 260 265 270Ser Leu Ala Ser Leu Phe Asn Leu Gly Pro Arg Leu
Ala Thr Ala Ala 275 280 285Tyr Ser Gln Glu Thr Leu Thr Ala Thr Cys Trp Leu Arg 290 295 3004820DNAArtificial SequencePrimer 48tccgtgttgt ctacgtccag 204918DNAArtificial SequencePrimer 49aggcaacgtc agatgtga 185023DNAArtificial SequencePrimer 50cgggtgacag ttgtgatcta agg 235120DNAArtificial SequencePrimer 51agcactcgca gggcagtacg 205222DNAArtificial SequencePrimer 52gactcttcgc tgatgaaact gg 225320DNAArtificial SequencePrimer 53tccgtgttgt ctacgtccag 205419DNAArtificial SequencePrimer 54aggcaacgtc agatgtgac 195525DNAArtificial SequencePrimer 55catgggagta cattgtcagg acctc 255621DNAArtificial SequencePrimer 56ggaattatct cgcaggttgc c 215723DNAArtificial SequencePrimer 57ggcgacattc atcaacctca ggg 235823DNAArtificial SequencePrimer 58atatcatcct gtgcgttcac gac 23
Patent applications by Patrick S. Moore, New York, NY US
Patent applications by Yuan Chang, New York, NY US
Patent applications in class Transgenic nonhuman animal (e.g., mollusks, etc.)
Patent applications in all subclasses Transgenic nonhuman animal (e.g., mollusks, etc.)