Patent application title: Brittle Stalk 2 Gene Family And Related Methods And Uses
Inventors:
Kanwarpal Singh Dhugga (Johnston, IA, US)
IPC8 Class: AA01H100FI
USPC Class:
800278
Class name: Multicellular living organisms and unmodified parts thereof and related processes method of introducing a polynucleotide molecule into or rearrangement of genetic material within a plant or plant part
Publication date: 2009-03-26
Patent application number: 20090083879
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Patent application title: Brittle Stalk 2 Gene Family And Related Methods And Uses
Inventors:
Kanwarpal Singh Dhugga
Agents:
E I DU PONT DE NEMOURS AND COMPANY;LEGAL PATENT RECORDS CENTER
Assignees:
Origin: WILMINGTON, DE US
IPC8 Class: AA01H100FI
USPC Class:
800278
Abstract:
This invention relates to isolated polynucleotides encoding BRITTLE STALK
2-like (Bk2L) family polypeptides. The invention also relates to the
construction of a chimeric gene encoding all or a portion of a Bk2L
polypeptide, in sense or antisense orientation, wherein expression of the
chimeric gene results in production of altered levels of the Bk2L
polypeptide in a transformed host cell.Claims:
1-21. (canceled)
22. An isolated polynucleotide comprising:(a) a nucleotide sequence encoding a polypeptide that increases stalk mechanical strength of a transformed plant expressing said polypeptide when compared to an untransformed control plant, wherein said polypeptide has an amino acid sequence of at least 96% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:6; or(b) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary.
23. A recombinant DNA construct comprising the polynucleotide of claim 22 operably linked to a promoter that is functional in a plant.
24. A plant comprising in its genome the recombinant DNA construct of claim 23.
25. The isolated polynucleotide of claim 22, wherein said stalk mechanical strength is measured by the three-point bend test.
26. The isolated polynucleotide of claim 22, wherein the polypeptide comprises SEQ ID NO:6.
Description:
FIELD OF THE INVENTION
[0001]The field of invention relates to plant molecular biology, and in particular, to BRITTLE STALK 2-like genes, BRITTLE STALK 2-like polypeptides, and uses thereof.
BACKGROUND OF THE INVENTION
[0002]Plant primary growth is mainly driven by an enlargement of the cells, which occurs through the irreversible yielding of the primary cell wall to turgor pressure inside the cell. Although cell division is required to produce new cells, the growth results from the expansion of these cells, not simply from their division. Cellulose microfibrils, which are embedded in a matrix of hemicellulose and lignin in the wall, are the main determinants of tensile strength (Appenzeller et al., Cellulose 11:287-299 (2004)). A cell usually expands along the axis that is perpendicular to the orientation of the microfibrils. For example, radial deposition of microfibrils favors cell expansion along the longitudinal axis.
[0003]Secondary wall differs from primary wall in that it is richer in cellulose and lignin and its deposition commences toward the end of cell expansion. Modulation of primary cell wall synthesis has applications in altering growth rate and size (stature) of a plant whereas that of secondary wall can be useful in improving biomass accumulation and tissue strength (Appenzeller et al., Cellulose 11:287-299 (2004)).
[0004]Cellulose in general is the major wall constituent in mature plant cells forming vegetative tissues. The paracrystalline structure of cellulose that results from energy minimization by the formation of inter- and intra-chain hydrogen bonds makes it mechanically one of the strongest organic molecule known on density basis. It is natural then that cellulose is the primary determinant of strength in structural tissues.
[0005]Plant mechanical strength is one of the most important agronomic traits. Plant mutants that are defective in stem strength have been isolated and characterized. Barley brittle culm (bc) mutants were first described based on the physical properties of the culms, which have an 80% reduction in the amount of cellulose and a twofold decrease in breaking strength compared with those of wildtype plants (Kokubo et al., Plant Physiol. 97:509-514 (1991)). Rice brittle culm1 (bc1) mutants show a reduction in cell wall thickness and cellulose content (Qian et al., Chi. Sci. Bull. 46:2082-2085 (2001)). Li et al. described the identification of rice BRITTLE CULM1 (BC1), a gene that encodes a COBRA-like protein (The Plant Cell 15(9):2020-2031 (2003)). Their findings indicated that BC1 functions in regulating the biosynthesis of secondary cell walls to provide the main mechanical strength for rice plants.
[0006]The stalks of maize brittle stalk 2 (bk2) mutant exhibit a dramatically reduced mechanical strength compared to their wild type counterparts (Langham, MNL 14:21-22 (1940)). Maize bk2 mutants have stalk and leaves that are very brittle and break easily. The main chemical constituent deficient in the mutant stalk is cellulose. Therefore, stalk mechanical strength appears to be dependent primarily on the amount of cellulose in a unit length of the stalk below the ear.
[0007]Furthermore, genes encoding cellulose synthase catalytic subunits (CesA) have been implicated in cell wall synthesis and are represented by a large family in plants. Ten genes were identified in Arabidopsis after complete genome sequencing and twelve genes have been isolated from maize by EST sequencing (U.S. Pat. Nos. 6,803,498 and 6,930,225). Three of the CesA genes from each Arabidopsis and maize have been reported to make secondary wall whereas the rest apparently make primary wall (Taylor et al., Proc. Natl. Acad. Sci. U.S.A. 100:1450-1455 (2003)). Mutations in three of the CesA genes from Arabidopsis resulted in collapsed xylem and reduced mechanical strength of the stem-like peduncle. When related CesA genes from rice were mutated the culms became brittle, indicating the role of these genes in secondary wall formation. In each case, reduced mechanical strength was correlated with diminished cellulose content.
[0008]In general, mutations in the CesA genes involved in primary wall formation cause severe phenotypic alterations whereas those in secondary wall-forming genes do not alter the visual phenotype as much as they affect mechanical strength (Appenzeller et al., Cellulose 11:287-299 (2004)).
[0009]As insufficient stalk strength is a major problem in corn breeding, it is desirable to provide compositions and methods for manipulating cellulose concentration in the cell wall and thereby alter plant stalk strength and/or quality for improved standability or silage quality.
SUMMARY OF THE INVENTION
The Present Invention Includes
[0010]In one embodiment, an isolated polynucleotide comprising (a) a nucleotide sequence encoding a polypeptide associated with stalk mechanical strength, wherein said polypeptide has an amino acid sequence of at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity, or any integer in between 80% and 100%, based on the Clustal V method of alignment, when compared to SEQ ID NOs:16 or 18, or (b) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary.
[0011]In another embodiment, a method of altering (preferably increasing) stalk mechanical strength in a plant comprising (a) introducing into a regenerable plant cell a recombinant DNA construct to produce transformed plant cells, said recombinant DNA construct comprising a promoter that is functional in a plant operably linked to (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:4, 6, 8, 10, 12, 14, 16, and 18, or (b) a full-length complement of said polynucleotide of (a)(i); and (b) regenerating a transgenic plant from said transformed plant cell, wherein said transgenic plant comprises in its genome said recombinant DNA construct and wherein said transgenic plant exhibits an alteration (preferably an increase) in stalk mechanical strength, when compared to a control plant not comprising said recombinant DNA construct. The method may further comprise (c) obtaining a progeny plant derived from said transgenic plant, wherein said progeny plant comprises in its genome the recombinant DNA construct.
[0012]In another embodiment, a method of evaluating stalk mechanical strength in a plant comprising (a) introducing into a regenerable plant cell a recombinant DNA construct to produce transformed plant cells, said recombinant DNA construct comprising a promoter that is functional in a plant operably linked to (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:4, 6, 8, 10, 12, 14, 16, and 18, or (b) a full-length complement of said polynucleotide of (a)(i); (b) regenerating a transgenic plant from said transformed plant cell; and (c) evaluating said transgenic plant for stalk mechanical strength. The method may further comprise (d) obtaining a progeny plant derived from said transgenic plant; and (e) evaluating said progeny plant for stalk mechanical strength.
[0013]In another embodiment, a method of evaluating stalk mechanical strength in a plant, comprising (a) introducing into a regenerable plant cell a recombinant DNA construct to produce transformed plant cells, said recombinant DNA construct comprising a promoter that is functional in a plant operably linked to (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:4, 6, 8, 10, 12, 14, 16, and 18, or (b) a full-length complement of said polynucleotide of (a)(i); (b) regenerating a transgenic plant from said transformed plant cell; (c) obtaining a progeny plant derived from said transgenic plant; and (d) evaluating said progeny plant for stalk mechanical strength.
[0014]The present invention also includes:
[0015]In one embodiment, a plant comprising in its genome: (a) a first recombinant DNA construct comprising at least one promoter that is functional in a plant operably linked to at least one of a first isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, and 18; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, and 17; and (iii) a full-length complement of the polynucleotide of (a)(i) or (a)(ii); and (b) a second recombinant DNA construct comprising at least one promoter that is functional in a plant operably linked to at least one of a second isolated polynucleotide selected from the group consisting of (iv) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, and 42; (v) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:21, 23, 25, 27, 29, 31, 33, 35, 37, 39, and 41; and (vi) a full-length complement of the polynucleotide of (b)(iv) or (b)(v).
[0016]In another embodiment, a plant comprising in its genome at least one regulatory sequence operably linked to: (a) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, and 18; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, and 17; and (iii) a full-length complement of the polynucleotide of (a)(i) or (a)(ii); and (b) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, and 42; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:21, 23, 25, 27, 29, 31, 33, 35, 37, 39, and 41; and (iii) a full-length complement of the polynucleotide of (b)(i) or (b)(ii), and wherein said plant exhibits increased cell wall cellulose content or enhanced growth rate when compared to a control plant not comprising said at least one regulatory sequence operably linked to said (a) and (b).
[0017]In another embodiment, a plant comprising in its genome a suppression DNA construct comprising a promoter functional in a plant operably linked to (a) all or part of (i) a nucleic acid sequence encoding a polypeptide having an amino acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, and 18, or (ii) a full-length complement of the nucleic acid sequence of (a)(i); or (b) a region derived from all or part of a sense strand or antisense strand of a target gene of interest, or any integer from 51% up to and including 100% sequence identity, said region having a nucleic acid sequence of at least 50% sequence identity, based on the Clustal V method of alignment, when compared to said all or part of a sense strand or antisense strand from which said region is derived, and wherein said target gene of interest encodes a polypeptide selected from the group consisting of Bk2, Bk2L1, Bk2L3, Bk2L4, Bk2L5, Bk2L6, Bk2L7, Bk2L8 and Bk2L9, and wherein said plant exhibits reduced stalk mechanical strength when compared to a control plant not comprising said suppression DNA construct.
[0018]In another embodiment, a plant comprising in its genome a suppression DNA construct comprising a promoter functional in a plant operably linked to (a) all or part of (i) a nucleic acid sequence encoding a polypeptide having an amino acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:6, or (ii) a full-length complement of the nucleic acid sequence of (a)(i); or (b) a region derived from all or part of a sense strand or antisense strand of a target gene of interest, said region having a nucleic acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to said all or part of a sense strand or antisense strand from which said region is derived, and wherein said target gene of interest encodes a Bk2L3 polypeptide, and wherein said plant exhibits reduced plant height and/or reduced organ size when compared to a control plant not comprising said suppression DNA construct.
[0019]In another embodiment, a plant comprising in its genome a suppression DNA construct comprising a promoter functional in a plant operably linked to (a) all or part of (i) a nucleic acid sequence encoding a polypeptide having an amino acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:10, or (ii) a full complement of the nucleic acid sequence of (a)(i); or (b) a region derived from all or part of a sense strand or antisense strand of a target gene of interest, said region having a nucleic acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to said all or part of a sense strand or antisense strand from which said region is derived, and wherein said target gene of interest encodes a Bk2L5 polypeptide, and wherein said plant exhibits male sterility when compared to a control plant not comprising said suppression DNA construct.
BRIEF DESCRIPTION OF THE FIGURES AND SEQUENCE LISTINGS
[0020]The invention can be more fully understood from the following detailed description and the accompanying drawings and Sequence Listing which form a part of this application.
[0021]FIGS. 1A-1F show a Clustal V alignment, using default parameters, of the amino acid sequences of the Bk2 and Bk2-like proteins set forth in SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16 and 18.
[0022]FIG. 2 shows a chart setting forth a comparison of the percent identity (and percent divergence in the lower half triangle), using the Clustal V alignment method, between the nine amino acid sequences shown in FIGS. 1A-1F.
[0023]FIG. 3 shows Solexa MPSS® gene expression analysis of gene Bk2.
[0024]FIG. 4 shows the correlation of expression patterns of the Bk2 gene with members of the CesA gene family.
[0025]FIGS. 5A-5B show the correlation among the expression level of all the different Bk2 and CesA genes from maize as studied from Solexa MPSS®.
[0026]FIG. 6 shows the phylogenetic analysis of the Bk2L proteins from maize, BC1L proteins from rice, and COBL proteins from Arabidopsis (NCBI Accession Nos. are in parenthesis). The numbers along the branches are the bootstrap values obtained from a heuristic search over 5,000 replications. The bootstrap values for only the monophyletic groups that were supported >50% of the time are shown. The branch lengths are proportional to the inferred amino acid differences.
[0027]SEQ ID NO:1 is the 1784 bp nucleotide sequence containing the open reading frame (ORF) (nucleotides 89-1438) of the BRITTLE STALK 2 (Bk2) gene from maize flanked by additional untranslated regions (UTR) 5' (nucleotides 1-88) and 3' (nucleotides 1439-1784) to this ORF region.
[0028]SEQ ID NO:2 is the deduced amino acid sequence of a maize BRITTLE STALK 2 (Bk2) polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:1.
[0029]SEQ ID NO:3 is the 3152 bp nucleotide sequence containing the ORF (nucleotides 586-2586) of the BRITTLE STALK 2-Like1 (Bk2L1) gene from maize flanked by additional UTR regions 5' (nucleotides 1-585) and 3' (nucleotides 2587-3152) to this ORF region.
[0030]SEQ ID NO:4 is the deduced amino acid sequence of a maize BRITTLE STALK 2-Like1 (Bk2L1) polypeptide derived from ORF of the nucleotide sequence set forth in SEQ ID NO:3.
[0031]SEQ ID NO:5 is the 2094 bp nucleotide sequence containing the ORF (nucleotides 281-1624) of the BRITTLE STALK 2-Like3 (Bk2L3) gene from maize flanked by additional UTR regions 5' (nucleotides 1-280) and 3' (nucleotides 1625-2094) to this ORF region.
[0032]SEQ ID NO:6 is the deduced amino acid sequence of a maize BRITTLE STALK 2-Like3 (Bk2L3) polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:5.
[0033]SEQ ID NO:7 is the 2102 bp nucleotide sequence containing the ORF (nucleotides 326-1672) of the BRITTLE STALK 2-Like4 (Bk2L4) gene from maize flanked by additional UTR regions 5' (nucleotides 1-325) and 3' (nucleotides 1673-2102) to this ORF region.
[0034]SEQ ID NO:8 is the deduced amino acid sequence of a maize BRITTLE STALK 2-Like4 (Bk2L4) polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:7.
[0035]SEQ ID NO:9 is the 2422 bp nucleotide sequence containing the ORF (nucleotides 216-2249) of the BRITTLE STALK 2-Like5 (Bk2L5) gene from maize flanked by additional UTR regions 5' (nucleotides 1-215) and 3' (nucleotides 2250-2422) to this ORF region.
[0036]SEQ ID NO:10 is the deduced amino acid sequence of a maize BRITTLE STALK 2-Like5 (Bk2L5) polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:9.
[0037]SEQ ID NO:11 is the 1845 bp nucleotide sequence containing the ORF (nucleotides 184-1563) of the BRITTLE STALK 2-Like6 (Bk2L6) gene from maize flanked by additional UTR regions 5' (nucleotides 1-183) and 3' (nucleotides 1564-1845) to this ORF region.
[0038]SEQ ID NO:12 is the deduced amino acid sequence of a maize BRITTLE STALK 2-Like6 (Bk2L6) polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:11.
[0039]SEQ ID NO:13 is the 1644 bp nucleotide sequence containing the ORF (nucleotides 85-1425) of the BRITTLE STALK 2-Like7 (Bk2L7) gene from maize flanked by additional UTR regions 5' (nucleotides 1-84) and 3' (nucleotides 1426-1644) to this ORF region.
[0040]SEQ ID NO:14 is the deduced amino acid sequence of a maize BRITTLE STALK 2-Like7 (Bk2L7) polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:13.
[0041]SEQ ID NO:15 is the 2108 bp nucleotide sequence containing the ORF (nucleotides 144-2105) of the BRITTLE STALK 2-Like8 (Bk2L8) gene from maize flanked by additional UTR regions 5' (nucleotides 1-143) and 3' (nucleotides 2106-2108) to this ORF region.
[0042]SEQ ID NO:16 is the deduced amino acid sequence of a maize BRITTLE STALK 2-Like8 (Bk2L8) polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:15.
[0043]SEQ ID NO:17 is the 1335 bp nucleotide sequence containing the ORF (nucleotides 1-1332) of the BRITTLE STALK 2-Like9 (Bk2L9) gene from maize flanked by additional UTR regions 5' (nucleotides 0) and 3' (nucleotides 1963-1965) to this ORF region.
[0044]SEQ ID NO:18 is the deduced amino acid sequence of a maize BRITTLE STALK 2-Like9 (Bk2L9) polypeptide derived from the nucleotide sequence set forth in SEQ ID NO:17.
[0045]SEQ ID NO:19 is the 3780 bp nucleotide sequence containing the ORF (nucleotides 201-3428) of the CesA1 gene from maize flanked by additional UTR regions 5' (nucleotides 1-200) and 3' (nucleotides 3429-3780) to this ORF region.
[0046]SEQ ID NO:20 is the deduced amino acid sequence of a maize CesA1 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:19.
[0047]SEQ ID NO:21 is the 3725 bp nucleotide sequence containing the ORF (nucleotides 179-3403) of the CesA2 gene from maize flanked by additional UTR regions 5' (nucleotides 1-178) and 3' (nucleotides 3404-3725) to this ORF region.
[0048]SEQ ID NO:22 is the deduced amino acid sequence of a maize CesA2 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:21.
[0049]SEQ ID NO:23 is the 2830 bp nucleotide sequence containing the ORF (nucleotides 3-2468) of the CesA3 gene from maize flanked by additional UTR regions 5' (nucleotides 1-2) and 3' (nucleotides 2469-2830) to this ORF region.
[0050]SEQ ID NO:24 is the deduced amino acid sequence of a maize CesA3 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:23.
[0051]SEQ ID NO:25 is the 3773 bp nucleotide sequence containing the ORF (nucleotides 338-3571) of the CesA4 gene from maize flanked by additional UTR regions 5' (nucleotides 1-337) and 3' (nucleotides 3572-3773) to this ORF region.
[0052]SEQ ID NO:26 is the deduced amino acid sequence of a maize CesA4 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:25.
[0053]SEQ ID NO:27 is the 3704 bp nucleotide sequence containing the ORF (nucleotides 272-3502) of the CesA5 gene from maize flanked by additional UTR regions 5' (nucleotides 1-271) and 3' (nucleotides 3503-3704) to this ORF region.
[0054]SEQ ID NO:28 is the deduced amino acid sequence of a maize CesA5 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:27.
[0055]SEQ ID NO:29 is the 3568 bp nucleotide sequence containing the ORF (nucleotides 63-3242) of the CesA6 gene from maize flanked by additional UTR regions 5' (nucleotides 1-62) and 3' (nucleotides 3243-3568) to this ORF region.
[0056]SEQ ID NO:30 is the deduced amino acid sequence of a maize CesA6 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:29.
[0057]SEQ ID NO:31 is the 3969 bp nucleotide sequence containing the ORF (nucleotides 144-3404) of the CesA7 gene from maize flanked by additional UTR regions 5' (nucleotides 1-143) and 3' (nucleotides 3405) to this ORF region.
[0058]SEQ ID NO:32 is the deduced amino acid sequence of a maize CesA7 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:31.
[0059]SEQ ID NO:33 is the 3813 bp nucleotide sequence containing the ORF (nucleotides 215-3499) of the CesA8 gene from maize flanked by additional UTR regions 5' (nucleotides 1-214) and 3' (nucleotides 3500-3813) to this ORF region.
[0060]SEQ ID NO:34 is the deduced amino acid sequence of a maize CesA8 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:33.
[0061]SEQ ID NO:35 is the 3799 bp nucleotide sequence containing the ORF (nucleotides 238-3477) of the CesA9 gene from maize flanked by additional UTR regions 5' (nucleotides 1-237) and 3' (nucleotides 3478-3799) to this ORF region.
[0062]SEQ ID NO:36 is the deduced amino acid sequence of a maize CesA9 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:35.
[0063]SEQ ID NO:37 is the 3470 bp nucleotide sequence containing the ORF (nucleotides 29-3265) of the CesA10 gene from maize flanked by additional UTR regions 5' (nucleotides 1-28) and 3' (nucleotides 3266-3470) to this ORF region.
[0064]SEQ ID NO:38 is the deduced amino acid sequence of a maize CesA10 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:37.
[0065]SEQ ID NO:39 is the 3231 bp nucleotide sequence containing the ORF (nucleotides 21-3044) of the CesA11 gene from maize flanked by additional UTR regions 5' (nucleotides 1-20) and 3' (nucleotides 3045-3231) to this ORF region.
[0066]SEQ ID NO:40 is the deduced amino acid sequence of a maize CesA11 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:39.
[0067]SEQ ID NO:41 is the 3028 bp nucleotide sequence containing the ORF (nucleotides 52-2835) of the CesA12 gene from maize flanked by additional UTR regions 5' (nucleotides 1-51) and 3' (nucleotides 2836-3028) to this ORF region.
[0068]SEQ ID NO:42 is the deduced amino acid sequence of a maize CesA12 polypeptide derived from the ORF of the nucleotide sequence set forth in SEQ ID NO:41.
[0069]The Sequence Listing contains the one letter code for nucleotide sequence characters and the three letter codes for amino acids as defined in conformity with the IUPAC-IUBMB standards described in Nucleic Acids Res. 13:3021-3030 (1985) and in the Biochemical J. 219(2):345-373 (1984) which are herein incorporated by reference. The symbols and format used for nucleotide and amino acid sequence data comply with the rules set forth in 37 C.F.R. §1.822. The sequence descriptions and Sequence Listing attached hereto comply with the rules governing nucleotide and/or amino acid sequence disclosures in patent applications as set forth in 37 C.F.R. §1.821-1.825.
DETAILED DESCRIPTION OF THE INVENTION
[0070]All patents, patent applications, and publications cited throughout the application are hereby incorporated by reference in their entirety.
[0071]As used herein and in the appended claims, the singular forms "a", "an", and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, reference to "a plant" includes a plurality of such plants, reference to "a cell" includes one or more cells and equivalents thereof known to those skilled in the art, and so forth.
[0072]In the context of this disclosure, a number of terms shall be utilized.
[0073]"Transgenic" includes any cell, cell line, callus, tissue, plant part or plant, the genome of which has been altered by the presence of a heterologous nucleic acid, such as a recombinant DNA construct, including those transgenics initially so altered as well as those created by sexual crosses or asexual propagation from the initial transgenic. The term "transgenic" as used herein does not encompass the alteration of the genome (chromosomal or extra-chromosomal) by conventional plant breeding methods or by naturally occurring events such as random cross-fertilization, non-recombinant viral infection, non-recombinant bacterial transformation, non-recombinant transposition, or spontaneous mutation.
[0074]"Genome" as it applies to plant cells encompasses not only chromosomal DNA found within the nucleus, but organelle DNA found within subcellular components (e.g., mitochondrial, plastid) of the cell.
[0075]"Plant" includes reference to whole plants, plant organs, plant tissues, seeds and plant cells and progeny of same. Plant cells include, without limitation, cells from seeds, suspension cultures, embryos, meristematic regions, callus tissue, leaves, roots, shoots, gametophytes, sporophytes, pollen, and microspores.
[0076]"Progeny" comprises any subsequent generation of a plant.
[0077]"Transgenic plant" includes reference to a plant which comprises within its genome a heterologous polynucleotide. Preferably, the heterologous polynucleotide is stably integrated within the genome such that the polynucleotide is passed on to successive generations. The heterologous polynucleotide may be integrated into the genome alone or as part of a recombinant DNA construct.
[0078]"Heterologous" with respect to sequence means a sequence that originates from a foreign species, or, if from the same species, is substantially modified from its native form in composition and/or genomic locus by deliberate human intervention.
[0079]"Polynucleotide", "nucleic acid sequence", "nucleotide sequence", or "nucleic acid fragment" are used interchangeably and is a polymer of RNA or DNA that is single or double-stranded, optionally containing synthetic, non-natural or altered nucleotide bases. Nucleotides (usually found in their 5'-monophosphate form) are referred to by their single letter designation as follows: "A" for adenylate or deoxyadenylate (for RNA or DNA, respectively), "C" for cytidylate or deoxycytidylate, "G" for guanylate or deoxyguanylate, "U" for uridylate, "T" for deoxythymidylate, "R" for purines (A or G), "Y" for pyrimidines (C or T), "K" for G or T, "H" for A or C or T, "I" for inosine, and "N" for any nucleotide.
[0080]"Polypeptide", "peptide", "amino acid sequence" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residue is an artificial chemical analogue of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers. The terms "polypeptide", "peptide", "amino acid sequence", and "protein" are also inclusive of modifications including, but not limited to, glycosylation, lipid attachment, sulfation, gamma-carboxylation of glutamic acid residues, hydroxylation and ADP-ribosylation.
[0081]"Messenger RNA (mRNA)" refers to the RNA that is without introns and that can be translated into protein by the cell.
[0082]"cDNA" refers to a DNA that is complementary to and synthesized from a mRNA template using the enzyme reverse transcriptase. The cDNA can be single-stranded or converted into the double-stranded form using the Klenow fragment of DNA polymerase I.
[0083]"Mature" protein refers to a post-translationally processed polypeptide; i.e., one from which any pre- or pro-peptides present in the primary translation product have been removed.
[0084]"Precursor" protein refers to the primary product of translation of mRNA; i.e., with pre- and pro-peptides still present. Pre- and pro-peptides may be and are not limited to intracellular localization signals.
[0085]"Isolated" refers to materials, such as nucleic acid molecules and/or proteins, which are substantially free or otherwise removed from components that normally accompany or interact with the materials in a naturally occurring environment. Isolated polynucleotides may be purified from a host cell in which they naturally occur. Conventional nucleic acid purification methods known to skilled artisans may be used to obtain isolated polynucleotides. The term also embraces recombinant polynucleotides and chemically synthesized polynucleotides.
[0086]"Recombinant" refers to an artificial combination of two otherwise separated segments of sequence, e.g., by chemical synthesis or by the manipulation of isolated segments of nucleic acids by genetic engineering techniques. "Recombinant" also includes reference to a cell or vector, that has been modified by the introduction of a heterologous nucleic acid or a cell derived from a cell so modified, but does not encompass the alteration of the cell or vector by naturally occurring events (e.g., spontaneous mutation, natural transformation/transduction/transposition) such as those occurring without deliberate human intervention.
[0087]"Recombinant DNA construct" refers to a combination of nucleic acid fragments that are not normally found together in nature. Accordingly, a recombinant DNA construct may comprise regulatory sequences and coding sequences that are derived from different sources, or regulatory sequences and coding sequences derived from the same source, but arranged in a manner different than that normally found in nature.
[0088]"Regulatory sequences" refer to nucleotide sequences located upstream (5' non-coding sequences), within, or downstream (3' non-coding sequences) of a coding sequence, and which influence the transcription, RNA processing or stability, or translation of the associated coding sequence. Regulatory sequences may include, but are not limited to, promoters, translation leader sequences, introns, and polyadenylation recognition sequences.
[0089]"Promoter" refers to a nucleic acid fragment capable of controlling transcription of another nucleic acid fragment.
[0090]"Promoter functional in a plant" is a promoter capable of controlling transcription in plant cells whether or not its origin is from a plant cell.
[0091]"Operably linked" refers to the association of nucleic acid fragments in a single fragment so that the function of one is regulated by the other. For example, a promoter is operably linked with a nucleic acid fragment when it is capable of regulating the transcription of that nucleic acid fragment.
[0092]"Expression" refers to the production of a functional product. For example, expression of a nucleic acid fragment may refer to transcription of the nucleic acid fragment (e.g., transcription resulting in mRNA or functional RNA) and/or translation of mRNA into a precursor or mature protein.
[0093]"Phenotype" means the detectable characteristics of a cell or organism.
[0094]"Introduced" in the context of inserting a nucleic acid fragment (e.g., a recombinant DNA construct) into a cell, means "transfection" or "transformation" or "transduction" and includes reference to the incorporation of a nucleic acid fragment into a eukaryotic or prokaryotic cell where the nucleic acid fragment may be incorporated into the genome of the cell (e.g., chromosome, plasmid, plastid or mitochondrial DNA), converted into an autonomous replicon, or transiently expressed (e.g., transfected mRNA).
[0095]A "transformed cell" is any cell into which a nucleic acid fragment (e.g., a recombinant DNA construct) has been introduced.
[0096]"Transformation" as used herein refers to both stable transformation and transient transformation.
[0097]"Stable transformation" refers to the introduction of a nucleic acid fragment into a genome of a host organism resulting in genetically stable inheritance. Once stably transformed, the nucleic acid fragment is stably integrated in the genome of the host organism and any subsequent generation.
[0098]"Transient transformation" refers to the introduction of a nucleic acid fragment into the nucleus, or DNA-containing organelle, of a host organism resulting in gene expression without genetically stable inheritance.
[0099]"Allele" is one of several alternative forms of a gene occupying a given locus on a chromosome. Different alleles of a gene differ in their DNA sequence. When the alleles present at a given locus on a pair of homologous chromosomes in a diploid plant are the same that plant is homozygous at that locus. If the alleles present at a given locus on a pair of homologous chromosomes in a diploid plant differ that plant is heterozygous at that locus. If a transgene is present on one of a pair of homologous chromosomes in a diploid plant that plant is hemizygous at that locus.
[0100]"Contig" refers to a nucleotide sequence that is assembled from two or more constituent nucleotide sequences that share common or overlapping regions of sequence homology. For example, the nucleotide sequences of two or more nucleic acid fragments can be compared and aligned in order to identify common or overlapping sequences. Where common or overlapping sequences exist between two or more nucleic acid fragments, the sequences (and thus their corresponding nucleic acid fragments) can be assembled into a single contiguous nucleotide sequence.
[0101]"Codon degeneracy" refers to divergence in the genetic code permitting variation of the nucleotide sequence without affecting the amino acid sequence of an encoded polypeptide. Accordingly, the instant invention relates to any nucleic acid fragment comprising a nucleotide sequence that encodes all or a substantial portion of the amino acid sequences set forth herein. The skilled artisan is well aware of the "codon-bias" exhibited by a specific host cell in usage of nucleotide codons to specify a given amino acid. Therefore, when synthesizing a nucleic acid fragment for improved expression in a host cell, it is desirable to design the nucleic acid fragment such that its frequency of codon usage approaches the frequency of preferred codon usage of the host cell.
[0102]"Synthetic nucleic acid fragments" can be assembled from oligonucleotide building blocks that are chemically synthesized using procedures known to those skilled in the art. These building blocks are ligated and annealed to form larger nucleic acid fragments which may then be enzymatically assembled to construct the entire desired nucleic acid fragment. "Chemically synthesized", as related to a nucleic acid fragment, means that the component nucleotides were assembled in vitro. Manual chemical synthesis of nucleic acid fragments may be accomplished using well-established procedures, or automated chemical synthesis can be performed using one of a number of commercially available machines. Accordingly, the nucleic acid fragments can be tailored for optimal gene expression based on optimization of the nucleotide sequence to reflect the codon bias of the host cell. The skilled artisan appreciates the likelihood of successful gene expression if codon usage is biased towards those codons favored by the host. Determination of preferred codons can be based on a survey of genes derived from the host cell where sequence information is available.
[0103]The term "amplified" means the construction of multiple copies of a nucleic acid sequence or multiple copies complementary to the nucleic acid sequence using at least one of the nucleic acid sequences as a template. Amplification systems include the polymerase chain reaction (PCR) system, ligase chain reaction (LCR) system, nucleic acid sequence based amplification (NASBA, Cangene, Mississauga, Ontario), Q-Beta Replicase systems, transcription-based amplification system (TAS), and strand displacement amplification (SDA). See, e.g., Diagnostic Molecular Microbiology Principles and Applications, D. H. Persing et al., Ed., American Society for Microbiology, Washington, D.C. (1993). The product of amplification is termed an amplicon.
[0104]The term "chromosomal location" includes reference to a length of a chromosome which may be measured by reference to the linear segment of DNA which it comprises. The chromosomal location can be defined by reference to two unique DNA sequences, i.e., markers.
[0105]The term "marker" includes reference to a locus on a chromosome that serves to identify a unique position on the chromosome. A "polymorphic marker" includes reference to a marker which appears in multiple forms (alleles) such that different forms of the marker, when they are present in a homologous pair, allow transmission of each of the chromosomes in that pair to be followed. A genotype may be defined by use of one or a plurality of markers.
[0106]Sequence alignments and percent identity calculations may be determined using a variety of comparison methods designed to detect homologous sequences including, but not limited to, the Megalign program of the LASARGENE bioinformatics computing suite (DNASTAR Inc., Madison, Wis.). Unless stated otherwise, multiple alignment of the sequences provided herein were performed using the Clustal V method of alignment (Higgins and Sharp (1989) CABIOS. 5:151-153) with the default parameters (GAP PENALTY=10, GAP LENGTH PENALTY=10). Default parameters for pairwise alignments and calculation of percent identity of protein sequences using the Clustal V method are KTUPLE=1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5. For nucleic acids these parameters are KTUPLE=2, GAP PENALTY=5, WINDOW=4 and DIAGONALS SAVED=4. After alignment of the sequences, using the Clustal V program, it is possible to obtain "percent identity" and "divergence" values by viewing the "sequence distances" table on the same program; unless stated otherwise, percent identities and divergences provided and claimed herein were calculated in this manner.
[0107]Unless otherwise stated, "BLAST" sequence identity/similarity values provided herein refer to the value obtained using the BLAST 2.0 suite of programs using default parameters (Altschul et al., Nucleic Acids Res. 25:3389-3402 (1997)). Software for performing BLAST analyses is publicly available, e.g., through the National Center for Biotechnology Information. This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are then extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, a cutoff of 100, M=5, N=-4, and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength (W) of 3, an expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989)).
[0108]As used herein, "any integer from 51% up to and including 100%" means 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%".
[0109]As used herein, "any integer from 61% up to and including 100%" means 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%".
[0110]As used herein, "any integer from 81% up to and including 100%" means 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%".
[0111]As used herein, "80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity, or any other integer in between 80% and 100%" means 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.
[0112]Standard recombinant DNA and molecular cloning techniques used herein are well known in the art and are described more fully in Sambrook, J., Fritsch, E. F. and Maniatis, T. Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, 1989 (hereinafter "Sambrook").
[0113]Turning now to Preferred Embodiments:
[0114]Preferred embodiments include isolated polynucleotides and polypeptides, recombinant DNA constructs, compositions (such as plants or seeds) comprising these recombinant DNA constructs, and methods utilizing these recombinant DNA constructs.
[0115]Preferred Isolated Polynucleotides and Polypeptides
[0116]The present invention includes the following preferred isolated polynucleotides and polypeptides:
[0117]In one preferred embodiment, an isolated polynucleotide comprises (a) a nucleotide sequence encoding a polypeptide associated with stalk mechanical strength, wherein said polypeptide has an amino acid sequence of at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity, or any other integer in between 80% and 100%, based on the Clustal V method of alignment, when compared to SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16 or 18; or (b) a complement of the nucleotide sequence of (a), wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary (i.e., a full-length complement of the nucleotide sequence of (a)). Preferably, the polypeptide is associated with maize stalk mechanical strength, and the amino acid sequence of the polypeptide is compared to SEQ ID NOs:16 or 18.
[0118]In another preferred embodiment, an isolated polynucleotide comprises (a) a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, and 17, or (b) a full-length complement of said nucleic acid sequence of (a).
[0119]In another preferred embodiment, an isolated polypeptide associated with stalk mechanical strength comprises an amino acid sequence of at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity, or any other integer in between 80% and 100%, based on the Clustal V method of alignment, when compared to SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16 or 18.
[0120]Several methods may be used to measure the stalk mechanical strength of plants. Preferably, the mechanical strength may be measured with an electromechanical test system. In the case of maize stalk mechanical strength, in a preferred method, the internodes below the ear may be subjected to a three-point bend test using an Instron, Model 4411 (Instron Corporation, 100 Royall Street, Canton, Mass. 02021), with a span-width of 200 mm between the anchoring points and a speed of 200 mm/minute of the third point attached to a load cell; the load needed to break the internode can be used as a measure of mechanical strength (hereinafter "the three-point bend test"). Internodal breaking strength has been shown to be highly correlated with the amount of cellulose per unit length of the maize stalk (see U.S. Patent Application No. 2004068767 A1, herein incorporated by reference).
[0121]A polypeptide is "associated with stalk mechanical strength" in that the absence of the polypeptide in a plant results in a reduction of stalk mechanical strength of the plant when compared to a control plant that expresses the polypeptide.
[0122]A polypeptide is "associated with maize stalk mechanical strength" in that the absence of the polypeptide in a maize plant results in a reduction of stalk mechanical strength of the maize plant when compared to a control maize plant that expresses the polypeptide.
[0123]It is understood, as those skilled in the art will appreciate, that the invention encompasses more than the specific exemplary sequences. Alterations in a nucleic acid fragment which result in the production of a chemically equivalent amino acid at a given site, but do not affect the functional properties of the encoded polypeptide, are well known in the art. For example, a codon for the amino acid alanine, a hydrophobic amino acid, may be substituted by a codon encoding another less hydrophobic residue, such as glycine, or a more hydrophobic residue, such as valine, leucine, or isoleucine. Similarly, changes which result in substitution of one negatively charged residue for another, such as aspartic acid for glutamic acid, or one positively charged residue for another, such as lysine for arginine, can also be expected to produce a functionally equivalent product. Nucleotide changes which result in alteration of the N-terminal and C-terminal portions of the polypeptide molecule would also not be expected to alter the activity of the polypeptide. Each of the proposed modifications is well within the routine skill in the art, as is determination of retention of biological activity of the encoded products.
[0124]Preferred Recombinant DNA Constructs and Suppression DNA Constructs
[0125]The present invention also includes a recombinant DNA construct comprising at least one polynucleotide operably linked to at least one regulatory sequence (e.g., preferably, a promoter that is functional in said plant), wherein said polynucleotide comprises any isolated polynucleotide of the present invention.
[0126]In one preferred embodiment, a recombinant DNA construct comprises a promoter that is functional in a plant operably linked to (a) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, and 18, or (b) a full-length complement of said polynucleotide of (a).
[0127]In anther preferred embodiment, a recombinant DNA construct comprises a promoter that is functional in a plant operably linked to (a) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, and 17, or (b) a full-length complement of said polynucleotide of (a).
[0128]The present invention also includes a suppression DNA construct.
[0129]In one preferred embodiment, a suppression DNA construct comprises a promoter functional in a plant operably linked to (a) all or part of (i) a nucleic acid sequence encoding a polypeptide having an amino acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, and 18, or (ii) a full-length complement of the nucleic acid sequence of (a)(i); or (b) a region derived from all or part of a sense strand or antisense strand of a target gene of interest, said region having a nucleic acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to said all or part of a sense strand or antisense strand from which said region is derived, and wherein said target gene of interest encodes a polypeptide selected from the group consisting of Bk2, Bk2L1, Bk2L3, Bk2L4, Bk2L5, Bk2L6, Bk2L7, Bk2L8 and Bk2L9.
[0130]"Suppression DNA construct" is a recombinant DNA construct which when transformed or stably integrated into the genome of the plant, results in "silencing" of a target gene in the plant. The target gene may be endogenous or transgenic to the plant. "Silencing," as used herein with respect to the target gene, refers generally to the suppression of levels of mRNA or protein/enzyme expressed by the target gene, and/or the level of the enzyme activity or protein functionality. The term "suppression" includes lower, reduce, decline, decrease, inhibit, eliminate and prevent. "Silencing" or "gene silencing" does not specify mechanism and is inclusive, and not limited to, anti-sense, cosuppression, viral-suppression, hairpin suppression, stem-loop suppression, RNAi-based approaches, and small RNA-based approaches.
[0131]A suppression DNA construct may comprise a region derived from a target gene of interest and may comprise all or part of the nucleic acid sequence of the sense strand (or antisense strand) of the target gene of interest. Depending upon the approach to be utilized, the region may be 100% identical or less than 100% identical (e.g., at least 50% or any integer between 51% and 100% identical) to all or part of the sense strand (or antisense strand) of the gene of interest.
[0132]Suppression DNA constructs are well-known in the art, are readily constructed once the target gene of interest is selected, and include, without limitation, cosuppression constructs, antisense constructs, viral-suppression constructs, hairpin suppression constructs, stem-loop suppression constructs, double-stranded RNA-producing constructs, and more generally, RNAi (RNA interference) constructs and small RNA constructs such as siRNA (short interfering RNA) constructs and miRNA (microRNA) constructs.
[0133]"Antisense inhibition" refers to the production of antisense RNA transcripts capable of suppressing the expression of the target protein.
"Antisense RNA" refers to an RNA transcript that is complementary to all or part of a target primary transcript or mRNA and that blocks the expression of a target isolated nucleic acid fragment (U.S. Pat. No. 5,107,065). The complementarity of an antisense RNA may be with any part of the specific gene transcript, i.e., at the 5' non-coding sequence, 3' non-coding sequence, introns, or the coding sequence.
[0134]"Cosuppression" refers to the production of sense RNA transcripts capable of suppressing the expression of the target protein. "Sense" RNA refers to RNA transcript that includes the mRNA and can be translated into protein within a cell or in vitro. Cosuppression constructs in plants have been previously designed by focusing on overexpression of a nucleic acid sequence having homology to a native mRNA, in the sense orientation, which results in the reduction of all RNA having homology to the overexpressed sequence (see Vaucheret et al. (1998) Plant J. 16:651-659; and Gura (2000) Nature 404:804-808).
[0135]Another variation describes the use of plant viral sequences to direct the suppression of proximal mRNA encoding sequences (PCT Publication WO 98/36083 published on Aug. 20, 1998).
[0136]Recent work has described the use of "hairpin" structures that incorporate all, or part, of an mRNA encoding sequence in a complementary orientation that results in a potential "stem-loop" structure for the expressed RNA (PCT Publication WO 99/53050 published on Oct. 21, 1999). In this case the stem is formed by polynucleotides corresponding to the gene of interest inserted in either sense or anti-sense orientation with respect to the promoter and the loop is formed by some polynucleotides of the gene of interest, which do not have a complement in the construct. This increases the frequency of cosuppression or silencing in the recovered transgenic plants. For review of hairpin suppression see Wesley, S. V. et al. (2003) Methods in Molecular Biology, Plant Functional Genomics: Methods and Protocols 236:273-286.
[0137]A construct where the stem is formed by at least 30 nucleotides from a gene to be suppressed and the loop is formed by a random nucleotide sequence has also effectively been used for suppression (WO 99/61632 published on Dec. 2, 1999).
[0138]The use of poly-T and poly-A sequences to generate the stem in the stem-loop structure has also been described (WO 02/00894 published Jan. 3, 2002).
[0139]Yet another variation includes using synthetic repeats to promote formation of a stem in the stem-loop structure. Transgenic organisms prepared with such recombinant DNA fragments have been shown to have reduced levels of the protein encoded by the nucleotide fragment forming the loop as described in PCT Publication WO 02/00904, published 3 Jan. 2002.
[0140]RNA interference refers to the process of sequence-specific post-transcriptional gene silencing in animals mediated by short interfering RNAs (siRNAs) (Fire et al., Nature 391:806 1998). The corresponding process in plants is commonly referred to as post-transcriptional gene silencing (PTGS) or RNA silencing and is also referred to as quelling in fungi. The process of post-transcriptional gene silencing is thought to be an evolutionarily-conserved cellular defense mechanism used to prevent the expression of foreign genes and is commonly shared by diverse flora and phyla (Fire et al., Trends Genet. 15:358 1999). Such protection from foreign gene expression may have evolved in response to the production of double-stranded RNAs (dsRNAs) derived from viral infection or from the random integration of transposon elements into a host genome via a cellular response that specifically destroys homologous single-stranded RNA of viral genomic RNA. The presence of dsRNA in cells triggers the RNAi response through a mechanism that has yet to be fully characterized.
[0141]The presence of long dsRNAs in cells stimulates the activity of a ribonuclease III enzyme referred to as dicer. Dicer is involved in the processing of the dsRNA into short pieces of dsRNA known as short interfering RNAs (siRNAs) (Berstein et al., Nature 409:363 (2001)). Short interfering RNAs derived from dicer activity are typically about 21 to about 23 nucleotides in length and comprise about 19 base pair duplexes (Elbashir et al., Genes Dev. 15:188 (2001)). Dicer has also been implicated in the excision of 21- and 22-nucleotide small temporal RNAs (stRNAs) from precursor RNA of conserved structure that are implicated in translational control (Hutvagner et al., Science 293:834 (2001)). The RNAi response also features an endonuclease complex, commonly referred to as an RNA-induced silencing complex (RISC), which mediates cleavage of single-stranded RNA having sequence complementarity to the antisense strand of the siRNA duplex. Cleavage of the target RNA takes place in the middle of the region complementary to the antisense strand of the siRNA duplex (Elbashir et al., Genes Dev. 15:188 (2001)). In addition, RNA interference can also involve small RNA (e.g., miRNA) mediated gene silencing, presumably through cellular mechanisms that regulate chromatin structure and thereby prevent transcription of target gene sequences (see, e.g., Allshire, Science 297:1818-1819 (2002); Volpe et al., Science 297:1833-1837 (2002); Jenuwein, Science 297:2215-2218 (2002); and Hall et al., Science 297:2232-2237 (2002)). As such, miRNA molecules of the invention can be used to mediate gene silencing via interaction with RNA transcripts or alternately by interaction with particular gene sequences, wherein such interaction results in gene silencing either at the transcriptional or post-transcriptional level.
[0142]RNAi has been studied in a variety of systems. Fire et al. (Nature 391:806 (1998)) were the first to observe RNAi in C. elegans. Wianny and Goetz (Nature Cell Biol. 2:70 (1999)) describe RNAi mediated by dsRNA in mouse embryos. Hammond et al. (Nature 404:293 (2000)) describe RNAi in Drosophila cells transfected with dsRNA. Elbashir et al., (Nature 411:494 (2001)) describe RNAi induced by introduction of duplexes of synthetic 21-nucleotide RNAs in cultured mammalian cells including human embryonic kidney and HeLa cells.
[0143]Small RNAs play an important role in controlling gene expression. Regulation of many developmental processes, including flowering, is controlled by small RNAs. It is now possible to engineer changes in gene expression of plant genes by using transgenic constructs which produce small RNAs in the plant.
[0144]Small RNAs appear to function by base-pairing to complementary RNA or DNA target sequences. When bound to RNA, small RNAs trigger either RNA cleavage or translational inhibition of the target sequence. When bound to DNA target sequences, it is thought that small RNAs can mediate DNA methylation of the target sequence. The consequence of these events, regardless of the specific mechanism, is that gene expression is inhibited.
[0145]It is thought that sequence complementarity between small RNAs and their RNA targets helps to determine which mechanism, RNA cleavage or translational inhibition, is employed. It is believed that siRNAs, which are perfectly complementary with their targets, work by RNA cleavage. Some miRNAs have perfect or near-perfect complementarity with their targets, and RNA cleavage has been demonstrated for at least a few of these miRNAs. Other miRNAs have several mismatches with their targets, and apparently inhibit their targets at the translational level. Again, without being held to a particular theory on the mechanism of action, a general rule is emerging that perfect or near-perfect complementarity causes RNA cleavage, whereas translational inhibition is favored when the miRNA/target duplex contains many mismatches. The apparent exception to this is microRNA 172 (miR172) in plants. One of the targets of miR172 is APETALA2 (AP2), and although miR172 shares near-perfect complementarity with AP2 it appears to cause translational inhibition of AP2 rather than RNA cleavage.
[0146]MicroRNAs (miRNAs) are noncoding RNAs of about 19 to about 24 nucleotides (nt) in length that have been identified in both animals and plants (Lagos-Quintana et al., Science 294:853-858 (2001), Lagos-Quintana et al., Curr. Biol. 12:735-739 (2002); Lau et al., Science 294:858-862 (2001); Lee and Ambros, Science 294:862-864 (2001); Llave et al., Plant Cell 14:1605-1619 (2002); Mourelatos et al., Genes. Dev. 16:720-728 (2002); Park et al., Curr. Biol. 12:1484-1495 (2002); Reinhart et al., Genes. Dev. 16:1616-1626 (2002)). They are processed from longer precursor transcripts that range in size from approximately 70 to 200 nt, and these precursor transcripts have the ability to form stable hairpin structures. In animals, the enzyme involved in processing miRNA precursors is called Dicer, an RNAse III-like protein (Grishok et al., Cell 106:23-34 2001; Hutvagner et al., Science 293:834-838 (2001); Ketting et al., Genes. Dev. 15:2654-2659 (2001)). Plants also have a Dicer-like enzyme, DCL1 (previously named CARPEL FACTORY/SHORT INTEGUMENTS1/SUSPENSOR1), and recent evidence indicates that it, like Dicer, is involved in processing the hairpin precursors to generate mature miRNAs (Park et al., Curr. Biol. 12:1484-1495 (2002); Reinhart et al., Genes. Dev. 16:1616-1626 (2002)). Furthermore, it is becoming clear from recent work that at least some miRNA hairpin precursors originate as longer polyadenylated transcripts, and several different miRNAs and associated hairpins can be present in a single transcript (Lagos-Quintana et al., Science 294:853-858 (2001); Lee et al., EMBO J. 21:4663-4670 2002). Recent work has also examined the selection of the miRNA strand from the dsRNA product arising from processing of the hairpin by DICER (Schwartz, et al. Cell 115:199-208 (2003)). It appears that the stability (i.e. G:C vs. A:U content, and/or mismatches) of the two ends of the processed dsRNA affects the strand selection, with the low stability end being easier to unwind by a helicase activity. The 5' end strand at the low stability end is incorporated into the RISC complex, while the other strand is degraded.
[0147]MicroRNAs appear to regulate target genes by binding to complementary sequences located in the transcripts produced by these genes. In the case of lin-4 and let-7, the target sites are located in the 3' UTRs of the target mRNAs (Lee et al., Cell 75:843-854 (1993); Wightman et al., Cell 75:855-862 (1993); Reinhart et al., Nature 403:901-906 (2000); Slack et al., Mol. Cell. 5:659-669 (2000)), and there are several mismatches between the lin-4 and let-7 miRNAs and their target sites. Binding of the lin-4 or let-7 miRNA appears to cause downregulation of steady-state levels of the protein encoded by the target mRNA without affecting the transcript itself (Olsen and Ambros, Dev. Biol. 216:671-680 (1999)). On the other hand, recent evidence suggests that miRNAs can in some cases cause specific RNA cleavage of the target transcript within the target site, and this cleavage step appears to require 100% complementarity between the miRNA and the target transcript (Hutvagner and Zamore, Science 297:2056-2060 (2002); Llave et al., Plant Cell 14:1605-1619 (2002)). It seems likely that miRNAs can enter at least two pathways of target gene regulation: Protein downregulation when target complementarity is <100%, and RNA cleavage when target complementarity is 100%. MicroRNAs entering the RNA cleavage pathway are analogous to the 21-25 nt short interfering RNAs (siRNAs) generated during RNA interference (RNAi) in animals and posttranscriptional gene silencing (PTGS) in plants (Hamilton and Baulcombe (1999); Hammond et al., (2000); Zamore et al., (2000); Elbashir et al., (2001)), and likely are incorporated into an RNA-induced silencing complex (RISC) that is similar or identical to that seen for RNAi.
[0148]Identifying the targets of miRNAs with bioinformatics has not been successful in animals, and this is probably due to the fact that animal miRNAs have a low degree of complementarity with their targets. On the other hand, bioinformatic approaches have been successfully used to predict targets for plant miRNAs (Llave et al., Plant Cell 14:1605-1619 (2002); Park et al., Curr. Biol. 12:1484-1495 (2002); Rhoades et al., Cell 110:513-520 (2002)), and thus it appears that plant miRNAs have higher overall complementarity with their putative targets than do animal miRNAs. Most of these predicted target transcripts of plant miRNAs encode members of transcription factor families implicated in plant developmental patterning or cell differentiation.
[0149]Preferred Regulatory Elements of Recombinant DNA Constructs and Suppression DNA Constructs
[0150]A number of promoters can be used in recombinant DNA constructs and suppression DNA constructs of the present invention. The promoters can be selected based on the desired outcome, and may include constitutive, tissue-specific, inducible, or other promoters for expression in the host organism.
[0151]High level, constitutive expression of the candidate gene under control of the 35S promoter may have pleiotropic affects. However, tissue specific and/or stress-specific expression may eliminate undesirable affects but retain the ability to enhance drought tolerance. This affect has been observed in Arabidopsis (Kasuga et al., Nature Biotechnol. 17:287-91 (1999)). As such, candidate gene efficacy may be tested when driven by different promoters.
[0152]Suitable constitutive promoters for use in a plant host cell include, for example, the core promoter of the Rsyn7 promoter and other constitutive promoters disclosed in WO 99/43838 and U.S. Pat. No. 6,072,050; the core CaMV 35S promoter (Odell et al., Nature 313:810-812 (1985)); rice actin (McElroy et al., Plant Cell 2:163-171 (1990)); ubiquitin (Christensen et al., Plant Mol. Biol. 12:619-632 (1989) and Christensen et al., Plant Mol. Biol. 18:675-689 (1992)); pEMU (Last et al., Theor. Appl. Genet. 81:581-588 (1991)); MAS (Velten et al., EMBO J. 3:2723-2730 (1984)); ALS promoter (U.S. Pat. No. 5,659,026), and the like. Other constitutive promoters include, for example, those discussed in U.S. Pat. Nos. 5,608,149; 5,608,144; 5,604,121; 5,569,597; 5,466,785; 5,399,680; 5,268,463; 5,608,142; and 6,177,611.
[0153]In choosing a promoter to use in the methods of the invention, it may be desirable to use a tissue-specific or developmentally regulated promoter. A tissue-specific or developmentally regulated promoter is a DNA sequence which regulates the expression of a DNA sequence selectively in the cells/tissues of a plant critical to tassel development, seed set, or both, and limits the expression of such a DNA sequence to the period of tassel development or seed maturation in the plant. Any identifiable promoter may be used in the methods of the present invention which causes the desired temporal and spatial expression.
[0154]A preferred stalk-specific promoter is the alfalfa stalk-specific S2A gene (Abrahams et al., Plant Mol. Biol. 27:513-528 (1995))
[0155]Promoters which are seed or embryo specific and may be useful in the invention include soybean Kunitz trysin inhibitor (Kti3, Jofuku and Goldberg, Plant Cell 1:1079-1093 (1989)), patatin (potato tubers) (Rocha-Sosa et al., EMBO J. 8:23-29 (1989)), convicilin, vicilin, and legumin (pea cotyledons) (Rerie, W. G., et al. Mol. Gen. Genet. 259:149-157 (1991); Newbigin, E. J., et al., Planta 180:461-470 (1990); Higgins, T. J. V., et al., Plant. Mol. Biol. 11:683-695 (1988)), zein (maize endosperm) (Schemthaner, J. P., et al., EMBO J. 7:1249-1255 (1988)), phaseolin (bean cotyledon) (Segupta-Gopalan, C., et al., Proc. Natl. Acad. Sci. U.S.A. 82:3320-3324 (1985)), phytohemagglutinin (bean cotyledon) (Voelker, T. et al., EMBO J. 6:3571-3577 (1987)), B-conglycinin and glycinin (soybean cotyledon) (Chen, Z-L, et al., EMBO J. 7:297-302 (1988)), glutelin (rice endosperm), hordein (barley endosperm) (Marris, C., et al., Plant Mol. Biol. 10:359-366 (1988)), glutenin and gliadin (wheat endosperm) (Colot, V., et al., EMBO J. 6:3559-3564 (1987)), and sporamin (sweet potato tuberous root) (Hattori, T., et al., Plant Mol. Biol. 14:595-604 (1990)). Promoters of seed-specific genes operably linked to heterologous coding regions in chimeric gene constructions maintain their temporal and spatial expression pattern in transgenic plants. Such examples include Arabidopsis thaliana 2S seed storage protein gene promoter to express enkephalin peptides in Arabidopsis and Brassica napus seeds (Vanderkerckhove et al., Bio/Technology 7:L929-932 (1989)), bean lectin and bean beta-phaseolin promoters to express luciferase (Riggs et al., Plant Sci. 63:47-57 (1989)), and wheat glutenin promoters to express chloramphenicol acetyl transferase (Colot et al., EMBO J. 6:3559-3564 (1987)).
[0156]Inducible promoters selectively express an operably linked DNA sequence in response to the presence of an endogenous or exogenous stimulus, for example by chemical compounds (chemical inducers) or in response to environmental, hormonal, chemical, and/or developmental signals. Inducible or regulated promoters include, for example, promoters regulated by light, heat, stress, flooding or drought, phytohormones, wounding, or chemicals such as ethanol, jasmonate, salicylic acid, or safeners.
[0157]Promoters which are timed to stress include the following: 1) the RD29A promoter (Kasuga et al., Nature Biotechnol. 17:287-291 (1991)); 2) barley promoter, B22E; expression of B22E is specific to the pedicel in developing maize kernels ("Primary Structure of a Novel Barley Gene Differentially Expressed in Immature Aleurone Layers". Klemsdae, S. S. et al., Mol. Gen. Genet. 228(1/2):9-16 (1991)); and 3) maize promoter, Zag2 ("Identification and molecular characterization of ZAG1, the maize homolog of the Arabidopsis floral homeotic gene AGAMOUS", Schmidt, R. J. et al., Plant Cell 5(7):729-737 (1993)). Zag2 transcripts can be detected 5 days prior to pollination to 7 to 8 DAP, and directs expression in the carpel of developing female inflorescences and Ciml which is specific to the nucleus of developing maize kernels. Ciml transcript is detected 4 to 5 days before pollination to 6 to 8 DAP. Other useful promoters include any promoter which can be derived from a gene whose expression is maternally associated with developing female florets.
[0158]Promoters may be derived in their entirety from a native gene, or be composed of different elements derived from different promoters found in nature, or even comprise synthetic DNA segments. It is understood by those skilled in the art that different promoters may direct the expression of a gene in different tissues or cell types, or at different stages of development, or in response to different environmental conditions. It is further recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, DNA fragments of some variation may have identical promoter activity. Promoters that cause a gene to be expressed in most cell types at most times are commonly referred to as "constitutive promoters". New promoters of various types useful in plant cells are constantly being discovered; numerous examples may be found in the compilation by Okamuro, J. K., and Goldberg, R. B., Biochemistry of Plants 15:1-82 (1989).
[0159]Particularly preferred promoters may include: alfalfa stalk-specific S2A gene promoter, RIP2, mLIP15, ZmCOR1, Rab17, CaMV 35S, RD29A, SAM synthetase, ubiquitin, CaMV 19S, nos, Adh, sucrose synthase, R-allele, or root cell promoter. Other preferred promoters include any of the CesA10, CesA11, and CesA12 promoters disclosed in United States Patent Publication 2005/0086712A1, which is hereby incorporated by reference in its entirety.
[0160]Recombinant DNA constructs and suppression DNA constructs of the present invention may also include other regulatory sequences, including but not limited to, translation leader sequences, introns, and polyadenylation recognition sequences. In another preferred embodiment of the present invention, a recombinant DNA construct of the present invention further comprises an enhancer or silencer.
[0161]An intron sequence can be added to the 5' untranslated region or the coding sequence of the partial coding sequence to increase the amount of the mature message that accumulates in the cytosol. Inclusion of a spliceable intron in the transcription unit in both plant and animal expression constructs has been shown to increase gene expression at both the mRNA and protein levels up to 1000-fold. Buchman and Berg, Mol. Cell. Biol. 8:4395-4405 (1988); Callis et al., Genes Dev. 1:1183-1200 (1987). Such intron enhancement of gene expression is typically greatest when placed near the 5' end of the transcription unit. Use of maize introns Adh1-S intron 1, 2, and 6, the Bronze-1 intron are known in the art. See generally, The Maize Handbook, Chapter 116, Freeling and Walbot, Eds., Springer, New York (1994).
[0162]If polypeptide expression is desired, it is generally desirable to include a polyadenylation region at the 3'-end of a polynucleotide coding region. The polyadenylation region can be derived from the natural gene, from a variety of other plant genes, or from T-DNA. The 3' end sequence to be added can be derived from, for example, the nopaline synthase or octopine synthase genes, or alternatively from another plant gene, or less preferably from any other eukaryotic gene.
[0163]A translation leader sequence is a DNA sequence located between the promoter sequence of a gene and the coding sequence. The translation leader sequence is present in the fully processed mRNA upstream of the translation start sequence. The translation leader sequence may affect processing of the primary transcript to mRNA, mRNA stability or translation efficiency. Examples of translation leader sequences have been described (Turner, R. and Foster, G. D., Molecular Biotechnology 3:225 (1995)).
[0164]Any plant can be selected for the identification of regulatory sequences and genes to be used in creating recombinant DNA constructs and suppression DNA constructs of the present invention. Examples of suitable plant targets for the isolation of genes and regulatory sequences would include but are not limited to alfalfa, apple, apricot, Arabidopsis, artichoke, arugula, asparagus, avocado, banana, barley, beans, beet, blackberry, blueberry, broccoli, brussels sprouts, cabbage, canola, cantaloupe, carrot, cassaya, castorbean, cauliflower, celery, cherry, chicory, cilantro, citrus, clementines, clover, coconut, coffee, corn, cotton, cranberry, cucumber, Douglas fir, eggplant, endive, escarole, eucalyptus, fennel, figs, garlic, gourd, grape, grapefruit, honey dew, jicama, kiwifruit, lettuce, leeks, lemon, lime, Loblolly pine, linseed, mango, melon, mushroom, nectarine, nut, oat, oil palm, oil seed rape, okra, olive, onion, orange, an ornamental plant, palm, papaya, parsley, parsnip, pea, peach, peanut, pear, pepper, persimmon, pine, pineapple, plantain, plum, pomegranate, poplar, potato, pumpkin, quince, radiata pine, radiscchio, radish, rapeseed, raspberry, rice, rye, sorghum, Southern pine, soybean, spinach, squash, strawberry, sugarbeet, sugarcane, sunflower, sweet potato, sweetgum, tangerine, tea, tobacco, tomato, triticale, turf, turnip, a vine, watermelon, wheat, yams, and zucchini. Particularly preferred plants for the identification of regulatory sequences are Arabidopsis, corn, wheat, soybean, and cotton.
[0165]In another preferred embodiment of the present invention, a recombinant DNA construct of the present invention further comprises an enhancer.
[0166]Preferred Compositions
[0167]A preferred composition of the present invention is a plant comprising in its genome any of the recombinant DNA constructs of the present invention (such as those preferred constructs discussed above).
[0168]Another preferred composition is a plant whose genome comprises a disruption (e.g., an insertion, such as a transposable element, or sequence mutation) of at least one gene (which may be heterologous or endogenous to the genome) selected from the group consisting of Bk2, Bk2L1, Bk2L3, Bk2L4, Bk2L5, Bk2L6, Bk2L7, Bk2L8 and Bk2L9.
[0169]Still another preferred composition is a plant whose genome comprises other recombinant DNA constructs as discussed below (e.g., constructs involving nucleic acid sequences and amino acid sequences relating to SEQ ID NOs:20-42).
[0170]Preferred compositions also include any progeny of the plant, and any seed obtained from the plant or its progeny. Progeny includes subsequent generations obtained by self-pollination or out-crossing of a plant. Progeny also includes hybrids and inbreds.
[0171]Preferably, in hybrid seed propagated crops, mature transgenic plants can be self-crossed to produce a homozygous inbred plant. The inbred plant produces seed containing the newly introduced recombinant DNA construct. These seeds can be grown to produce plants that would contain the recombinant DNA construct in its genome and exhibit the associated phenotype(s) as described herein, or used in a breeding program to produce hybrid seed, which can be grown to produce plants that would contain the recombinant DNA construct and exhibit the associated phenotype(s) as described herein. Preferably, the seeds are maize.
[0172]Preferably, the plant is a monocotyledonous or dicotyledonous plant, more preferably, a maize or soybean plant, even more preferably a maize plant, such as a maize hybrid plant or a maize inbred plant. The plant may also be sunflower, sorghum, canola, wheat, alfalfa, cotton, rice, barley or millet.
[0173]Preferably, any recombinant DNA construct is stably integrated into the genome of the plant.
[0174]Particularly Preferred Embodiments Include:
[0175]1. A plant (preferably maize) comprising in its genome a recombinant DNA construct comprising at least one regulatory element operably linked to (a) a nucleotide sequence encoding a polypeptide associated with stalk mechanical strength, wherein said polypeptide has an amino acid sequence of at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity, or any other integer in between 80% and 100%, based on the Clustal V method of alignment, when compared to SEQ ID NOs:16 or 18; or (b) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary (i.e., a full length complement of the nucleotide sequence of (a)). Preferably, the at least one regulatory element is a promoter that is functional in a plant.
[0176]2. A plant (preferably maize) comprising in its genome (a) a first recombinant DNA construct comprising at least one promoter that is functional in a plant operably linked to at least one of a first isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, and 18; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, and 17; and (iii) a full-length complement of the polynucleotide of (a)(i) or (a)(ii); and (b) a second recombinant DNA construct comprising at least one promoter that is functional in a plant operably linked to at least one of a second isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, and 42; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:21, 23, 25, 27, 29, 31, 33, 35, 37, 39, and 41; and (iii) a full-length complement of the polynucleotide of (b)(i) or (b)(ii). Preferably, the plant exhibits increased cell wall cellulose content and/or enhanced growth rate when compared to a control plant not comprising said first recombinant DNA construct and said second recombinant DNA construct.
[0177]3. A plant (preferably maize) comprising in its genome at least one regulatory sequence operably linked to (a) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, and 18; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, and 17; and (iii) a full-length complement of the polynucleotide of (a)(i) or (a)(ii); and (b) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, and 42; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:21, 23, 25, 27, 29, 31, 33, 35, 37, 39, and 41; and (iii) a full-length complement of the polynucleotide of (b)(i) or (b)(ii). Preferably, the plant exhibits increased cell wall cellulose content and/or enhanced growth rate when compared to a control plant not comprising said at least one regulatory sequence operably linked to said (a) and (b).
[0178]4. A plant (preferably maize) comprising in its genome at least one regulatory sequence operably linked to (a) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:2; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:1; and (iii) a full-length complement of the polynucleotide of (a)(i) or (a)(ii); and (b) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:38, 40, and 42; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:37, 39, and 41; and (iii) a full-length complement of the polynucleotide of (b)(i) or (b)(ii). Preferably, the plant exhibits increased cell wall cellulose content when compared to a control plant not comprising said at least one regulatory sequence operably linked to said (a) and (b).
[0179]5. A plant (preferably maize) comprising in its genome at least one regulatory sequence operably linked to (a) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:6; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:5; and (iii) a full-length complement of the polynucleotide of (a)(i) or (a)(ii); and (b) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:20, 32, and 34; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:19, 31, and 33; and (iii) a full-length complement of the polynucleotide of (b)(i) or (b)(ii). Preferably, the plant exhibits enhanced growth rate when compared to a control plant not comprising said at least one regulatory sequence operably linked to said (a) and (b).
[0180]6. A plant (preferably maize) comprising in its genome at least one regulatory sequence operably linked to at least two isolated polynucleotides selected from the group consisting of (a) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, and 18; (b) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, and 17; and (c) a full-length complement of the polynucleotide of (a) or (b).
[0181]7. A plant (preferably maize) comprising in its genome a suppression DNA construct comprising a promoter functional in a plant operably linked to (a) all or part of (i) a nucleic acid sequence encoding a polypeptide having an amino acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, and 18, or (ii) a full complement of the nucleic acid sequence of (a)(i); or (b) a region derived from all or part of a sense strand or antisense strand of a target gene of interest, said region having a nucleic acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity sequence identity, based on the Clustal V method of alignment, when compared to said all or part of a sense strand or antisense strand from which said region is derived, and wherein said target gene of interest encodes a polypeptide selected from the group consisting of BK2, Bk2L1, Bk2L3, Bk2L4, Bk2L5, Bk2L6, Bk2L7, Bk2L8 and Bk2L9. Preferably, the plant exhibits reduced stalk mechanical strength when compared to a control plant not comprising said suppression DNA construct. Preferably, the suppression DNA construct comprises a cosuppression construct, antisense construct, viral-suppression construct, hairpin suppression construct, stem-loop suppression construct, double-stranded RNA-producing construct, RNAi construct, or small RNA construct (e.g., an siRNA construct or an miRNA construct).
[0182]8. A plant (preferably maize) whose genome comprises a disruption of at least one gene encoding a polypeptide selected from the group consisting of BK2, Bk2L1, Bk2L3, Bk2L4, Bk2L5, Bk2L6, Bk2L7, Bk2L8 and Bk2L9. Preferably, the disruption results in said plant exhibiting reduced stalk mechanical strength when compared to a control plant not comprising said disruption. Preferably, the disruption comprises an insertion, such as a transposable element or sequence mutation.
[0183]9. A plant (preferably maize) comprising in its genome a suppression DNA construct comprising a promoter functional in a plant operably linked to (a) all or part of (i) a nucleic acid sequence encoding a polypeptide having an amino acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:6, or (ii) a full complement of the nucleic acid sequence of (a)(i); or (b) a region derived from all or part of a sense strand or antisense strand of a target gene of interest, said region having a nucleic acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to said all or part of a sense strand or antisense strand from which said region is derived, and wherein said target gene of interest encodes a Bk2L3 polypeptide. Preferably, the plant exhibits reduced plant height and/or reduced organ size when compared to a control plant not comprising said suppression DNA construct. Preferably, the suppression DNA construct comprises a cosuppression construct, antisense construct, viral-suppression construct, hairpin suppression construct, stem-loop suppression construct, double-stranded RNA-producing construct, RNAi construct, or small RNA construct (e.g., an siRNA construct or an miRNA construct).
[0184]10. A plant (preferably maize) whose genome comprises a disruption of at least one gene encoding a Bk2L3 polypeptide. Preferably, said disruption results in said plant exhibiting reduced plant height and/or reduced organ size when compared to a control plant not comprising said disruption. Preferably, the disruption comprises an insertion, such as a transposable element or sequence mutation.
[0185]11. A plant (preferably maize) comprising in its genome a suppression DNA construct comprising a promoter functional in a plant operably linked to (a) all or part of (i) a nucleic acid sequence encoding a polypeptide having an amino acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:10, or (ii) a full complement of the nucleic acid sequence of (a)(i); or (b) a region derived from all or part of a sense strand or antisense strand of a target gene of interest, said region having a nucleic acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to said all or part of a sense strand or antisense strand from which said region is derived, and wherein said target gene of interest encodes a Bk2L5 polypeptide. Preferably, the plant exhibits male sterility when compared to a control plant not comprising said suppression DNA construct. Preferably, the suppression DNA construct comprises a cosuppression construct, antisense construct, viral-suppression construct, hairpin suppression construct, stem-loop suppression construct, double-stranded RNA-producing construct, RNAi construct, or small RNA construct (e.g., an siRNA construct or an miRNA construct).
[0186]12. A plant (preferably maize) whose genome comprises a disruption of at least one gene encoding a BKL5 polypeptide. Preferably, the disruption results in said plant exhibiting male sterility when compared to a control plant not comprising said disruption. Preferably, the disruption comprises an insertion, such as a transposable element or sequence mutation.
[0187]13. Any progeny of the above plants, any seeds of the above plants, any seeds of progeny of the above plants, and cells from any of the above plants and progeny.
[0188]One of ordinary skill in the art would readily recognize a suitable control or reference plant for use in comparing or measuring relative to a plant comprising within its genome a recombinant DNA construct (or suppression DNA construct). For example, by way of non-limiting illustrations: [0189]Progeny of a transformed plant which is hemizygous with respect to a recombinant DNA construct (or suppression DNA construct), such that the progeny are segregating into plants either comprising or not comprising the recombinant DNA construct (or suppression DNA construct): progeny comprising the recombinant DNA construct (or suppression DNA construct) would be typically measured relative to the progeny not comprising the recombinant DNA construct (or suppression DNA construct). [0190]Introgression of a recombinant DNA construct (or suppression DNA construct) into an inbred line, such as in corn, or into a variety, such as in soybean: the introgressed line would typically be measured relative to the parent inbred or variety line. [0191]Two hybrid lines, where the first hybrid line is produced from two parent inbred lines, and the second hybrid line is produced from the same two parent inbred lines except that one of the parent inbred lines contains a recombinant DNA construct (or suppression DNA construct): the second hybrid line would typically be measured relative to the first hybrid line. [0192]A plant comprising a recombinant DNA construct (or suppression DNA construct) in its genome (or a plant comprising a disruption of a gene in its genome): the plant may be measured relative to a control plant not comprising the recombinant DNA construct (or suppression DNA construct) in its genome (or to a control plant not comprising the disruption) but otherwise having a comparable genetic background to the plant (e.g., sharing at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity of nuclear genetic material compared to the plant comprising the recombinant DNA construct or suppression DNA construct or disruption). There are many laboratory-based techniques available for the analysis, comparison and characterization of plant genetic backgrounds; among these are Isozyme Electrophoresis, Restriction Fragment Length Polymorphisms (RFLPs), Randomly Amplified Polymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), and Simple Sequence Repeats (SSRs) which are also referred to as Microsatellites.
[0193]The introduction of recombinant DNA constructs of the present invention into plants may be carried out by any suitable technique, including but not limited to direct DNA uptake, chemical treatment, electroporation, microinjection, cell fusion, infection, vector mediated DNA transfer, bombardment, or Agrobacterium mediated transformation. Where multiple or stacked recombinant DNA constructs or isolated polynucleotides are desired to be integrated into the genome (e.g., to effect co-expression of two or more isolated polynucleotides), the individual isolated polynucleotides may be introduced into parent lines and crossed through traditional breeding techniques to provide the desired combination or stack in subsequent progeny plants.
[0194]Preferred techniques are set forth below in Example 3 for transformation of maize plant cells and in Example 8 for transformation of soybean plant cells.
[0195]Other preferred methods for transforming dicots, primarily by use of Agrobacterium tumefaciens, and obtaining transgenic plants include those published for cotton (U.S. Pat. No. 5,004,863, U.S. Pat. No. 5,159,135, U.S. Pat. No. 5,518,908); soybean (U.S. Pat. No. 5,569,834, U.S. Pat. No. 5,416,011, McCabe et. al., Bio/Technology 6:923 (1988), Christou et al., Plant Physiol. 87:671 674 (1988)); Brassica (U.S. Pat. No. 5,463,174); peanut (Cheng et al., Plant Cell Rep. 15:653 657 (1996), McKently et al., Plant Cell Rep. 14:699 703 (1995)); papaya; and pea (Grant et al., Plant Cell Rep. 15:254 258, (1995)).
[0196]Transformation of monocotyledons using electroporation, particle bombardment, and Agrobacterium have also been reported and are included as preferred methods, for example, transformation and plant regeneration as achieved in asparagus (Bytebier et al., Proc. Natl. Acad. Sci. (USA) 84:5354, (1987)); barley (Wan and Lemaux, Plant Physiol 104:37 (1994)); Zea mays (Rhodes et al., Science 240:204 (1988), Gordon-Kamm et al., Plant Cell 2:603 618 (1990), Fromm et al., Bio/Technology 8:833 (1990), Koziel et al., Bio/Technology 11: 194, (1993), Armstrong et al., Crop Science 35:550 557 (1995)); oat (Somers et al., Bio/Technology 10: 15 89 (1992)); orchard grass (Horn et al., Plant Cell Rep. 7:469 (1988)); rice (Toriyama et al., TheorAppl. Genet. 205:34, (1986); Part et al., Plant Mol. Biol. 32:1135 1148, (1996); Abedinia et al., Aust. J. Plant Physiol. 24:133 141 (1997); Zhang and Wu, Theor. Appl. Genet. 76:835 (1988); Zhang et al. Plant Cell Rep. 7:379, (1988); Battraw and Hall, Plant Sci. 86:191 202 (1992); Christou et al., Bio/Technology 9:957 (1991)); rye (De la Pena et al., Nature 325:274 (1987)); sugarcane (Bower and Birch, Plant J. 2:409 (1992)); tall fescue (Wang et al., Bio/Technology 10:691 (1992)), and wheat (Vasil et al., Bio/Technology 10:667 (1992); U.S. Pat. No. 5,631,152).
[0197]There are a variety of methods for the regeneration of plants from plant tissue. The particular method of regeneration will depend on the starting plant tissue and the particular plant species to be regenerated.
[0198]The regeneration, development, and cultivation of plants from single plant protoplast transformants or from various transformed explants is well known in the art (Weissbach and Weissbach, In: Methods for Plant Molecular Biology, (Eds.), Academic Press, Inc. San Diego, Calif., (1988)). This regeneration and growth process typically includes the steps of selection of transformed cells, culturing those individualized cells through the usual stages of embryonic development through the rooted plantlet stage. Transgenic embryos and seeds are similarly regenerated. The resulting transgenic rooted shoots are thereafter planted in an appropriate plant growth medium such as soil.
[0199]The development or regeneration of plants containing the foreign, exogenous isolated nucleic acid fragment that encodes a protein of interest is well known in the art. Preferably, the regenerated plants are self-pollinated to provide homozygous transgenic plants. Otherwise, pollen obtained from the regenerated plants is crossed to seed-grown plants of agronomically important lines. Conversely, pollen from plants of these important lines is used to pollinate regenerated plants. A transgenic plant of the present invention containing a desired polypeptide is cultivated using methods well known to one skilled in the art.
Assays to detect proteins may be performed by SDS-polyacrylamide gel electrophoresis or immunological assays. Assays to detect levels of substrates or products of enzymes may be performed using gas chromatography or liquid chromatography for separation and UV or visible spectrometry or mass spectrometry for detection, or the like. Determining the levels of mRNA of the enzyme of interest may be accomplished using northern-blotting or RT-PCR techniques. Once plants have been regenerated, and progeny plants homozygous for the transgene have been obtained, plants will have a stable phenotype that will be observed in similar seeds in later generations.
[0200]Preferred Methods
[0201]The present invention also includes methods for altering stalk mechanical strength in a plant; methods for evaluating stalk mechanical strength in a plant; methods for evaluating cellulose content in plant; methods for altering cell wall cellulose content and/or growth rate in a plant, methods for conferring male sterility in a plant, and methods for reducing plant height and/or organ size in a plant. Preferably, the plant is a monocotyledonous or dicotyledonous plant, more preferably, a maize or soybean plant, even more preferably a maize plant. The plant may also be sunflower, sorghum, canola, wheat, alfalfa, cotton, rice, barley or millet.
[0202]A preferred method for altering (preferably increasing) stalk mechanical strength of a plant comprises (a) introducing a recombinant DNA construct into a regenerable plant cell to produce a transformed plant cell, the recombinant DNA construct comprising at least one regulatory element (preferably, a promoter that is functional in a plant) operably linked to (i) a nucleotide sequence encoding a polypeptide associated with stalk mechanical strength, wherein said polypeptide has an amino acid sequence of at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity, or any other integer in between 80% and 100%, based on the Clustal V method of alignment, when compared to SEQ ID NOs:4, 6, 8, 10, 12, 14, 16 or 18, or (ii) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary; and (b) regenerating a transgenic plant from said transformed plant cell, wherein said transgenic plant comprises in its genome said recombinant DNA construct and wherein said transgenic plant exhibits an alteration (preferably an increase) in stalk mechanical strength, when compared to a control plant not comprising said recombinant DNA construct.
[0203]A preferred method of evaluating stalk mechanical strength in a plant comprises (a) introducing into a regenerable plant cell a recombinant DNA construct to produce transformed plant cells, said recombinant DNA construct comprising a promoter that is functional in a plant operably linked to (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:4, 6, 8, 10, 12, 14, 16, and 18, or (ii) a full-length complement of said polynucleotide of (a)(i); (b) regenerating a transgenic plant from said transformed plant cell; and (c) evaluating said transgenic plant for stalk mechanical strength. This method may further comprise (d) obtaining a progeny plant derived from said transgenic plant; and (e) evaluating said progeny plant for stalk mechanical strength.
[0204]Another preferred method of evaluating stalk mechanical strength in a plant comprises (a) introducing into a regenerable plant cell a recombinant DNA construct to produce transformed plant cells, said recombinant DNA construct comprising a promoter that is functional in a plant operably linked to (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:4, 6, 8, 10, 12, 14, 16, and 18, or (b) a full-length complement of said polynucleotide of (a)(i); (b) regenerating a transgenic plant from said transformed plant cell; (c) obtaining a progeny plant derived from said transgenic plant; and (d) evaluating said progeny plant for stalk mechanical strength.
[0205]A preferred method of evaluating cellulose content in a plant, comprises (a) introducing into a regenerable plant cell a recombinant DNA construct to produce transformed plant cells, said recombinant DNA construct comprising a polynucleotide operably linked to a promoter that is functional in a plant, wherein said polynucleotide encodes a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:4, 6, 8, 10, 12, 14, 16, and 18; (b) regenerating a transgenic plant from said transformed plant cell; and (c) evaluating said transgenic plant for cellulose content. This method may further comprise (d) obtaining a progeny plant derived from said transgenic plant; and (e) evaluating said progeny plant for cellulose content.
[0206]Another preferred method of evaluating cellulose content in a plant comprises (a) introducing into a regenerable plant cell a recombinant DNA construct to produce transformed plant cells, said recombinant DNA construct comprising a polynucleotide operably linked to a promoter that is functional in a plant, wherein said polynucleotide encodes a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:4, 6, 8, 10, 12, 14, 16, and 18; (b) regenerating a transgenic plant from said transformed plant cell; (c) obtaining a progeny plant derived from said transgenic plant; and (d) evaluating said progeny plant for cellulose content.
[0207]A preferred method for selecting a plant with altered cellulose content comprises (a) obtaining any plant of the present invention (such as any of the preferred embodiments discussed above); (b) evaluating the plant obtained in step (a) for cellulose content; and (c) selecting the evaluated plant of step (b) when its cellulose content is altered when compared to a control plant. Preferably, the evaluated plant is selected when its cellulose content is increased, even more preferably, when the cellulose content is at least 35%, 40%, 45%, 50%, 55%, or 60% and/or when the cellulose dry matter content is at least 100 mg/cm, 200 mg/cm, 300 mg/cm, 400 mg/cm, or 500 mg/cm. Preferred methods for measuring cellose content are set forth herein in Example 10.
[0208]A preferred method for altering (preferably increasing) cell wall cellulose content and/or for altering (preferably enhancing) growth rate in a plant comprises integrating (e.g., through transgenic techniques or a combination of transgenic techniques and traditional breeding) into the genome of a plant one or more recombinant DNA constructs such that the co-expression is obtained of (a) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16, and 18; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15, and 17; and (iii) a full-length complement of the polynucleotide of (a)(i) or (a)(ii); and (b) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 81% sequence identity, or any integer up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, and 42; (ii) a polynucleotide having a nucleic acid sequence of at least 61% sequence identity, or any integer up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:21, 23, 25, 27, 29, 31, 33, 35, 37, 39, and 41; and (iii) a full-length complement of the polynucleotide of (b)(i) or (b)(ii).
[0209]Preferably, a method for increasing cell wall cellulose content in a plant comprises integrating into the genome of a plant one or more recombinant DNA constructs such that the co-expression is obtained of (a) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:2; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:1; and (iii) a full-length complement of the polynucleotide of (a)(i) or (a)(ii); and (b) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:38, 40, and 42; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:37, 39, and 41; and (iii) a full-length complement of the polynucleotide of (b)(i) or (b)(ii).
[0210]Preferably, a method for enhancing plant growth rate comprises integrating into the genome of a plant one or more recombinant DNA constructs such that the co-expression is obtained of (a) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:6; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:5; and (iii) a full-length complement of the polynucleotide of (a)(i) or (a)(ii); and (b) at least one isolated polynucleotide selected from the group consisting of (i) a polynucleotide encoding a polypeptide having an amino acid sequence of at least 80% sequence identity, or any integer from 81% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:20, 32, and 34; (ii) a polynucleotide having a nucleic acid sequence of at least 60% sequence identity, or any integer from 61% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NOs:19, 31, and 33; and (iii) a full-length complement of the polynucleotide of (b)(i) or (b)(ii).
[0211]A preferred method of conferring male sterility in a plant comprises: (a) introducing into a regenerable plant cell a suppression DNA construct comprising a promoter functional in a plant operably linked to (i) all or part of (A) a nucleic acid sequence encoding a polypeptide having an amino acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:10, or (B) a full complement of the nucleic acid sequence of (i)(A); or (ii) a region derived from all or part of a sense strand or antisense strand of a target gene of interest, said region having a nucleic acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to said all or part of a sense strand or antisense strand from which said region is derived, and wherein said target gene of interest encodes a Bk2L5 polypeptide; and (b) regenerating a transgenic plant from said transformed plant cell, wherein said transgenic plant comprises in its genome said suppression DNA construct and wherein said transgenic plant exhibits reduced plant height and/or reduced organ size when compared to a control plant not comprising said suppression DNA construct. The method may further comprise: (c) obtaining a progeny plant derived from said transgenic plant, wherein said progeny plant comprises in its genome the suppression DNA construct.
[0212]A preferred method of reducing plant height and/or reducing organ size in a plant comprises: (a) introducing into a regenerable plant cell a suppression DNA construct comprising a promoter functional in a plant operably linked to (i) all or part of (A) a nucleic acid sequence encoding a polypeptide having an amino acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity sequence identity, based on the Clustal V method of alignment, when compared to SEQ ID NO:6, or (B) a full complement of the nucleic acid sequence of (i)(A); or (ii) a region derived from all or part of a sense strand or antisense strand of a target gene of interest, said region having a nucleic acid sequence of at least 50% sequence identity, or any integer from 51% up to and including 100% sequence identity, based on the Clustal V method of alignment, when compared to said all or part of a sense strand or antisense strand from which said region is derived, and wherein said target gene of interest encodes a Bk2L3 polypeptide; and (b) regenerating a transgenic plant from said transformed plant cell, wherein said transgenic plant comprises in its genome said suppression DNA construct and wherein said transgenic plant exhibits reduced plant height and/or reduced organ size when compared to a control plant not comprising said suppression DNA construct. The method may further comprise: (c) obtaining a progeny plant derived from said transgenic plant, wherein said progeny plant comprises in its genome the suppression DNA construct.
[0213]The isolated nucleic acids and proteins and any embodiments of the present invention can be used over a broad range of plant types, particularly monocots such as the species of the Family Graminiae including Sorghum bicolor and Zea mays. The isolated nucleic acid and proteins of the present invention can also be used in species from the genera: Cucurbita, Rosa, Vitis, Juglans, Fragaria, Lotus, Medicago, Onobrychis, Trifolium, Trigonella, Vigna, Citrus, Linum, Geranium, Manihot, Daucus, Arabidopsis, Brassica, Raphanus, Sinapis, Atropa, Capsicum, Datura, Hyoscyamus, Lycopersicon, Nicotiana, Solanum, Petunia, Digitalis, Majorana, Ciahorium, Helianthus, Lactuca, Bromus, Asparagus, Antirrhinum, Heterocallis, Nemesis, Pelargonium, Panieum, Pennisetum, Ranunculus, Senecio, Salpiglossis, Cucumis, Browaalia, Glycine, Pisum, Phaseolus, Lolium, Oryza, Avena, Hordeum, Secale, Triticum, Bambusa, Dendrocalamus, and Melocanna.
EXAMPLES
[0214]The present invention is further illustrated in the following Examples, in which parts and percentages are by weight and degrees are Celsius, unless otherwise stated. It should be understood that these Examples, while indicating preferred embodiments of the invention, are given by way of illustration only. From the above discussion and these Examples, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions. Thus, various modifications of the invention in addition to those shown and described herein will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims.
Example 1
Characterization of Maize cDNA Encoding Bk2-Like Proteins
[0215]The maize brittle stalk 2 (bk2) phenotype was first reported in 1940 (Langham, MNL 14:21-22 (1940)), and was mapped by phenotype to chr9L between the markers umc95 and bnl7.13 around the 100 centiMorgan region (Howell et al., MNL 65:52-53 (1991)). Previously, clone csclc.pk005.k4:fis (SEQ ID NO:1) was shown to encode a BRITTLE STALK 2 polypeptide (SEQ ID NO:2) (International Application No. PCT/US2005/035450 which claims the benefit of U.S. Provisional Application No. 60/615,868, filed Oct. 6, 2004, the entire contents of which are herein incorporated by reference). Also disclosed were two other members of the Bk2 gene family (SEQ ID NOs:7 and 8 and SEQ ID NOs:13 and 14). In the instant disclosure these genes have been named as Bk2-like (Bk2L).
[0216]Search for additional maize cDNA sequences homologous at the nucleic acid and amino acid level to the maize BRITTLE STALK 2 (Bk2) sequence (SEQ ID NO:1) was conducted using BLASTN or TBLASTN algorithm provided by the National Center for Biotechnology Information (NCBI) against several databases, including, but not limited to, DuPont's internal proprietary database (Basic Local Alignment Search Tool; Altschul et al., J. Mol. Biol. 215:403-410 (1993); Altschul et al., Nucleic Acids Res. 25:3389-3402 (1997)) and publicly available Maize Genomic Survey Sequences (GSS) and TIGR Maize genomic assemblies (The TIGR Gene Index Databases, The Institute for Genomic Research, Rockville, Md. 20850; Quackenbush et al., J. Nucleic Acids Res. 28(1):141-145 (2000)). Six new members of the Bk2 gene family were isolated (Bk2L1, Bk2L3, Bk2L5, Bk2L6, Bk2L8 and Bk2L9). Table 1 lists all the Bk2-like proteins disclosed in the instant specification, in addition to Bk2 itself.
TABLE-US-00001 TABLE 1 Brittle Stalk 2-like Proteins SEQ ID NO: Protein Nucleotide Amino Acid Bk2 1 2 Bk2L1 3 4 Bk2L3 5 6 Bk2L4 7 8 Bk2L5 9 10 Bk2L6 11 12 Bk2L7 13 14 Bk2L8 15 16 Bk2L9 17 18
[0217]FIGS. 1A-1F show a Clustal V alignment, using default parameters, of the amino acid sequences reported in Table 1. FIG. 2 is a chart setting forth a comparison of the percent identity (and percent divergence in the lower half triangle), using the Clustal V alignment method, between the nine amino acid sequences shown in FIGS. 1A-1F.
[0218]The possible function of the polypeptide encoded by each cDNA was further identified by conducting BLAST (Basic Local Alignment Search Tool; Altschul, S. F., et al., J. Mol. Biol. 215:403-410 (1993)) searches of the ESTs against public databases. The searches were conducted for similarity to sequences contained in the BLAST "nr" database (comprising all non-redundant GenBank CDS translations, sequences derived from the 3-dimensional structure Brookhaven Protein Data Bank, the last major release of the SWISS-PROT protein sequence database, EMBL, and DDBJ databases). The sequences were analyzed for similarity using the BLASTN algorithm provided by the National Center for Biotechnology Information (NCBI). The DNA sequences were translated in all reading frames and compared for similarity to all publicly available protein sequences contained in the "nr" database using the BLASTX algorithm (Gish, W. and States, D. J., Nature Genetics 3:266-272 (1993)) provided by the NCBI. Shown in Table 2 are the "Score" results obtained for the amino acid sequences of the entire Bk2-like proteins encoded by the entire cDNA inserts comprising the indicated cDNA clones. The data in Table 2 also presents the results obtained for the calculation of the percent identity of the amino acid sequences set forth in SEQ ID NOs:2, 4, 6, 8, 10, 12, 14, 16 and 18, with the sequences identified in the NCBI General Identifier No. column.
TABLE-US-00002 TABLE 2 BLAST Results for Sequences Encoding Polypeptides Homologous to Bk2-like Proteins NCBI General Gene Identifier No. Percent (SEQ ID NO:) (Accession No.) Score (bits) Identity Bk2L1 NCBI GI 34733385 1266 100% (SEQ ID NO: 3) (AAQ81633.1) Bk2L3 NCBI GI 30090026 868 95% (SEQ ID NO: 5) (AAO17706.1) Bk2L4 NCBI GI 30090026 922 97% (SEQ ID NO: 7) (AAO17706.1) Bk2L5 NCBI GI 52076665 1079 79% (SEQ ID NO: 9) (BAD45565.1) Bk2L6 NCBI GI 50939113 742 81% (SEQ ID NO: 11) (XP_479084.1) Bk2L7 NCBI GI 50939113 751 82% (SEQ ID NO: 13) (XP_479085.1) Bk2L8 NCBI GI 34898176 838 65% (SEQ ID NO: 15) (NP_910434.1) Bk2L9 NCBI GI 50927043 597 63% (SEQ ID NO: 17) (XP_473354.1)
[0219]FIG. 6 shows the phylogenetic analysis of the Bk2L proteins from maize, BC1L proteins from rice, and COBL proteins from Arabidopsis (NCBI Accession Nos. are in parenthesis). The numbers along the branches are the bootstrap values obtained from a heuristic search over 5,000 replications. The bootstrap values for only the monophyletic groups that were supported >50% of the time are shown. The branch lengths are proportional to the inferred amino acid differences.
Example 2
Gene Expression Analysis of Bk2-Like Proteins
[0220]The tissue specificity of expression of the Bk2-like gene family disclosed in Table 1 was examined using Solexa's Massively Parallel Signature Sequencing (MPSS®) technology (see Table 3) (Brenner et al., Nat. Biotechnol. 18:630-634 (2000); Brenner et al., Proc. Natl. Acad. Sci. U.S.A. 97:1665-1670 (2000)). MPSS® involves the generation of seventeen base signature tags from mRNA samples that have been reverse transcribed. The tags are simultaneously sequenced and assigned to genes or ESTs. The abundance of these tags is given a number value that is normalized to parts per million (PPM) which then allows the tag expression, or tag abundance, to be compared across different tissues. Thus, the MPSS® platform can be used to determine the expression pattern of a particular gene and its expression level in different tissues. The numbers are averages over multiple libraries for each tissue listed in the second column.
TABLE-US-00003 TABLE 3 Expression in PPM of the Bk2 Gene Family in Maize Tissue Lib. # Bk2 Bk2L1 Bk2L3 Bk2L4 Bk2L5 Bk2L6 Bk2L7 Bk2L8 Bk2L9 anther 3 1 73 49 51 0 0 0 9 1 ear 17 0 48 17 30 0 0 0 0 0 embryo 10 0 18 61 18 0 0 0 0 3 endosperm 26 0 18 48 23 0 4 1 3 0 husk 1 75 68 490 16 0 0 39 0 0 kernel 5 2 86 103 51 0 1 1 15 1 leaf 46 17 20 87 10 0 0 0 32 0 meristem 20 2 60 81 19 0 0 0 2 0 pericarp 6 4 16 290 54 0 0 0 2 0 pollen 2 0 6 2 13 794 0 0 0 0 root 43 52 69 263 14 0 0 2 8 0 seedling 7 8 16 72 21 0 0 0 8 0 silk 9 0 36 69 47 29 0 2 0 0 spikelet 12 17 86 205 111 0 0 12 0 0 stalk 15 172 48 474 15 0 0 8 14 0 tassel 2 4 72 53 62 0 0 16 0 0 vascular 2 182 56 117 11 0 0 0 7 0 bundles whorl 7 152 9 126 33 0 0 0 2 2
[0221]Bk2 (Table 3, column 3 and FIG. 3) is expressed in husk, leaf, root, stalk and isolated vascular bundles, but not in the kernel, meristem, pollen or silk tissues. This expression pattern is consistent with the role of the Bk2 gene in secondary wall formation as all the tissues it is expressed in contain at least some lignified cells. The correlation coefficient analysis of the expression level of Bk2 with the expression levels of the twelve maize CesA genes is shown in FIG. 4 (also see FIG. 5A, column 2). The expression pattern of the Bk2 gene is very similar to that of the previously disclosed secondary wall-forming CesA genes, CesA10, 11 and 12 (see FIG. 5 of U.S. Pat. No. 6,930,225, granted Aug. 16, 2005, the entire contents of which are herein incorporated by reference). More specifically, Bk2 shows a higher correlation coefficient, approximately >0.8, with each of the maize CesA10, 11, and 12 genes than with any other gene in this class. Since the three CesA genes are also co-expressed, it is likely that their corresponding proteins form a functional complex along with the Bk2 protein. Table 4 lists all the primary and secondary wall-forming CesA proteins known to date (U.S. Pat. No. 6,930,225, supra; U.S. Pat. No. 6,803,498, granted Oct. 12, 2004, the entire contents of which are herein incorporated by reference). The maize CesA10, 11, and 12 genes and their orthologs from Arabidopsis and rice have been implicated in secondary wall formation (Tanaka et al., Plant Physiol. 133:73-83 (2003); Taylor et al., Proc. Natl. Acad. Sci. U.S.A. 100:1450-1455 (2003); Appenzeller et al., Cellulose 11:287-299 (2004)). The co-expression of the Bk2 and secondary wall-forming CesA genes supports a role for Bk2 in secondary wall formation in maize.
TABLE-US-00004 TABLE 4 Primary and Secondary Wall-forming CesA Proteins SEQ ID NO: Protein (Nucleotide) (Amino Acid) CesA1 19 20 CesA2 21 22 CesA3 23 24 CesA4 25 26 CesA5 27 28 CesA6 29 30 CesA7 31 32 CesA8 33 34 CesA9 35 36 CesA10 37 38 CesA11 39 40 CesA12 41 42
[0222]Another Bk2L gene that shows correlated expression with CesA genes is Bk2L3. The expression pattern of Bk2L3, is very similar to the CesA genes that were reported previously to be involved in primary wall formation (Holland et al., Plant Physiol. 123:1313-1323 (2000); Dhugga, Curr. Opin. Plant Biol. 4:488-493 (2001); Appenzeller et al., Cellulose 11:287-299 (2004)). Three genes in particular, CesA1, 7 and 8 appear to likely form a functional cellulose synthase complex for primary wall formation. The expression of the Bk2L3 gene is highly correlated with these three CesA genes that it appears that, analogous to the secondary wall cellulose synthase complex consisting of three CesA proteins and a Bk2 protein, these four proteins may form a functional cellulose synthase complex for primary wall formation.
[0223]Bk2L5 is expressed only in pollen. Some expression in silk most likely results from the pollen tube growing through it. Bk2L8 appears like is leaf-preferred and Bk2L6 is endosperm-specific.
[0224]Correlation among the expression level of all the different Bk2 and CesA genes from maize as studied from Solexa MPSS® is shown in FIGS. 5A and 5B.
Example 3
Prophetic Example
Engineering Increased Stalk Strength by Overexpression of Maize Bk2-Like Genes Under a Strong, Stalk-Specific Promoter
[0225]A chimeric transgene is constructed to directly overexpress the Bk2 gene/polypeptide in a tissue specific manner. The transgene construct comprises a maize cDNA encoding Bk2L3 and/or Bk2L6 (e.g., SEQ ID NO:5 or SEQ ID NO:11) operably linked to the promoter from the alfalfa stalk-specific S2A gene (Abrahams et al., Plant Mol. Biol. 27:513-528 (1995)). The DNA containing the Bk2L3 or Bk2L6 ORF is then fused to the S2A promoter on the 5' end and pinII terminator on the 3' end to produce an expression cassette as illustrated in FIG. 3. The construct is then linked to a selectable marker cassette containing a bar gene driven by CaMV 35S promoter and a pinII terminator. It is appreciated that one skilled in the art could employ different promoters, 5' end sequences and/or 3' end sequences to achieve comparable expression results. Transgenic maize plants are produced by transforming immature maize embryos with this expression cassette using the Agrobacterium-based transformation method by Zhao (U.S. Pat. No. 5,981,840, issued Nov. 9, 1999; the contents of which are hereby incorporated by reference). While the method below is described for the transformation of maize plants with the S2A promoter-Bk2L3 (or Bk2L6) expression cassette, those of ordinary skill in the art recognize that this method can be used to produce transformed maize plants with any nucleotide construct or expression cassette that comprises a promoter linked to maize Bk2L3 (or Bk2L6) gene for expression in a plant.
[0226]Immature embryos are isolated from maize and the embryos contacted with a suspension of Agrobacterium, where the bacteria are capable of transferring the S2A promoter-Bk2L3 (or Bk2L6) expression cassette (illustrated above) to at least one cell of at least one of the immature embryos (step 1: the infection step). In this step, the immature embryos are immersed in an Agrobacterium suspension for the initiation of inoculation. The embryos are co-cultured for a time with the Agrobacterium (step 2: the co-cultivation step). The immature embryos are cultured on solid medium following the infection step. Following this co-cultivation period an optional "resting" step is included. In this resting step, the embryos are incubated in the presence of at least one antibiotic known to inhibit the growth of Agrobacterium without the addition of a selective agent for plant transformants (step 3: resting step). The immature embryos are cultured on solid medium with antibiotic, but without a selecting agent, for elimination of Agrobacterium and for a resting phase for the infected cells. Next, inoculated embryos are cultured on medium containing a selective agent and growing transformed calli are recovered (step 4: the selection step). Preferably, the immature embryos are cultured on solid medium with a selective agent resulting in the selective growth of transformed cells. The resulting calli are then regenerated into plants by culturing the calli on solid, selective medium (step 5: the regeneration step).
Example 4
Prophetic Example
Engineering Increased Stalk Strength by Transgenic Expression of Maize Bk2-Like Genes with an Enhancer Element in the Promoter Region Under a Strong, Stalk-Specific Promoter
[0227]The expression of the Bk2L3 (or Bk2L6) gene is increased by placing a heterologous enhancer element in the promoter region of the native Bk2L3 (or Bk2L6) gene. An expression cassette is constructed comprising an enhancer element such as CaMV 35S fused to the native promoter of Bk2L3 (or Bk2L6) and the full-length cDNA. Transgenic maize plants can then be produced by transforming immature maize embryos with this expression cassette as described in Example #3.
Example 5
Prophetic Example
Engineering Increased Stalk Strength by Overexpression of Maize Bk2-Like and CesA Genes
[0228]Whereas the secondary wall-forming genes mainly affect the mechanical strength of the plant tissues and not the morphological phenotype, the primary wall-forming genes can affect plant growth rate and thus their modulation can be employed to increase the rate of growth. The maize genes CesA1, 7, and 8 were previously shown to be co-expressed across multiple tissues, suggesting that they might form a functional enzyme complex. Bk2L3 is co-expressed with these three CesA genes, strongly suggesting that the protein products of all of these four genes form a functional enzyme complex. Simultaneous over-expression of these four genes as a single multi-gene construct or as separate constructs containing different combinations of these genes in maize driven by different promoters, preferably by the promoters of genes whole expression is associated with cell elongation, can be employed to produce transgenic plants with enhanced growth rate. Any of the other Bk2L genes can also be used in combination with the mentioned three CesA genes as described above to produce transgenic plants with enhanced growth rate.
Example 6
Prophetic Example
Engineering Increased Stalk Strength by Overexpression of Maize Bk2-Like and CesA Genes
[0229]Aside from contributing to mechanical strength, secondary wall accounts for a majority of the biomass in plants. Whereas mechanical strength has applications in reducing in crop lodging, quality and amount biomass are important for many other applications, including ethanol production. The Bk2 gene along with the maize CesA10, 11, and 12 genes offers an avenue to increase the ratio of cellulose in the cell wall. The efficiency of ethanol production is directly related to the amount of cellulose in the biomass. Replacement of lignin with cellulose will also be useful in silage digestibility.
[0230]The Bk2 gene can be co-expressed with the CesA10, 11, and 12 genes as described in Example 5 for the primary wall-forming genes but under the control of secondary wall-specific promoters to produce transgenic plants with improved stalk strength and biomass quality.
Example 7
Prophetic Example
Engineering Down-regulation of Maize Bk2-Like Genes
[0231]Since primary wall forming CesA genes contribute to cell expansion, their limited down-regulation can be employed to reduce plant height or organ size. In particular, the expression of the Bk2L3 gene is highly correlated with the primary wall-forming CesA genes. Whereas the overexpression of all the members of a functional enzyme complex may be required to increased enzyme activity, down-regulation of only one member may be sufficient to reduce activity. The down-regulation of Bk2L3, for example (and/or Bk2L5 for male sterility), can be accomplished by any of the technologies of co-suppression, RNAi, antisense RNA, or micro RNA resulting in dwarf transgenic plants. Height reduction has applications in some crop plants where harvest index is low and needs to be increased. Modern wheat and rice varieties, for example, are considerably shorter than their older counterparts. The ability to reduce plant height was mainly the cause of green revolution in each of these crops.
Example 8
Prophetic Example
Expression of Recombinant DNA Constructs in Dicot Cells Under a Strong, Stalk-Specific Promoter
[0232]An expression cassette composed of the promoter from the alfalfa stalk-specific S2A gene (Abrahams et al., Plant Mol. Biol. 27:513-528 (1995)) 5-prime to the cDNA fragment can be constructed and be used for expression of the instant polypeptides in transformed soybean. The pinII terminator can be placed 3-prime to the cDNA fragment. Such construct may be used to overexpress the Bk2-like genes. It is realized that one skilled in the art could employ different promoters and/or 3-prime end sequences to achieve comparable expression results.
[0233]The cDNA fragment of this gene may be generated by polymerase chain reaction (PCR) of the cDNA clone using appropriate oligonucleotide primers. Cloning sites can be incorporated into the oligonucleotides to provide proper orientation of the DNA fragment when inserted into the expression vector. Amplification is then performed as described above, and the isolated fragment is inserted into a pUC18 vector carrying the seed expression cassette.
[0234]Soybean embryos may then be transformed with the expression vector comprising sequences encoding the instant polypeptides. To induce somatic embryos, cotyledons, 3-5 mm in length dissected from surface sterilized, immature seeds of the soybean cultivar A2872, can be cultured in the light or dark at 26° C. on an appropriate agar medium for 6-10 weeks. Somatic embryos which produce secondary embryos are then excised and placed into a suitable liquid medium. After repeated selection for clusters of somatic embryos which multiplied as early, globular staged embryos, the suspensions are maintained as described below.
[0235]Soybean embryogenic suspension cultures can be maintained in 35 mL liquid media on a rotary shaker, 150 rpm, at 26° C. with florescent lights on a 16:8 hour day/night schedule. Cultures are subcultured every two weeks by inoculating approximately 35 mg of tissue into 35 mL of liquid medium.
[0236]Soybean embryogenic suspension cultures may then be transformed by the method of particle gun bombardment (Klein et al. (1987) Nature (London) 327:70-73, U.S. Pat. No. 4,945,050). A DuPont Biolistic® PDS1000/HE instrument (helium retrofit) can be used for these transformations.
[0237]A selectable marker gene which can be used to facilitate soybean transformation is a chimeric gene composed of the 35S promoter from cauliflower mosaic virus (Odell et al. (1985) Nature 313:810-812), the hygromycin phosphotransferase gene from plasmid pJR225 (from E. coli; Gritz et al. (1983) Gene 25:179-188) and the 3' region of the nopaline synthase gene from the T-DNA of the Ti plasmid of Agrobacterium tumefaciens. The seed expression cassette comprising the phaseolin 5' region, the fragment encoding the instant polypeptides and the phaseolin 3' region can be isolated as a restriction fragment. This fragment can then be inserted into a unique restriction site of the vector carrying the marker gene.
[0238]To 50 μL of a 60 mg/mL 1 μm gold particle suspension is added (in order): 5 μL DNA (1 μg/μL), 20 μL spermidine (0.1 M), and 50 μL CaCl2 (2.5 M). The particle preparation is then agitated for three minutes, spun in a microfuge for 10 seconds and the supernatant removed. The DNA-coated particles are then washed once in 400 μL 70% ethanol and resuspended in 40 μL of anhydrous ethanol. The DNA/particle suspension can be sonicated three times for one second each. Five μL of the DNA-coated gold particles are then loaded on each macro carrier disk.
[0239]Approximately 300-400 mg of a two-week-old suspension culture is placed in an empty 60×15 mm petri dish and the residual liquid removed from the tissue with a pipette. For each transformation experiment, approximately 5-10 plates of tissue are normally bombarded. Membrane rupture pressure is set at 1100 psi and the chamber is evacuated to a vacuum of 28 inches mercury. The tissue is placed approximately 3.5 inches away from the retaining screen and bombarded three times. Following bombardment, the tissue can be divided in half and placed back into liquid and cultured as described above.
[0240]Five to seven days post bombardment, the liquid media may be exchanged with fresh media, and eleven to twelve days post bombardment with fresh media containing 50 mg/mL hygromycin. This selective media can be refreshed weekly. Seven to eight weeks post bombardment, green, transformed tissue may be observed growing from untransformed, necrotic embryogenic clusters. Isolated green tissue is removed and inoculated into individual flasks to generate new, clonally propagated, transformed embryogenic suspension cultures. Each new line may be treated as an independent transformation event. These suspensions can then be subcultured and maintained as clusters of immature embryos or regenerated into whole plants by maturation and germination of individual somatic embryos.
Example 9
Prophetic Example
Expression of Recombinant DNA Constructs in Microbial Cells Under a Strong, Stalk-Specific Promoter
[0241]The cDNAs (SEQ ID NOs:1, 3, 5, 7, 9, 11, 13, 15 or 17) encoding the instant BRITTLE STALK 2-like polypeptides can be inserted into the T7 E. coli expression vector pBT430. This vector is a derivative of pET-3a (Rosenberg et al. (1987) Gene 56:125-135) which employs the bacteriophage T7 RNA polymerase/T7 promoter system. Plasmid pBT430 is constructed by first destroying the EcoRI and HindIII sites in pET-3a at their original positions. An oligonucleotide adaptor containing EcoRI and Hind III sites is inserted at the BamHI site of pET-3a. This creates pET-3aM with additional unique cloning sites for insertion of genes into the expression vector. Then, the NdeI site at the position of translation initiation is converted to an NcoI site using oligonucleotide-directed mutagenesis. The DNA sequence of pET-3aM in this region, 5'-CATATGG, is converted to 5'-CCCATGG in pBT430.
[0242]Plasmid DNA containing a cDNA may be appropriately digested to release a nucleic acid fragment encoding the protein. This fragment may then be purified on a 1% low melting agarose gel. Buffer and agarose contain 10 μg/ml ethidium bromide for visualization of the DNA fragment. The fragment can then be purified from the agarose gel by digestion with GELase® (Epicentre Technologies, Madison, Wis.) according to the manufacturer's instructions, ethanol precipitated, dried and resuspended in 20 μL of water. Appropriate oligonucleotide adapters may be ligated to the fragment using T4 DNA ligase (New England Biolabs (NEB), Beverly, Mass.). The fragment containing the ligated adapters can be purified from the excess adapters using low melting agarose as described above. The vector pBT430 is digested, dephosphorylated with alkaline phosphatase (NEB) and deproteinized with phenol/chloroform as described above. The prepared vector pBT430 and fragment can then be ligated at 16° C. for 15 hours followed by transformation into DH5 electrocompetent cells (GIBCO BRL). Transformants can be selected on agar plates containing LB media and 100 μg/mL ampicillin. Transformants containing the gene encoding the instant polypeptides are then screened for the correct orientation with respect to the T7 promoter by restriction enzyme analysis.
[0243]For high level expression, a plasmid clone with the cDNA insert in the correct orientation relative to the T7 promoter can be transformed into E. coli strain BL21 (DE3) (Studier et al. (1986) J. Mol. Biol. 189:113-130). Cultures are grown in LB medium containing ampicillin (100 mg/L) at 25° C. At an optical density at 600 nm of approximately 1, IPTG (isopropylthio-β-galactoside, the inducer) can be added to a final concentration of 0.4 mM and incubation can be continued for 3 h at 25° C. Cells are then harvested by centrifugation and re-suspended in 50 μL of 50 mM Tris-HCl at pH 8.0 containing 0.1 mM DTT and 0.2 mM phenyl methylsulfonyl fluoride. A small amount of 1 mm glass beads can be added and the mixture sonicated 3 times for about 5 seconds each time with a microprobe sonicator. The mixture is centrifuged and the protein concentration of the supernatant determined. One μg of protein from the soluble fraction of the culture can be separated by SDS-polyacrylamide gel electrophoresis. Gels can be observed for protein bands migrating at the expected molecular weight.
Example 10
Characteristics of the Stalk Tissue of the Wildtype (Bk2) and of the Brittle Stalk (bk2-ref) Mutant of Maize
[0244]The maize stock containing the reference allele of bk2 (bk2-ref) was obtained from the Maize Genetics COOP Stock Center (USDA/ARS & Crop Sciences/UIUC, S-123 Turner Hall, 1102 S. Goodwin Avenue, Urbana, Ill. 61801-4798). Three greenhouse-grown plants each of the bk2-ref/bk2-ref and its wildtype sibling, Bk2/bk2-ref, both derived from seeds obtained from the same selfed ear, were evaluated for different traits approximately two weeks after flowering. Three internodes below the ear (internodes 3, 4, and 5, numbered from the ear node) were subjected to a three-point flexural test using a model 4411 Instron electromechanical testing device (Instron Corp., Canton, Mass.). The span width between the anchor points was 20 cm. The anvil was vertically driven at a constant speed of 20 cm/min against the internodal zone ˜3 cm above the node on a horizontally placed stalk until it collapsed or snapped. The maximum load to break was used as a measure of strength to differentiate the internodes and stalks.
[0245]Total dry matter was measured in the stalk portion below the ear node. Structural dry matter and cellulose contents were determined in duplicates on each of the three plants from the third and fourth internodes below the ear node by boiling the powdered stalk material twice with buffer (25 mM MOPS, pH 7) for 30 minutes. The remaining material was suspended in methanol/chloroform (3/1, v/v) for 1 hour, dried and weighed. Crystalline cellulose was determined by the Updegraff method (Updegraff, Anal. Biochem. 32:120-124 (1969)). Briefly, ground stalk material was place in a boiling water bath in an 8:2:1 mixture of acetic acid:water:nitric acid for 1 hour, the crystalline material washed three times with water and then with 95% ethanol followed by drying in a Speedvac. Klason lignin was determined by incubating the ground stalk material with 72% (w/w) sulfuric acid for 1 hour, washing twice with a 1:20 dilution of 72% sulfuric acid in water, heating at 65° C. for 30 minutes, washing once with water, and drying the residue at 80° C. overnight. Sugar composition was determined as described in (Appenzeller et al., Cellulose 11:287-299 (2004)).
[0246]In summary, reduction in mechanical strength in the stalk tissue was highly correlated with a reduction in the amount of cellulose and an uneven deposition of secondary cell wall material in the subepidermal and perivascular sclerenchyma fibers. Lower amount of cellulose and thinner walls of the mutant were reflected in reduced dry matter content per unit length of the stalk.
TABLE-US-00005 TABLE 5 Measurement of Stalk Composition and Mechanical Strength Trait Wildtype bk2-ref ear height (cm) 102.00 ± 8.8 106.33 ± 11.8 stalk diameter (mm) 23.84 ± 0.27 23.40 ± 0.46 stalk dry mass (g) 89.43 ± 3.39 62.08 ± 8.46 moisture (%) 79.20 ± 0.21 84.87 ± 1.04 dry matter (g/cm) 0.68 ± 0.04 0.43 ± 0.07 displacement to break (mm) 11.83 ± 0.46 6.51 ± 1.10 load to break (kg) 23.68 ± 2.25 9.04 ± 2.66 insoluble dry matter (%) 51.57 ± 1.00 45.20 ± 1.69 cellulose (%) 33.30 ± 0.56 23.76 ± 0.68 lignin (%) 9.07 ± 0.21 10.28 ± 0.63 remainder cell wall (%) 9.20 ± 1.68 11.16 ± 2.03 cellulose (g/cm) 0.24 ± 0.114 0.11 ± 0.019
[0247]region is derived, and wherein said target gene of interest: encodes a Bk2L5 polypeptide,
[0248]and wherein said plant exhibits male sterility when compared to a control plant not comprising said suppression DNA construct.
Sequence CWU
1
4211784DNAZea mays 1gatcggagct tgtgctgcta ctgctactat accagcgcta gctagcagca
gccgccggcc 60ggctcgcgca agctaaggaa gggtcgacat gacgatgggg ctccgcgtcc
gcgactcctc 120cgcgctgctg gctctggccg tcgcgctcgc ctgctgctcc gttgcagtgg
tggcctacga 180ccccctggac ccgaacggca acatcaccat caagtgggac gtgatctcgt
ggacgcccga 240cgggtacgtg gcgatggtga cgatgagcaa ctaccagatg taccggcaca
tcatggcgcc 300cgggtggacg ttggggtggt cgtgggccaa gaaggaggtg atctggtcca
tcgtgggggc 360gcaggccacg gagcaggggg actgctccaa gttcaagggc ggcatcccgc
actgctgcaa 420gcgcaccccg gccgtggtgg acctcctccc gggggtgccc tacaaccagc
agatcgccaa 480ctgctgcaag gccggcgtgg tgtcggcgta cgggcaggac ccggcggggt
ccgtctccgc 540gttccaggtc tccgtcggcc tggccggtac caccaacaag acggtgaagc
tgcccaggaa 600cttcacgctc atggggcccg ggctgggcta cacctgcggg cccgccgccg
tggtgccgtc 660caccgtgtac tggacgcccg accaccggcg ccggacgcag gcgctcatga
cgtggacggt 720gacctgcacc tactcgcagc agctggcgtc ccggtacccg tcctgctgcg
tctccttctc 780ctccttctac aacagcacca tcgtgccgtg cgcccggtgc gcgtgcggct
gcggcggcca 840cggcggccac gcgggtccgg gcggctgcat cgagggggac tccaagcgcg
cgctgtcggc 900cggggtgaac acgccgcgca aggacggcca ggcgctgctg cagtgcacgc
cgcacatgtg 960ccccatccgg gtgcactggc acgtcaagct caactacaag gactactggc
gcgccaagat 1020cgccatcacc aactacaact acaggatgaa ctacacgcag tggacgctgg
tggcgcagca 1080ccccaacctg gacaacgtca ccgaggtctt cagcttccag tacaagccgc
tgcaaccata 1140cgggagcatc aatgacactg gcatgttcta cgggctcaag ttctacaacg
actttctcat 1200ggaggccggc ccgttcggca acgtgcagtc ggaggtgctc atgcgcaagg
acgcaaggac 1260cttcaccttc agcatgggct gggcgttccc gcgcaagatc tacttcaacg
gcgacgagtg 1320caagatgccg ccgccggact cctaccccta cctgcccaac gccgcgcccg
tcgtcgcctc 1380gcagctggtc ctgtccgccg ccgcctcggc gttcctactg ttgctgctcc
tggtggcatg 1440accgtgaccg aaccaagggc aaggcctccg ttttgttttc ccgtctcgtc
ccgtgggcag 1500ggagcagact tcagtaggca gggcatttta tttggttttt ttgccaagga
ttcaacactt 1560gggttttcgt cagaggaaaa ctgtcgtgta tgtagtgtga gttgcaggtc
gtcggatccc 1620cacgtacaag acaatctttg gatctagaat atgcaaaacg tgaatcagca
cgccaggatc 1680atcgtctcct acaagattgg cagaaaaaaa atctcatgat gagtgatgtg
tcaacagacc 1740tatatatatg tgataatcac tggtttcaaa aaaaaaaaaa aaaa
17842450PRTZea mays 2Met Thr Met Gly Leu Arg Val Arg Asp Ser
Ser Ala Leu Leu Ala Leu1 5 10
15Ala Val Ala Leu Ala Cys Cys Ser Val Ala Val Val Ala Tyr Asp Pro
20 25 30Leu Asp Pro Asn Gly Asn
Ile Thr Ile Lys Trp Asp Val Ile Ser Trp 35 40
45Thr Pro Asp Gly Tyr Val Ala Met Val Thr Met Ser Asn Tyr
Gln Met 50 55 60Tyr Arg His Ile Met
Ala Pro Gly Trp Thr Leu Gly Trp Ser Trp Ala65 70
75 80Lys Lys Glu Val Ile Trp Ser Ile Val Gly
Ala Gln Ala Thr Glu Gln 85 90
95Gly Asp Cys Ser Lys Phe Lys Gly Gly Ile Pro His Cys Cys Lys Arg
100 105 110Thr Pro Ala Val Val
Asp Leu Leu Pro Gly Val Pro Tyr Asn Gln Gln 115
120 125Ile Ala Asn Cys Cys Lys Ala Gly Val Val Ser Ala
Tyr Gly Gln Asp 130 135 140Pro Ala Gly
Ser Val Ser Ala Phe Gln Val Ser Val Gly Leu Ala Gly145
150 155 160Thr Thr Asn Lys Thr Val Lys
Leu Pro Arg Asn Phe Thr Leu Met Gly 165
170 175Pro Gly Leu Gly Tyr Thr Cys Gly Pro Ala Ala Val
Val Pro Ser Thr 180 185 190Val
Tyr Trp Thr Pro Asp His Arg Arg Arg Thr Gln Ala Leu Met Thr 195
200 205Trp Thr Val Thr Cys Thr Tyr Ser Gln
Gln Leu Ala Ser Arg Tyr Pro 210 215
220Ser Cys Cys Val Ser Phe Ser Ser Phe Tyr Asn Ser Thr Ile Val Pro225
230 235 240Cys Ala Arg Cys
Ala Cys Gly Cys Gly Gly His Gly Gly His Ala Gly 245
250 255Pro Gly Gly Cys Ile Glu Gly Asp Ser Lys
Arg Ala Leu Ser Ala Gly 260 265
270Val Asn Thr Pro Arg Lys Asp Gly Gln Ala Leu Leu Gln Cys Thr Pro
275 280 285His Met Cys Pro Ile Arg Val
His Trp His Val Lys Leu Asn Tyr Lys 290 295
300Asp Tyr Trp Arg Ala Lys Ile Ala Ile Thr Asn Tyr Asn Tyr Arg
Met305 310 315 320Asn Tyr
Thr Gln Trp Thr Leu Val Ala Gln His Pro Asn Leu Asp Asn
325 330 335Val Thr Glu Val Phe Ser Phe
Gln Tyr Lys Pro Leu Gln Pro Tyr Gly 340 345
350Ser Ile Asn Asp Thr Gly Met Phe Tyr Gly Leu Lys Phe Tyr
Asn Asp 355 360 365Phe Leu Met Glu
Ala Gly Pro Phe Gly Asn Val Gln Ser Glu Val Leu 370
375 380Met Arg Lys Asp Ala Arg Thr Phe Thr Phe Ser Met
Gly Trp Ala Phe385 390 395
400Pro Arg Lys Ile Tyr Phe Asn Gly Asp Glu Cys Lys Met Pro Pro Pro
405 410 415Asp Ser Tyr Pro Tyr
Leu Pro Asn Ala Ala Pro Val Val Ala Ser Gln 420
425 430Leu Val Leu Ser Ala Ala Ala Ser Ala Phe Leu Leu
Leu Leu Leu Leu 435 440 445Val Ala
45033152DNAZea mays 3cctgaacctc tcctcggcac atgcgcgggc cccactttac
aagcgacagt agccccatcc 60gcagccgtgc acgctagaat cggacggctt gccgcgcatc
tcgcccgtcc gcggcgccgc 120tgcccgaacc gaaccctacc gtttcaacca ggcgccgcct
ctccgcgtcc gctgagtcac 180tgcctcctgg gcccggaccc acacgcccct cgtcaatcaa
catcaactca cgctcctcat 240catccctcca ctggaaactg gaccccctcg tcgtctcgtc
tcttttcctg ccggcgtcga 300ttcccactcc gttttgttaa aaaccgatcg ttttctccat
ttctttgtag ggactagtaa 360tagatagaca gcagagggag agacgacagg cgtagctagc
accagcactc aagctactac 420gcacgcacgc cgccgctccc ccagttcaaa cccaccaccc
cttccccctt catcttcctt 480tcccagctgt gcacgcgctt tccgatcgct tcatctacct
cgccaccgcg cttccgccca 540gccccagtca ccagtccacc gcgcccgcgc ccccgatcca
gcgatatggc tggctccgta 600gctccccacg ctgtggtcct cggtcttctc ctgctcgcgg
ggctcgcggc ggcgcagagg 660gcgacgacgc cggctgcggc ggcccccgcg cccgaccccg
gctgcaacgg catccagctg 720acctacaact tcgtggaccg caccaagatc cggcccttcg
tcagcgacaa gaacaagcag 780ccctacgcct tccgcgccaa cgtcaccgtg ctcaactccg
gcacccgccc gctcaagtcc 840tgggcggcac tcgtcacatt cggctacggc gagatcctcg
tcggcgtcga cggcgccgtg 900ctcacgggcg gcggcgacct gccgtacaac accacggagg
acgccggcaa cgccacctcg 960ttctccgggt acccgcatac agacctcctc acgcccatcg
ccaccgccgg ggacctgtcg 1020cagatccagg cctccgtcgg catcgtcggc acgctcttcg
ccgggcccgg cccgttcgtg 1080ccgctcccca ccgcgctgtc gctggacgac ccggcctacg
cgtgcccggc ggcgaccaac 1140gtcactgctc gggtgctgtc cacgtgctgc gtcctcacgc
cggaggccga ggccaacgcc 1200actgccatcg acgccaacac caccgacccg accaaggatt
tcctgccgcg cggcaccggc 1260gacctcgtca tcacctacga tgtgctccag gcctacccct
ccagctacct tgcgctcgtc 1320acgctcgaga acaacgccaa gctcggccgc ctcgacaact
ggcggctgtc gtgggagtgg 1380cggcgtgggg agttcatcta ctcaatgaaa ggagctcacc
catcagaggt ggacacctcg 1440ggctgtatct gtggggcgcc tgggcagtac taccagagcc
ttgatttttc gcaggtgctc 1500aattgtgacc gcaagccggt gatccttgac ctgcccctgt
cccggtacaa cgacactcag 1560attgggaaga ttgacaattg ctgcaggaat gggacaatct
tgcccaagtc catggacgag 1620gcacagtcga aatctgcgtt ccagatgcaa gttttcaaga
tgccaccaga cctgaaccgg 1680actaagctgt tcccccctgc taatttcaag atcgtgggtg
catcatcgct gaacccggac 1740tatgcctgtg gccagccggt gcctgtcagc ccaaccgcgt
tcccagaccc gagcgggctt 1800gactcgacga cgcttgctgt ggcaacatgg caggtggtgt
gcaacattac cacgacaaag 1860ggggccaagc ccaagtgttg tgtgaccttc tcggcgtact
acaacgactc agtgatcccc 1920tgcagcacct gcgcttgtgg gtgccctgca aacaggcgag
ggccaacgtg cagcaccacc 1980gcacaatcca tgctgctgcc accggaggcg ctgcttgtgc
cattcgacaa ccggtcacag 2040aaggcgttgg cgtgggctga gctgaagcat tacaatgtgc
cccggccgat gccttgcggt 2100gacttttgtg gcgtgagcat caattggcat gtctcaacgg
actacaacaa gggctggagc 2160gctcgggtga cattgttcaa ctgggaggat gtcgacatgg
ccaattggtt tgctgccatc 2220gtcatggaca aggcgtatga cggctttgag aaggcttact
cgttcaacgg caccgcagtg 2280ggcaagaaca cgatctttat gcagggtctg gaggggctta
attacctggt gaagcagacc 2340aacatgagtg ggtccgacta ccttgttcct ggcaagcaac
agtcagtcct ctcattcacc 2400aagaagctga ccccggggtt aaatgttgtt gctggagatg
gcttcccaac aaaggtcttc 2460ttcaatggcg acgaatgcgc tatgccacag agaattccga
tcagcactgg attcagcacc 2520cgtctcagca gtggccttgc tctggttccg ttccttgttg
cttcggcttt cctattgctc 2580cagcaatgat ccacgggact ccaattcttt gattctttca
ggtggtttgg tcgatgccat 2640ttgtaagaaa gcctcttttt tttgtttctg tgattgcttt
agtagattct acttagctgc 2700tgtatgttag tcagatgaag cagcagctgt gaaacagtat
gaatacttgg atagtgagag 2760aaaaagtgag gaagcatttg ttgcgggttt gcaacattgg
ttcctcgttc taatggcatt 2820tacgaattgt ctcatgttct gtctgtcata cagaatttac
tctgtgaatc cgttcatgtc 2880ttgtttttgt ttgtttggtc acaaattcag gccttgtttg
gttcctttag attgagtccg 2940tggaatggtt tctagttcga atggtttact aatatgtgta
accttaatga ggtggaatgg 3000ttcctgggtc gaatcatggt tagctgaacg gaccgtcaaa
ctgattggaa agggatcgaa 3060ggggattaaa gacggatgag gggaatttga cttgttaggg
atttagttcc ctcgttcttc 3120tccaatcccc cttagatttg agatccgaat tc
31524667PRTZea mays 4Met Ala Gly Ser Val Ala Pro
His Ala Val Val Leu Gly Leu Leu Leu1 5 10
15Leu Ala Gly Leu Ala Ala Ala Gln Arg Ala Thr Thr Pro
Ala Ala Ala 20 25 30Ala Pro
Ala Pro Asp Pro Gly Cys Asn Gly Ile Gln Leu Thr Tyr Asn 35
40 45Phe Val Asp Arg Thr Lys Ile Arg Pro Phe
Val Ser Asp Lys Asn Lys 50 55 60Gln
Pro Tyr Ala Phe Arg Ala Asn Val Thr Val Leu Asn Ser Gly Thr65
70 75 80Arg Pro Leu Lys Ser Trp
Ala Ala Leu Val Thr Phe Gly Tyr Gly Glu 85
90 95Ile Leu Val Gly Val Asp Gly Ala Val Leu Thr Gly
Gly Gly Asp Leu 100 105 110Pro
Tyr Asn Thr Thr Glu Asp Ala Gly Asn Ala Thr Ser Phe Ser Gly 115
120 125Tyr Pro His Thr Asp Leu Leu Thr Pro
Ile Ala Thr Ala Gly Asp Leu 130 135
140Ser Gln Ile Gln Ala Ser Val Gly Ile Val Gly Thr Leu Phe Ala Gly145
150 155 160Pro Gly Pro Phe
Val Pro Leu Pro Thr Ala Leu Ser Leu Asp Asp Pro 165
170 175Ala Tyr Ala Cys Pro Ala Ala Thr Asn Val
Thr Ala Arg Val Leu Ser 180 185
190Thr Cys Cys Val Leu Thr Pro Glu Ala Glu Ala Asn Ala Thr Ala Ile
195 200 205Asp Ala Asn Thr Thr Asp Pro
Thr Lys Asp Phe Leu Pro Arg Gly Thr 210 215
220Gly Asp Leu Val Ile Thr Tyr Asp Val Leu Gln Ala Tyr Pro Ser
Ser225 230 235 240Tyr Leu
Ala Leu Val Thr Leu Glu Asn Asn Ala Lys Leu Gly Arg Leu
245 250 255Asp Asn Trp Arg Leu Ser Trp
Glu Trp Arg Arg Gly Glu Phe Ile Tyr 260 265
270Ser Met Lys Gly Ala His Pro Ser Glu Val Asp Thr Ser Gly
Cys Ile 275 280 285Cys Gly Ala Pro
Gly Gln Tyr Tyr Gln Ser Leu Asp Phe Ser Gln Val 290
295 300Leu Asn Cys Asp Arg Lys Pro Val Ile Leu Asp Leu
Pro Leu Ser Arg305 310 315
320Tyr Asn Asp Thr Gln Ile Gly Lys Ile Asp Asn Cys Cys Arg Asn Gly
325 330 335Thr Ile Leu Pro Lys
Ser Met Asp Glu Ala Gln Ser Lys Ser Ala Phe 340
345 350Gln Met Gln Val Phe Lys Met Pro Pro Asp Leu Asn
Arg Thr Lys Leu 355 360 365Phe Pro
Pro Ala Asn Phe Lys Ile Val Gly Ala Ser Ser Leu Asn Pro 370
375 380Asp Tyr Ala Cys Gly Gln Pro Val Pro Val Ser
Pro Thr Ala Phe Pro385 390 395
400Asp Pro Ser Gly Leu Asp Ser Thr Thr Leu Ala Val Ala Thr Trp Gln
405 410 415Val Val Cys Asn
Ile Thr Thr Thr Lys Gly Ala Lys Pro Lys Cys Cys 420
425 430Val Thr Phe Ser Ala Tyr Tyr Asn Asp Ser Val
Ile Pro Cys Ser Thr 435 440 445Cys
Ala Cys Gly Cys Pro Ala Asn Arg Arg Gly Pro Thr Cys Ser Thr 450
455 460Thr Ala Gln Ser Met Leu Leu Pro Pro Glu
Ala Leu Leu Val Pro Phe465 470 475
480Asp Asn Arg Ser Gln Lys Ala Leu Ala Trp Ala Glu Leu Lys His
Tyr 485 490 495Asn Val Pro
Arg Pro Met Pro Cys Gly Asp Phe Cys Gly Val Ser Ile 500
505 510Asn Trp His Val Ser Thr Asp Tyr Asn Lys
Gly Trp Ser Ala Arg Val 515 520
525Thr Leu Phe Asn Trp Glu Asp Val Asp Met Ala Asn Trp Phe Ala Ala 530
535 540Ile Val Met Asp Lys Ala Tyr Asp
Gly Phe Glu Lys Ala Tyr Ser Phe545 550
555 560Asn Gly Thr Ala Val Gly Lys Asn Thr Ile Phe Met
Gln Gly Leu Glu 565 570
575Gly Leu Asn Tyr Leu Val Lys Gln Thr Asn Met Ser Gly Ser Asp Tyr
580 585 590Leu Val Pro Gly Lys Gln
Gln Ser Val Leu Ser Phe Thr Lys Lys Leu 595 600
605Thr Pro Gly Leu Asn Val Val Ala Gly Asp Gly Phe Pro Thr
Lys Val 610 615 620Phe Phe Asn Gly Asp
Glu Cys Ala Met Pro Gln Arg Ile Pro Ile Ser625 630
635 640Thr Gly Phe Ser Thr Arg Leu Ser Ser Gly
Leu Ala Leu Val Pro Phe 645 650
655Leu Val Ala Ser Ala Phe Leu Leu Leu Gln Gln 660
66552094DNAZea maysmisc_feature(2087)..(2089)n is a, c, g, or t
5ctcgtgctgc tgcttccgct gcagtaaaat acggggaaga ggaggggagg gagacgcggc
60cgctgcctgc cgcacatgct ttaagtccca ctccccacct ccccagatct ccgccctcct
120ccccaccgcc cccattcctc ccctcggccg caaccgtagc cgccgcacta cggagcaaga
180tcgtcgggta gacggacggg cgggcgggcg ggcgcggctc tgtatctatc tgtcggtggg
240agaccgcgtg tgtcggttag gcggcgggtg gcaaggaaga atggcggcga gcggcagatc
300cgtcgcgtgc tgtgccgccg cgctgctcgc ggccgcgttg ctcctctccg caccgactgc
360aacagaggct tatgattcgc tggatccaaa tggcaacatc accataaaat gggatatcat
420gcagtggact cctgatggat atgtcgctgt tgtcacaatg tttaattatc aacaatttcg
480gcatatcggc gcacctggtt ggcagcttgg gtggacatgg gcaaagaagg aggttatatg
540gtcaatggtt ggggctcaga ccactgaaca gggcgactgc tcaaagttca agagcagccc
600accccattgc tgcaagaaag atccaacaat tgtcgattta cttccaggca ctccatacaa
660catgcaaatt gccaattgct gcaaggcagg agttgtaaat acctttaacc aggacccagc
720aaatgctgct tcctccttcc agatcagtgt tggtcttgct ggaactacca ataaaactgt
780taaggtgccc aggaacttca ctcttaagac tccaggccct gggtacacat gtgggcgtgc
840cattgttggc aggcctacga agtttttcac cgcggacggg cgcagggcaa cccaagctct
900aatgacatgg aatgtgacct gcacatattc ccaatttctt gctcagaaga ctccatcctg
960ctgtgtatct ctatcatcgt tttataatga cacaattgtg aactgcccaa catgctcatg
1020tggctgccag aacccaagtg ggtcaaactg tgtgaatgag gattcaccta atctacaagc
1080tgcaattgat ggccctggca aatggactgg tcagcccctt gtacaatgca cttcccacat
1140gtgcccgata agaatccact ggcatgtgaa gctcaactac aaggattact ggagagtgaa
1200aatcactatc acaaacttca acttccgcat gaattacacg cagtggaact tagtagccca
1260gcatccaaac tttgataata tcactcagtt gttcagcttc aactacaaac cacttactcc
1320atatggtggt ggcataaatg atacggcaat gttctggggt gtaaaattct acaatgatct
1380gctgatgcaa gccggcaaac ttgggaatgt gcaatcagag ctgcttctcc gcaaggactc
1440ccggactttc actttcgaaa agggatgggc cttcccacgc cgagtttact tcaatggtga
1500taattgtgtc atgccatctc ctgaaaatta tccatggctg ccgaatgcaa gccctctaac
1560aaaaccattg gcactcccat tcttggtatt ctgggttgcc ttggctgctc tgttggctta
1620tgcatgatta gtgggatcaa gaggtttagc aagtttcaag ttgatgtcag attccatgag
1680gtgcactgca acaagtcatt tgttcattca attccatggt tgcacagaaa agatgaggcg
1740atgccaagaa aaagtcgata tgtctatgtg tttaagttaa agggccaaaa tgtatttctt
1800gtttggtata taacagccct acaacacttt ggtgaactta gttactgcag attaggtaat
1860tacagttgca ccttttgtat tttatagcaa acccagaatt tttcattgga ttctacgact
1920gcccctcttg tagtaaatgc aaggcttccc tgatactcct gtttaaagat ttgtggattg
1980ggtgagacaa tggtgattga gataactaag ttctggggtc ttgatccatt tgmwgctggr
2040aagawtattg atctaaattg ctaaaaaaaa acctcgtgcc gaattcnnng cctc
20946448PRTZea mays 6Met Ala Ala Ser Gly Arg Ser Val Ala Cys Cys Ala Ala
Ala Leu Leu1 5 10 15Ala
Ala Ala Leu Leu Leu Ser Ala Pro Thr Ala Thr Glu Ala Tyr Asp 20
25 30Ser Leu Asp Pro Asn Gly Asn Ile
Thr Ile Lys Trp Asp Ile Met Gln 35 40
45Trp Thr Pro Asp Gly Tyr Val Ala Val Val Thr Met Phe Asn Tyr Gln
50 55 60Gln Phe Arg His Ile Gly Ala Pro
Gly Trp Gln Leu Gly Trp Thr Trp65 70 75
80Ala Lys Lys Glu Val Ile Trp Ser Met Val Gly Ala Gln
Thr Thr Glu 85 90 95Gln
Gly Asp Cys Ser Lys Phe Lys Ser Ser Pro Pro His Cys Cys Lys
100 105 110Lys Asp Pro Thr Ile Val Asp
Leu Leu Pro Gly Thr Pro Tyr Asn Met 115 120
125Gln Ile Ala Asn Cys Cys Lys Ala Gly Val Val Asn Thr Phe Asn
Gln 130 135 140Asp Pro Ala Asn Ala Ala
Ser Ser Phe Gln Ile Ser Val Gly Leu Ala145 150
155 160Gly Thr Thr Asn Lys Thr Val Lys Val Pro Arg
Asn Phe Thr Leu Lys 165 170
175Thr Pro Gly Pro Gly Tyr Thr Cys Gly Arg Ala Ile Val Gly Arg Pro
180 185 190Thr Lys Phe Phe Thr Ala
Asp Gly Arg Arg Ala Thr Gln Ala Leu Met 195 200
205Thr Trp Asn Val Thr Cys Thr Tyr Ser Gln Phe Leu Ala Gln
Lys Thr 210 215 220Pro Ser Cys Cys Val
Ser Leu Ser Ser Phe Tyr Asn Asp Thr Ile Val225 230
235 240Asn Cys Pro Thr Cys Ser Cys Gly Cys Gln
Asn Pro Ser Gly Ser Asn 245 250
255Cys Val Asn Glu Asp Ser Pro Asn Leu Gln Ala Ala Ile Asp Gly Pro
260 265 270Gly Lys Trp Thr Gly
Gln Pro Leu Val Gln Cys Thr Ser His Met Cys 275
280 285Pro Ile Arg Ile His Trp His Val Lys Leu Asn Tyr
Lys Asp Tyr Trp 290 295 300Arg Val Lys
Ile Thr Ile Thr Asn Phe Asn Phe Arg Met Asn Tyr Thr305
310 315 320Gln Trp Asn Leu Val Ala Gln
His Pro Asn Phe Asp Asn Ile Thr Gln 325
330 335Leu Phe Ser Phe Asn Tyr Lys Pro Leu Thr Pro Tyr
Gly Gly Gly Ile 340 345 350Asn
Asp Thr Ala Met Phe Trp Gly Val Lys Phe Tyr Asn Asp Leu Leu 355
360 365Met Gln Ala Gly Lys Leu Gly Asn Val
Gln Ser Glu Leu Leu Leu Arg 370 375
380Lys Asp Ser Arg Thr Phe Thr Phe Glu Lys Gly Trp Ala Phe Pro Arg385
390 395 400Arg Val Tyr Phe
Asn Gly Asp Asn Cys Val Met Pro Ser Pro Glu Asn 405
410 415Tyr Pro Trp Leu Pro Asn Ala Ser Pro Leu
Thr Lys Pro Leu Ala Leu 420 425
430Pro Phe Leu Val Phe Trp Val Ala Leu Ala Ala Leu Leu Ala Tyr Ala
435 440 44572102DNAZea mays 7ggaaagcagc
gctgcggagc agagtgtgtc gcttcgctgt aaaaacaggg gagagggaga 60cgcgcccgct
gccagtgcct gccgcacacg cgtttagcgt ttaagttcca ctcctcgccg 120ccccagatct
ccgccctcct caccactgcc cctcattccc cggcgcccag cacccggcgg 180ccgcaaccgc
cgcagtccgg agcaagatcg gcgggtagac ggacggacgg acgggcgaca 240ggcgggcggg
cgcggctctg tctgtatcta tctgttggtg ggagaccggt tgtgtcggtt 300aggcggcggc
gggtgggaag gaagaatggc ggcgggcggc agatccatcg cgtgctttgc 360cgccgtgctg
ctcgcggccg cgctgctcct ctccgcaccg accaccacag aggcctacga 420ttcgctggat
ccaaacggca acatcactat aaaatgggat atcatgcagt ggactcctga 480cggatatgtc
gctgttgtca caatgttcaa ttatcaacaa tttcggcaca tcggggcacc 540tggatggcag
cttgggtgga catgggcaaa aaaggaggtt atatggtcaa tggttggggc 600tcagaccact
gaacagggtg actgctcaaa gttcaagggc aacacccccc attgctgcaa 660gaaagatcca
acaattgttg atttacttcc aggcactcca tacaacatgc aaattgccaa 720ttgctgcaag
gcaggagtta taaatacctt taaccaggac ccagcaaatg ctgcttcctc 780cttccagatc
agtgttggtc ttgctggaac taccaataaa actgttaagg tgccgaagaa 840tttcactctt
aagactccag gccctgggta cacatgtggg cgtgctattg ttggcaggcc 900aacgaagttt
ttctctgcag atgggcgcag ggtaacccaa gctctaatga catggaatgt 960gacctgcaca
tattcccaat ttcttgctca gaagactcca tcctgctgtg tatctctctc 1020atcattttat
aatgacacaa ttgtgaactg cccgacatgc tcatgtggct gccagaaccc 1080aagtgggtca
aactgtgtga acgaggattc acctaatcta caagccgcaa ttgatggtcc 1140tggtaaatgg
actggccagc ctcttgtaca atgcacttct cacatgtgcc caataagaat 1200ccactggcat
gtgaagctca actacaagga atactggaga gtgaaaatca ctatcacgaa 1260cttcaacttc
cgcatgaatt acacacagtg gaacttagtt gctcagcatc caaactttga 1320taatatcact
cagttgttca gcttcaacta caaaccactt actccatatg ggggtggcat 1380aaatgatacg
gcaatgttct ggggtgtaaa gttctacaat gatttgctga tgcaagccgg 1440caaacttggg
aatgtgcaat cagaactgct tctccgcaag gactcacgga ctttcacatt 1500cgaaaaggga
tgggccttcc cacgccgagt gtacttcaat ggtgataatt gtgtcatgcc 1560atctcctgaa
aattatccat ggctgccgaa tgcaagccct ctaacaaaac aagcattgac 1620actcccactc
ttgatattct gggttgcctt ggctgttctg ttggcttatg catgatgagt 1680gggatcaaga
tgtttagcaa gcttcaagtt gatgtcggat tccatgaggt gcactgcaac 1740gggatattta
ttcattcaat tccatagcgg cacaggagag atgaggcgaa gccaagaaaa 1800agtggatgtg
tgtgtgtgtg tgtttgtaag ttaaagggcc aaaatgtatt tcttgtctgg 1860tagtatatag
cagctctaca acactttggt gaacttagtt actgcaaatt aggcaattac 1920agttgcacct
tttgtatttt atagcaaacc cagacttcta ttggattcta tgactgcccc 1980tcttgtagta
aacgcaaggc ttcactggta ctcctgttta aagattggtc aaatagaaga 2040gacgacggtg
attgtcaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa 2100aa
21028449PRTZea
mays 8Met Ala Ala Gly Gly Arg Ser Ile Ala Cys Phe Ala Ala Val Leu Leu1
5 10 15Ala Ala Ala Leu Leu
Leu Ser Ala Pro Thr Thr Thr Glu Ala Tyr Asp 20
25 30Ser Leu Asp Pro Asn Gly Asn Ile Thr Ile Lys Trp
Asp Ile Met Gln 35 40 45Trp Thr
Pro Asp Gly Tyr Val Ala Val Val Thr Met Phe Asn Tyr Gln 50
55 60Gln Phe Arg His Ile Gly Ala Pro Gly Trp Gln
Leu Gly Trp Thr Trp65 70 75
80Ala Lys Lys Glu Val Ile Trp Ser Met Val Gly Ala Gln Thr Thr Glu
85 90 95Gln Gly Asp Cys Ser
Lys Phe Lys Gly Asn Thr Pro His Cys Cys Lys 100
105 110Lys Asp Pro Thr Ile Val Asp Leu Leu Pro Gly Thr
Pro Tyr Asn Met 115 120 125Gln Ile
Ala Asn Cys Cys Lys Ala Gly Val Ile Asn Thr Phe Asn Gln 130
135 140Asp Pro Ala Asn Ala Ala Ser Ser Phe Gln Ile
Ser Val Gly Leu Ala145 150 155
160Gly Thr Thr Asn Lys Thr Val Lys Val Pro Lys Asn Phe Thr Leu Lys
165 170 175Thr Pro Gly Pro
Gly Tyr Thr Cys Gly Arg Ala Ile Val Gly Arg Pro 180
185 190Thr Lys Phe Phe Ser Ala Asp Gly Arg Arg Val
Thr Gln Ala Leu Met 195 200 205Thr
Trp Asn Val Thr Cys Thr Tyr Ser Gln Phe Leu Ala Gln Lys Thr 210
215 220Pro Ser Cys Cys Val Ser Leu Ser Ser Phe
Tyr Asn Asp Thr Ile Val225 230 235
240Asn Cys Pro Thr Cys Ser Cys Gly Cys Gln Asn Pro Ser Gly Ser
Asn 245 250 255Cys Val Asn
Glu Asp Ser Pro Asn Leu Gln Ala Ala Ile Asp Gly Pro 260
265 270Gly Lys Trp Thr Gly Gln Pro Leu Val Gln
Cys Thr Ser His Met Cys 275 280
285Pro Ile Arg Ile His Trp His Val Lys Leu Asn Tyr Lys Glu Tyr Trp 290
295 300Arg Val Lys Ile Thr Ile Thr Asn
Phe Asn Phe Arg Met Asn Tyr Thr305 310
315 320Gln Trp Asn Leu Val Ala Gln His Pro Asn Phe Asp
Asn Ile Thr Gln 325 330
335Leu Phe Ser Phe Asn Tyr Lys Pro Leu Thr Pro Tyr Gly Gly Gly Ile
340 345 350Asn Asp Thr Ala Met Phe
Trp Gly Val Lys Phe Tyr Asn Asp Leu Leu 355 360
365Met Gln Ala Gly Lys Leu Gly Asn Val Gln Ser Glu Leu Leu
Leu Arg 370 375 380Lys Asp Ser Arg Thr
Phe Thr Phe Glu Lys Gly Trp Ala Phe Pro Arg385 390
395 400Arg Val Tyr Phe Asn Gly Asp Asn Cys Val
Met Pro Ser Pro Glu Asn 405 410
415Tyr Pro Trp Leu Pro Asn Ala Ser Pro Leu Thr Lys Gln Ala Leu Thr
420 425 430Leu Pro Leu Leu Ile
Phe Trp Val Ala Leu Ala Val Leu Leu Ala Tyr 435
440 445Ala92422DNAZea mays 9aaccatgcac ctcaatttta
gcaacctcgc acaaaaactg catgcctaat tataccttct 60tggcccctcc tcccccgctg
gaacgcatgt gtttgcttcc catcgctcct ggcttccccg 120tcgagcgagg gagccacatt
cttgcttcca ttgttcgctc aaacgcttgg tagctcgatc 180ggcgccgttg ttcttgggcc
ggccggtcga gccgaatggt catggcagcg ccagtgccgc 240tccggcggcg gcgggcgctc
ctggtggtag cgacgttact cgccgtggtc accgcggcga 300tggcgcagga ctataaagat
ggtggcggcg acgactacga ggaggacgag aagaagaagc 360cgcagttcaa ggcgcaggag
gcgtgcaacg gcgtgttcct gacgtacacg ttcatggagc 420gcgccaagga gtacccgcac
ctgaagaagg cggcggcgca gccgtacgcg ttcaaggcca 480cggcgacggt gctcaacacc
atgaccgagg acctcaaggc gtggcagatg ttcgtgggct 540tccagcacaa ggagatcctc
gtgtccgtcg gcggcgccgt gctgctcgac ggctccgacc 600tccccgccaa cgtgtccggt
ggcgccacct ttgcgggata cccaatggcc gacctcctca 660actccatcga gacggcgggc
gagccgtccc tgatcgagag caagattgag atcaccggca 720cccaattcgg cgtgaaggcc
cccgggaagc ccatgcccaa gaccatcaag ttgaccaacc 780ccgtgggctt ccggtgcccc
gcccccaacc acaaagacag cgtgatgtac gtgtgctgcg 840tcaaggaccg caagttcaag
gcgaagaagg ctaacagcac gcggtaccag acacggcgga 900aagcggacct gacgttcgcc
tacgacgtgc tgcaggccaa caccaacaac taccaggtgc 960aggtgaccat cgacaactgg
agccccatca gccggctgga caactggaac ctcacctggg 1020agtggaagcg cggcgagttc
atctacagca tgaagggcgc ctacacgctg ctcaaggaag 1080gccccgcctg catctacagc
cccgcagcgg gctactacaa ggacatggac ttcacccccg 1140tctacaactg cgagaagcgg
cccgtcatcg tggacctccc gccggagcgg gagaaggacg 1200acgccgtcgg gaacctcccc
ttctgctgca agaacggcac gctgctgccg cccaccatgg 1260acccgtccaa gtcgcgggcc
atgttccaga tgcaggtgta caagctgccg ccggacctga 1320accgcacggc gctgtacccg
ccgcagaact ggaagatctc cggcaagctc aacccgcagt 1380acgcgtgcgg gccgcccgtc
cgcgtgagcc cccaggagtt cccggacccg acgggtctca 1440tgtcgaccac ccccgccgtg
gcgtcgtggc aggtggcgtg caacatcacg cggcccaaga 1500agcgcgcctc caagtgctgc
gtctccttct ccgcctacta caacgactcc gtggtgccgt 1560gcaacacctg cgcctgcggc
tgcggcgacg acaccgcgac gtgcgacccg gacaagcgcg 1620ccatgctgct gccaccggag
gcgctgctcg tcccgttcga caaccggtcg gccaaggcac 1680gggcgtgggc caagatcaag
cactggcggg tgcccaaccc catgccgtgc agcgacaact 1740gcggcgtcag catcaactgg
cacgtcatca acaactacaa gtccggctgg tcggcgcgca 1800tgaccatctt caactggcag
gactacacct tcaaggattg gtttgccgca gtgaccatgg 1860gcagccactt cagcggctac
gagaacgtct actccttcaa cggcacgcgg atgggcgccc 1920ccttcaacaa caccatcttc
atgcaggggg tgccgggcct cgcttacctc gagcccatca 1980ccgacgcgaa gacgacatcg
gaacccaggc ttcccggcaa gcagcagtcg gtcatctcgt 2040tcaccaggaa agacgcgccc
aatgtcaaca ttcccagagg ggaaggcttc cccaagagga 2100tctacttcga cggcgaggag
tgcgcgctcc cggataggat acccaaggtg tcgagcgcgc 2160gccggcgggc tgggaccgcg
agcctgggtc agatagccat ggcggcggcg ctcgtgatga 2220ttgtggcgct agtggattcc
ttgtgcctat gatgactgaa aacttctttg gttcatagag 2280gatttgactg acctagcgcg
ctgcatttgt tgaacacttc attcattaac taggtacgtg 2340cccgtgcgtt gctacggaag
taaaaaaata gcgtacaaaa tatatacgaa gcgaaaaaca 2400tcactatgat agtaaaaatc
gt 242210678PRTZea mays 10Met
Val Met Ala Ala Pro Val Pro Leu Arg Arg Arg Arg Ala Leu Leu1
5 10 15Val Val Ala Thr Leu Leu Ala
Val Val Thr Ala Ala Met Ala Gln Asp 20 25
30Tyr Lys Asp Gly Gly Gly Asp Asp Tyr Glu Glu Asp Glu Lys
Lys Lys 35 40 45Pro Gln Phe Lys
Ala Gln Glu Ala Cys Asn Gly Val Phe Leu Thr Tyr 50 55
60Thr Phe Met Glu Arg Ala Lys Glu Tyr Pro His Leu Lys
Lys Ala Ala65 70 75
80Ala Gln Pro Tyr Ala Phe Lys Ala Thr Ala Thr Val Leu Asn Thr Met
85 90 95Thr Glu Asp Leu Lys Ala
Trp Gln Met Phe Val Gly Phe Gln His Lys 100
105 110Glu Ile Leu Val Ser Val Gly Gly Ala Val Leu Leu
Asp Gly Ser Asp 115 120 125Leu Pro
Ala Asn Val Ser Gly Gly Ala Thr Phe Ala Gly Tyr Pro Met 130
135 140Ala Asp Leu Leu Asn Ser Ile Glu Thr Ala Gly
Glu Pro Ser Leu Ile145 150 155
160Glu Ser Lys Ile Glu Ile Thr Gly Thr Gln Phe Gly Val Lys Ala Pro
165 170 175Gly Lys Pro Met
Pro Lys Thr Ile Lys Leu Thr Asn Pro Val Gly Phe 180
185 190Arg Cys Pro Ala Pro Asn His Lys Asp Ser Val
Met Tyr Val Cys Cys 195 200 205Val
Lys Asp Arg Lys Phe Lys Ala Lys Lys Ala Asn Ser Thr Arg Tyr 210
215 220Gln Thr Arg Arg Lys Ala Asp Leu Thr Phe
Ala Tyr Asp Val Leu Gln225 230 235
240Ala Asn Thr Asn Asn Tyr Gln Val Gln Val Thr Ile Asp Asn Trp
Ser 245 250 255Pro Ile Ser
Arg Leu Asp Asn Trp Asn Leu Thr Trp Glu Trp Lys Arg 260
265 270Gly Glu Phe Ile Tyr Ser Met Lys Gly Ala
Tyr Thr Leu Leu Lys Glu 275 280
285Gly Pro Ala Cys Ile Tyr Ser Pro Ala Ala Gly Tyr Tyr Lys Asp Met 290
295 300Asp Phe Thr Pro Val Tyr Asn Cys
Glu Lys Arg Pro Val Ile Val Asp305 310
315 320Leu Pro Pro Glu Arg Glu Lys Asp Asp Ala Val Gly
Asn Leu Pro Phe 325 330
335Cys Cys Lys Asn Gly Thr Leu Leu Pro Pro Thr Met Asp Pro Ser Lys
340 345 350Ser Arg Ala Met Phe Gln
Met Gln Val Tyr Lys Leu Pro Pro Asp Leu 355 360
365Asn Arg Thr Ala Leu Tyr Pro Pro Gln Asn Trp Lys Ile Ser
Gly Lys 370 375 380Leu Asn Pro Gln Tyr
Ala Cys Gly Pro Pro Val Arg Val Ser Pro Gln385 390
395 400Glu Phe Pro Asp Pro Thr Gly Leu Met Ser
Thr Thr Pro Ala Val Ala 405 410
415Ser Trp Gln Val Ala Cys Asn Ile Thr Arg Pro Lys Lys Arg Ala Ser
420 425 430Lys Cys Cys Val Ser
Phe Ser Ala Tyr Tyr Asn Asp Ser Val Val Pro 435
440 445Cys Asn Thr Cys Ala Cys Gly Cys Gly Asp Asp Thr
Ala Thr Cys Asp 450 455 460Pro Asp Lys
Arg Ala Met Leu Leu Pro Pro Glu Ala Leu Leu Val Pro465
470 475 480Phe Asp Asn Arg Ser Ala Lys
Ala Arg Ala Trp Ala Lys Ile Lys His 485
490 495Trp Arg Val Pro Asn Pro Met Pro Cys Ser Asp Asn
Cys Gly Val Ser 500 505 510Ile
Asn Trp His Val Ile Asn Asn Tyr Lys Ser Gly Trp Ser Ala Arg 515
520 525Met Thr Ile Phe Asn Trp Gln Asp Tyr
Thr Phe Lys Asp Trp Phe Ala 530 535
540Ala Val Thr Met Gly Ser His Phe Ser Gly Tyr Glu Asn Val Tyr Ser545
550 555 560Phe Asn Gly Thr
Arg Met Gly Ala Pro Phe Asn Asn Thr Ile Phe Met 565
570 575Gln Gly Val Pro Gly Leu Ala Tyr Leu Glu
Pro Ile Thr Asp Ala Lys 580 585
590Thr Thr Ser Glu Pro Arg Leu Pro Gly Lys Gln Gln Ser Val Ile Ser
595 600 605Phe Thr Arg Lys Asp Ala Pro
Asn Val Asn Ile Pro Arg Gly Glu Gly 610 615
620Phe Pro Lys Arg Ile Tyr Phe Asp Gly Glu Glu Cys Ala Leu Pro
Asp625 630 635 640Arg Ile
Pro Lys Val Ser Ser Ala Arg Arg Arg Ala Gly Thr Ala Ser
645 650 655Leu Gly Gln Ile Ala Met Ala
Ala Ala Leu Val Met Ile Val Ala Leu 660 665
670Val Asp Ser Leu Cys Leu 675111845DNAZea mays
11agaagagaga ggggaaagtt gcttctctct ctgacccgcc cactccctcc ttccctgctc
60cggtcgcact ccgtagttcc tccgcgcact tacgtacagc agacagacac gagatcgagt
120ggtacagggc ccgccagaaa cctcacgagc tagctgggtt cctgccgcgc cgccgatcca
180cgcatggcgc cgccgccgct cctgcccgcg cgcttcgtcg ccgcctccgt cgcgctgctc
240gccgtcgcct tctcctcctc tctaacgcgt ccgtcaggtg catacgatcc gctcgatccg
300aacgggaaca taacaatcaa gtgggacgtg atacagtgga ctgcggatgg ctatgtggcc
360gtcgtttcgc tatacaacta ccagcagtac cgccacatcc aggcgccgcc ggggtggagg
420ctaggctggg tgtgggcgaa gaaggaggtg atctgggcga tgaccggcgg ccaggccacc
480gagcagggcg actgctccag gttcaaggcc agcgtcctcc cccactgctg caggagggac
540ccggaggtgg tggacctgct gcccgggact ccctacaaca cgcagaccgc caactgctgc
600aggggaggag tgctcgcctc gtgggcgcag gaccctagcg acgccgtcgc ctcgttccag
660gtcagcgttg ggcaggctgg gtccaccaac aggaccgtca aggtgcccag gaacttcacc
720ctgctggcgc ctggtcccgg gtacacctgc ggagccgcca agcttgtcaa gcctaccaag
780ttcatgtctc aggatggcag gagatcaact caagcgcaca tgacctggaa cgtgacgtgc
840acgtactccc agttccttgc ccagagatct ccaacctgct gtgtctcgct ctcgtcgttc
900tacaacgaca ccattgttag ctgcccagca tgctcctgcg gctgccagaa caacaacagc
960agtagcaccg ccgcgccagg aagctgcgta gagggtagta gaaggtcgcc ctatctggct
1020tccgtcgtca acgatcctag caagaacagc ttggcgccgc tagtccagtg cacctcacac
1080atgtgcccgg taagggtgca ctggcacgtc aaggtcagct acaaggagta ctggagggtg
1140aagatcacgg tcaccaactt caactaccgg atgaactact cgcagtggaa cctggtcgcg
1200cagcacccca acttcgacaa cctcaccacc attttcagct tcaactacag acctctcaac
1260ccctacggag tgatcaacga cacggcgatg ctatggggca tcaagtacta caacgatctg
1320ctcatgacgg ccgggccaga cgggaacgtg cagtccgagc ttctgttccg gaaggagccg
1380tccacgttca ccttccacaa aggatgggcc ttccccaggc gagtctactt caacggagac
1440aactgcgtga tgccgccgcc ggacgcctac ccgtggctgc ccaacgccgc ctcgccgcgg
1500ctgtcgcctt cgcttctcct cccgctcgtt gcggctgctt ggacagcatt cgcagtcctt
1560tcgtgatggg cccatatgcg tagggaaggc aaggcaaggc acacaatgtc ccatgacaag
1620ttctgacctg attcagcgtt gttgcttgct gctgatcatt agtcgatctg ttgcgaagtt
1680ttatttggtg tcttgaatct tgattcagga acaggttcag atgtgcattc acgtactacc
1740aagcatgtac attcccaata cttgtaaatt tctgcaaaga ctgactggca agtgacagta
1800gaaataatct gtttctctct ccgcatcagg aaagtttcgg ctcaa
184512460PRTZea mays 12Met Ala Pro Pro Pro Leu Leu Pro Ala Arg Phe Val
Ala Ala Ser Val1 5 10
15Ala Leu Leu Ala Val Ala Phe Ser Ser Ser Leu Thr Arg Pro Ser Gly
20 25 30Ala Tyr Asp Pro Leu Asp Pro
Asn Gly Asn Ile Thr Ile Lys Trp Asp 35 40
45Val Ile Gln Trp Thr Ala Asp Gly Tyr Val Ala Val Val Ser Leu
Tyr 50 55 60Asn Tyr Gln Gln Tyr Arg
His Ile Gln Ala Pro Pro Gly Trp Arg Leu65 70
75 80Gly Trp Val Trp Ala Lys Lys Glu Val Ile Trp
Ala Met Thr Gly Gly 85 90
95Gln Ala Thr Glu Gln Gly Asp Cys Ser Arg Phe Lys Ala Ser Val Leu
100 105 110Pro His Cys Cys Arg Arg
Asp Pro Glu Val Val Asp Leu Leu Pro Gly 115 120
125Thr Pro Tyr Asn Thr Gln Thr Ala Asn Cys Cys Arg Gly Gly
Val Leu 130 135 140Ala Ser Trp Ala Gln
Asp Pro Ser Asp Ala Val Ala Ser Phe Gln Val145 150
155 160Ser Val Gly Gln Ala Gly Ser Thr Asn Arg
Thr Val Lys Val Pro Arg 165 170
175Asn Phe Thr Leu Leu Ala Pro Gly Pro Gly Tyr Thr Cys Gly Ala Ala
180 185 190Lys Leu Val Lys Pro
Thr Lys Phe Met Ser Gln Asp Gly Arg Arg Ser 195
200 205Thr Gln Ala His Met Thr Trp Asn Val Thr Cys Thr
Tyr Ser Gln Phe 210 215 220Leu Ala Gln
Arg Ser Pro Thr Cys Cys Val Ser Leu Ser Ser Phe Tyr225
230 235 240Asn Asp Thr Ile Val Ser Cys
Pro Ala Cys Ser Cys Gly Cys Gln Asn 245
250 255Asn Asn Ser Ser Ser Thr Ala Ala Pro Gly Ser Cys
Val Glu Gly Ser 260 265 270Arg
Arg Ser Pro Tyr Leu Ala Ser Val Val Asn Asp Pro Ser Lys Asn 275
280 285Ser Leu Ala Pro Leu Val Gln Cys Thr
Ser His Met Cys Pro Val Arg 290 295
300Val His Trp His Val Lys Val Ser Tyr Lys Glu Tyr Trp Arg Val Lys305
310 315 320Ile Thr Val Thr
Asn Phe Asn Tyr Arg Met Asn Tyr Ser Gln Trp Asn 325
330 335Leu Val Ala Gln His Pro Asn Phe Asp Asn
Leu Thr Thr Ile Phe Ser 340 345
350Phe Asn Tyr Arg Pro Leu Asn Pro Tyr Gly Val Ile Asn Asp Thr Ala
355 360 365Met Leu Trp Gly Ile Lys Tyr
Tyr Asn Asp Leu Leu Met Thr Ala Gly 370 375
380Pro Asp Gly Asn Val Gln Ser Glu Leu Leu Phe Arg Lys Glu Pro
Ser385 390 395 400Thr Phe
Thr Phe His Lys Gly Trp Ala Phe Pro Arg Arg Val Tyr Phe
405 410 415Asn Gly Asp Asn Cys Val Met
Pro Pro Pro Asp Ala Tyr Pro Trp Leu 420 425
430Pro Asn Ala Ala Ser Pro Arg Leu Ser Pro Ser Leu Leu Leu
Pro Leu 435 440 445Val Ala Ala Ala
Trp Thr Ala Phe Ala Val Leu Ser 450 455
460131644DNAZea mays 13tgcacgcccg atactgctag ccaaggccaa gccagtgcag
gcgcggtggt gtgtgttgtt 60ctcgtcgcgc actcgccggc agcgatggag ccccgccgct
ccgtgctgct cctggccctc 120gccgtcgccg ccgcgctctc cgtcgcagtg gcttacgacc
cgttggaccc gaacgggaac 180attaccatca agtgggacat catgtcgtgg acgcccgacg
gctatgtcgc ggtggtgacc 240atcaacaact tccagacgta ccggcagatc acggcgccgg
ggtggacggt ggggtggacg 300tgggcgaagc gggaggtgat ctggtccatg gtgggcgcgc
aggccacgga gcagggcgac 360tgctcccgct tcaaggccaa catcccgcac tgctgcaagc
gcaccccggc cgtcgtcgac 420ctgctccccg gcgtgcccta caaccagcag atcgccaact
gctgccgcgg cggcgtcgtc 480agcgcctacg gccaggaccc ggccaccgcc gtcgccgcgt
tccaggtcag cgtcggccag 540gccggcacca ccaaccgcac cgtcaaggtg cccaagaact
tcacgctgct ggggccgggg 600ccaggataca cctgcggccc cggcaaggtc gtcccctcca
ccgtcttcct cacgcccgac 660cgccgacgca agacacaagc cctcatgacg tggaacgtga
cgtgcaccta ctcgcagcac 720ctggcgtcca agtacccctc ctgctgcgtc tccttctcct
ccttctacaa cgacaccatc 780gtgccctgcg ccaagtgcgc ctgcggctgc gagcacaaga
cctgcgtcca gggcgactcg 840aagcggctgg cggtgacggg gaagcacgcg cacacggcgg
cggcggtgcg cgggcagcac 900cgggacaagg aggcgccgct gctgcagtgc acgacgcaca
tgtgccccgt gcgcgtgcac 960tggcacgtca agctcaacta caaggagtac tggcgcgcca
agatcgccat caccaacttc 1020aactaccaca tgaactacac gcagtggacg ctcgtcgcgc
agcaccccaa cctcgacaac 1080atcaccgagg tcttcagctt cggctacaag cccgtcgtct
cctatggatc catcaatgac 1140acggccatgt tctacgggct caagtacttc aacgaccacc
tgatgcaggc ggggccgtac 1200gggaacgtgc agtcggaggt gctcatgcgc aaggacgcca
gcaccttcac cttcaggcag 1260ggctgggcct tcccgcgcaa ggtctacttc aacggcgacg
agtgccagat gccgccgccg 1320gacgcctacc cctacttgcc caactccgcg ccgccgacag
ccgcggcgtc gctgggcggc 1380gcagcggcag cggccgtcgt ggtgctcttg ggcatgatcg
tggcatgaga aaacacggga 1440catcgatcga cctagtgcta ggaccggcac aggggaatgg
aaaaaagacg ttgctttctt 1500ctgtagatag agagaccaga gacctcggtt tgggtttcag
gaatggtttg gaactttgga 1560tgtttttctt tcagtgtaga tggacaagcc atgattttgc
aaggaaaatt aacatgtgca 1620aaaaaaaaaa aaaaaaaaaa aaaa
164414447PRTZea mays 14Met Glu Pro Arg Arg Ser Val
Leu Leu Leu Ala Leu Ala Val Ala Ala1 5 10
15Ala Leu Ser Val Ala Val Ala Tyr Asp Pro Leu Asp Pro
Asn Gly Asn 20 25 30Ile Thr
Ile Lys Trp Asp Ile Met Ser Trp Thr Pro Asp Gly Tyr Val 35
40 45Ala Val Val Thr Ile Asn Asn Phe Gln Thr
Tyr Arg Gln Ile Thr Ala 50 55 60Pro
Gly Trp Thr Val Gly Trp Thr Trp Ala Lys Arg Glu Val Ile Trp65
70 75 80Ser Met Val Gly Ala Gln
Ala Thr Glu Gln Gly Asp Cys Ser Arg Phe 85
90 95Lys Ala Asn Ile Pro His Cys Cys Lys Arg Thr Pro
Ala Val Val Asp 100 105 110Leu
Leu Pro Gly Val Pro Tyr Asn Gln Gln Ile Ala Asn Cys Cys Arg 115
120 125Gly Gly Val Val Ser Ala Tyr Gly Gln
Asp Pro Ala Thr Ala Val Ala 130 135
140Ala Phe Gln Val Ser Val Gly Gln Ala Gly Thr Thr Asn Arg Thr Val145
150 155 160Lys Val Pro Lys
Asn Phe Thr Leu Leu Gly Pro Gly Pro Gly Tyr Thr 165
170 175Cys Gly Pro Gly Lys Val Val Pro Ser Thr
Val Phe Leu Thr Pro Asp 180 185
190Arg Arg Arg Lys Thr Gln Ala Leu Met Thr Trp Asn Val Thr Cys Thr
195 200 205Tyr Ser Gln His Leu Ala Ser
Lys Tyr Pro Ser Cys Cys Val Ser Phe 210 215
220Ser Ser Phe Tyr Asn Asp Thr Ile Val Pro Cys Ala Lys Cys Ala
Cys225 230 235 240Gly Cys
Glu His Lys Thr Cys Val Gln Gly Asp Ser Lys Arg Leu Ala
245 250 255Val Thr Gly Lys His Ala His
Thr Ala Ala Ala Val Arg Gly Gln His 260 265
270Arg Asp Lys Glu Ala Pro Leu Leu Gln Cys Thr Thr His Met
Cys Pro 275 280 285Val Arg Val His
Trp His Val Lys Leu Asn Tyr Lys Glu Tyr Trp Arg 290
295 300Ala Lys Ile Ala Ile Thr Asn Phe Asn Tyr His Met
Asn Tyr Thr Gln305 310 315
320Trp Thr Leu Val Ala Gln His Pro Asn Leu Asp Asn Ile Thr Glu Val
325 330 335Phe Ser Phe Gly Tyr
Lys Pro Val Val Ser Tyr Gly Ser Ile Asn Asp 340
345 350Thr Ala Met Phe Tyr Gly Leu Lys Tyr Phe Asn Asp
His Leu Met Gln 355 360 365Ala Gly
Pro Tyr Gly Asn Val Gln Ser Glu Val Leu Met Arg Lys Asp 370
375 380Ala Ser Thr Phe Thr Phe Arg Gln Gly Trp Ala
Phe Pro Arg Lys Val385 390 395
400Tyr Phe Asn Gly Asp Glu Cys Gln Met Pro Pro Pro Asp Ala Tyr Pro
405 410 415Tyr Leu Pro Asn
Ser Ala Pro Pro Thr Ala Ala Ala Ser Leu Gly Gly 420
425 430Ala Ala Ala Ala Ala Val Val Val Leu Leu Gly
Met Ile Val Ala 435 440
445152108DNAZea mays 15tcttcgtctt cgtgtcatcg tgcgtgtgtg cgcttggacg
gaaacggcct ctcatcctcc 60gacgacccag actcgatcca ctcaccacca ccggcttatt
cgtttatacg gaaacagatt 120cagcttctgt gcctgcttca gccatggcca tgctgcccgt
cgtcgtccgc atggccgcca 180tgcccttcat cttccttgtg ctcctcgcgc acaccgcctc
ggcacagccc gacgccggct 240gcaacgggat cctcctcacc tacacgctgc agcgccggga
caagatcagg cctcacgtgg 300cggcgcccaa ctcccagccc tactccttca gcgccagcgc
caccgtcgtc aacgccggca 360cccgcccgct acgctcctgg gcgctgctgc tcaccttcgt
gcacggcgag atcctcgtct 420ccgtcgacgg ggccgtgctc acctcgggcg ccgccctgcc
ctacaacacc acggcggggg 480acgccgccgg caggcccacg cccacgtcct tcaccgggta
cccgcagacg gacctcctca 540ccccgatcgc cacggccggg gaccccgcca agacacaggc
cacggtcagc ctcgtaggca 600cgctcttcgc cgggccggag ccctacgtcc cgctcccctc
gtttctctcg ctcgccgacc 660cttcctacac ctgcccgccg gccaccaacg ccacgtcgtc
gccgacgaac ctcaccacct 720gctgcgtgtt cacggcgggt ggggacccca ccggcggcct
ggtggagagt ggcttcctcc 780cgcgccgcac cggcgacctg gtcatcacct acgacgtgct
ccagtcgtac gacaccacct 840acctagcgct cgtcacgctg gagaacgacg cgctgctcgg
ccgcctcgac gcctggcagc 900tgtcgtggag gtgggagcac ggggagttca tcagctccat
gcgaggcgcc tacccgcggg 960aggtggacac ggccgaatgc ctctacggtc cccagggcca
gtactacaag gacctcgact 1020tctccaaggc ggtgctcaac tgcgaccgca ggcccgtcgt
ccacgacctg ccgccgtcgc 1080gggccaacga cacggagatc ggccggatcg accactgctg
ccggaacggc accatcctgc 1140ccaagtccat ggacgtcgcg cgctccaagt cggcgttcca
gatggtggtg tacaagatgc 1200cgcccgacct caaccggacc aagctctacc cgcccacagg
gttcaacgtc accggcgccg 1260cgtccgcgct gaacccggag tacgcgtgcg acccacccat
ctcggtgagc ccgtcggagt 1320acccggaccc cagcgggctc acgtcgatca cggtggccgt
ggcgacgtgg caggtggtgt 1380gcaacatcac cacgtcgccc aagaagccgc ccaggtgctg
cgtctccttc tcctccttct 1440acaacgagtc ggtggtcccc tgccggacgt gcgcgtgcgg
ctgcccctcg tccgcgccga 1500cctgcagcac cacggcgccg gcgatgctgc tgccgccgca
ggcgctgctc atgccgttcg 1560accggcgggc cagcgaggcg ctcgagtggg cggaccagaa
gcacctcggc gtgcccaaac 1620ccatgccctg cggcgacttc tgcggcgtca gcgtcaactg
gcacgtcgcc accgacttca 1680ccggaggatg gagcgcgcgc ctcacgctct tcaactggga
cggcacggac atgccggact 1740ggttcacggc catcgtcatg gacaaggcgt acgacggctt
cgaacaggcc tactccttca 1800acgccacggg cgtcgggaac agcaccatct tcgtcagggg
cgcccagggc ctcaacttcc 1860tgctcgggga gaggaacatg agcggcgtag attacccggt
gcccgggaag cagcagtccg 1920tcttctcctt caccaagaag aagacccccg gcatcgacat
catcgccggg gacggcttcc 1980cgtccaaggt cttcttcaac ggcgacgagt gcgccatgcc
attgaggatt ccgagccagg 2040ggaccagtgt cgtcgtccct atgcagctgt gtttgcttgt
ttccgctttc atgttattgc 2100tgctgtaa
210816654PRTZea mays 16Met Ala Met Leu Pro Val Val
Val Arg Met Ala Ala Met Pro Phe Ile1 5 10
15Phe Leu Val Leu Leu Ala His Thr Ala Ser Ala Gln Pro
Asp Ala Gly 20 25 30Cys Asn
Gly Ile Leu Leu Thr Tyr Thr Leu Gln Arg Arg Asp Lys Ile 35
40 45Arg Pro His Val Ala Ala Pro Asn Ser Gln
Pro Tyr Ser Phe Ser Ala 50 55 60Ser
Ala Thr Val Val Asn Ala Gly Thr Arg Pro Leu Arg Ser Trp Ala65
70 75 80Leu Leu Leu Thr Phe Val
His Gly Glu Ile Leu Val Ser Val Asp Gly 85
90 95Ala Val Leu Thr Ser Gly Ala Ala Leu Pro Tyr Asn
Thr Thr Ala Gly 100 105 110Asp
Ala Ala Gly Arg Pro Thr Pro Thr Ser Phe Thr Gly Tyr Pro Gln 115
120 125Thr Asp Leu Leu Thr Pro Ile Ala Thr
Ala Gly Asp Pro Ala Lys Thr 130 135
140Gln Ala Thr Val Ser Leu Val Gly Thr Leu Phe Ala Gly Pro Glu Pro145
150 155 160Tyr Val Pro Leu
Pro Ser Phe Leu Ser Leu Ala Asp Pro Ser Tyr Thr 165
170 175Cys Pro Pro Ala Thr Asn Ala Thr Ser Ser
Pro Thr Asn Leu Thr Thr 180 185
190Cys Cys Val Phe Thr Ala Gly Gly Asp Pro Thr Gly Gly Leu Val Glu
195 200 205Ser Gly Phe Leu Pro Arg Arg
Thr Gly Asp Leu Val Ile Thr Tyr Asp 210 215
220Val Leu Gln Ser Tyr Asp Thr Thr Tyr Leu Ala Leu Val Thr Leu
Glu225 230 235 240Asn Asp
Ala Leu Leu Gly Arg Leu Asp Ala Trp Gln Leu Ser Trp Arg
245 250 255Trp Glu His Gly Glu Phe Ile
Ser Ser Met Arg Gly Ala Tyr Pro Arg 260 265
270Glu Val Asp Thr Ala Glu Cys Leu Tyr Gly Pro Gln Gly Gln
Tyr Tyr 275 280 285Lys Asp Leu Asp
Phe Ser Lys Ala Val Leu Asn Cys Asp Arg Arg Pro 290
295 300Val Val His Asp Leu Pro Pro Ser Arg Ala Asn Asp
Thr Glu Ile Gly305 310 315
320Arg Ile Asp His Cys Cys Arg Asn Gly Thr Ile Leu Pro Lys Ser Met
325 330 335Asp Val Ala Arg Ser
Lys Ser Ala Phe Gln Met Val Val Tyr Lys Met 340
345 350Pro Pro Asp Leu Asn Arg Thr Lys Leu Tyr Pro Pro
Thr Gly Phe Asn 355 360 365Val Thr
Gly Ala Ala Ser Ala Leu Asn Pro Glu Tyr Ala Cys Asp Pro 370
375 380Pro Ile Ser Val Ser Pro Ser Glu Tyr Pro Asp
Pro Ser Gly Leu Thr385 390 395
400Ser Ile Thr Val Ala Val Ala Thr Trp Gln Val Val Cys Asn Ile Thr
405 410 415Thr Ser Pro Lys
Lys Pro Pro Arg Cys Cys Val Ser Phe Ser Ser Phe 420
425 430Tyr Asn Glu Ser Val Val Pro Cys Arg Thr Cys
Ala Cys Gly Cys Pro 435 440 445Ser
Ser Ala Pro Thr Cys Ser Thr Thr Ala Pro Ala Met Leu Leu Pro 450
455 460Pro Gln Ala Leu Leu Met Pro Phe Asp Arg
Arg Ala Ser Glu Ala Leu465 470 475
480Glu Trp Ala Asp Gln Lys His Leu Gly Val Pro Lys Pro Met Pro
Cys 485 490 495Gly Asp Phe
Cys Gly Val Ser Val Asn Trp His Val Ala Thr Asp Phe 500
505 510Thr Gly Gly Trp Ser Ala Arg Leu Thr Leu
Phe Asn Trp Asp Gly Thr 515 520
525Asp Met Pro Asp Trp Phe Thr Ala Ile Val Met Asp Lys Ala Tyr Asp 530
535 540Gly Phe Glu Gln Ala Tyr Ser Phe
Asn Ala Thr Gly Val Gly Asn Ser545 550
555 560Thr Ile Phe Val Arg Gly Ala Gln Gly Leu Asn Phe
Leu Leu Gly Glu 565 570
575Arg Asn Met Ser Gly Val Asp Tyr Pro Val Pro Gly Lys Gln Gln Ser
580 585 590Val Phe Ser Phe Thr Lys
Lys Lys Thr Pro Gly Ile Asp Ile Ile Ala 595 600
605Gly Asp Gly Phe Pro Ser Lys Val Phe Phe Asn Gly Asp Glu
Cys Ala 610 615 620Met Pro Leu Arg Ile
Pro Ser Gln Gly Thr Ser Val Val Val Pro Met625 630
635 640Gln Leu Cys Leu Leu Val Ser Ala Phe Met
Leu Leu Leu Leu 645 650171335DNAZea mays
17atggggcgct tcgtcttcgt cctgctaatc ctgatgtgct gttcctcctc ccgattcaca
60ggtgcctacg accccataga tccgaacggg aacatcacga tcgtctggga ttttcagagt
120ctcgacgtcg cgggcatgac cccgtacacg gtcatggtga gcatccacaa ctaccagatg
180taccgacaca tcgagcgtcc ggggtggcgg ctgagctgga gctgggccgg caaggaggtc
240atctggagca cgacgggcgc ggagacgacg gagcagggcg actgctcccg cgtcggcagc
300gggggcagcc gcccgcattg ctgccagaag cggcccgtca tggtggacct gccgcctggc
360acgccgtaca acatgcaggt cgccaactgc tgccggggcg gcgtgctgtc gtctctcgtc
420cagagcgacc tgacgtccgc cgccgcgttc cagatggtgg tcggcgagtt cgccctcgcc
480agggacagcg gcggcaagga gcccgagaag ccgtggcagt tcgacatggg cgtgccgggg
540tacacctgca gcaacgcgac cacggtggcc ccgaccagga tcaaggtcga caagaaccgc
600tacgtccagg cgctccagga tcgagctgaa ctccgtgcag tgacatggca ggtgacctgc
660tcgtactcgc agtaccgggc gtcggcggcg ccgtcgtgct gcgtctccat gacgaccttc
720tacagcgaga cgatcgtaga ttgcccgcgg tgcagctgcg gctgccaagg gtccccaccg
780tcgccacaat gcgtcagcgt cgatcaacaa caaccatggt tgccggccgt cggcgacgac
840gagccgtcgt cggcgccgat cgtctggtgc tccgagcaca tgtgcccgat ccgggtgcac
900tggcacgtga agacgaacta ccgcaagtac tggcgggtga aggtgacggt gtccaactac
960aacctggcga ggaactacag cgactggaac ctggtgctgc agcaccccaa cctgcggagc
1020ctgacgcagc tgttcagctt caactacagg cctctcgtcg agtacggcgc ctacaacgac
1080acggggatgt tctgggggtt acgttactac aacgagatgc tgctgcagga cgggaacgtg
1140cagtcggaga tgatcctgga gaaggagagc gacttcacct attccggtgg ctgggcgttc
1200ccgcggaggg tctacttcaa cggccaagag tgcgtcatgc cgccggcgga ccagtacccc
1260gtactgccca acggagcctc ggctttgcgg gggcattttt gcttcctgct gctactcttc
1320ttcgttgtgg tgtag
133518444PRTZea mays 18Met Gly Arg Phe Val Phe Val Leu Leu Ile Leu Met
Cys Cys Ser Ser1 5 10
15Ser Arg Phe Thr Gly Ala Tyr Asp Pro Ile Asp Pro Asn Gly Asn Ile
20 25 30Thr Ile Val Trp Asp Phe Gln
Ser Leu Asp Val Ala Gly Met Thr Pro 35 40
45Tyr Thr Val Met Val Ser Ile His Asn Tyr Gln Met Tyr Arg His
Ile 50 55 60Glu Arg Pro Gly Trp Arg
Leu Ser Trp Ser Trp Ala Gly Lys Glu Val65 70
75 80Ile Trp Ser Thr Thr Gly Ala Glu Thr Thr Glu
Gln Gly Asp Cys Ser 85 90
95Arg Val Gly Ser Gly Gly Ser Arg Pro His Cys Cys Gln Lys Arg Pro
100 105 110Val Met Val Asp Leu Pro
Pro Gly Thr Pro Tyr Asn Met Gln Val Ala 115 120
125Asn Cys Cys Arg Gly Gly Val Leu Ser Ser Leu Val Gln Ser
Asp Leu 130 135 140Thr Ser Ala Ala Ala
Phe Gln Met Val Val Gly Glu Phe Ala Leu Ala145 150
155 160Arg Asp Ser Gly Gly Lys Glu Pro Glu Lys
Pro Trp Gln Phe Asp Met 165 170
175Gly Val Pro Gly Tyr Thr Cys Ser Asn Ala Thr Thr Val Ala Pro Thr
180 185 190Arg Ile Lys Val Asp
Lys Asn Arg Tyr Val Gln Ala Leu Gln Asp Arg 195
200 205Ala Glu Leu Arg Ala Val Thr Trp Gln Val Thr Cys
Ser Tyr Ser Gln 210 215 220Tyr Arg Ala
Ser Ala Ala Pro Ser Cys Cys Val Ser Met Thr Thr Phe225
230 235 240Tyr Ser Glu Thr Ile Val Asp
Cys Pro Arg Cys Ser Cys Gly Cys Gln 245
250 255Gly Ser Pro Pro Ser Pro Gln Cys Val Ser Val Asp
Gln Gln Gln Pro 260 265 270Trp
Leu Pro Ala Val Gly Asp Asp Glu Pro Ser Ser Ala Pro Ile Val 275
280 285Trp Cys Ser Glu His Met Cys Pro Ile
Arg Val His Trp His Val Lys 290 295
300Thr Asn Tyr Arg Lys Tyr Trp Arg Val Lys Val Thr Val Ser Asn Tyr305
310 315 320Asn Leu Ala Arg
Asn Tyr Ser Asp Trp Asn Leu Val Leu Gln His Pro 325
330 335Asn Leu Arg Ser Leu Thr Gln Leu Phe Ser
Phe Asn Tyr Arg Pro Leu 340 345
350Val Glu Tyr Gly Ala Tyr Asn Asp Thr Gly Met Phe Trp Gly Leu Arg
355 360 365Tyr Tyr Asn Glu Met Leu Leu
Gln Asp Gly Asn Val Gln Ser Glu Met 370 375
380Ile Leu Glu Lys Glu Ser Asp Phe Thr Tyr Ser Gly Gly Trp Ala
Phe385 390 395 400Pro Arg
Arg Val Tyr Phe Asn Gly Gln Glu Cys Val Met Pro Pro Ala
405 410 415Asp Gln Tyr Pro Val Leu Pro
Asn Gly Ala Ser Ala Leu Arg Gly His 420 425
430Phe Cys Phe Leu Leu Leu Leu Phe Phe Val Val Val
435 440193780DNAZea mays 19gtcgacccac gcgtccgcag
cagcagaagc actgcgcggc attgcagcga tcgagcggga 60ggaatttggg gcatggtggt
cgccaacgcc gctcggatct agaggcccgc acgggccgat 120tggtctccgc ccgcctcgtc
ggtgttggtg tcgttggcgt gtggagccgt ctcggtggga 180gcagcgggga gggagcggag
atggcggcca acaaggggat ggtggcgggc tcgcacaacc 240gcaacgagtt cgtcatgatc
cgccacgacg gcgatgtgcc gggctcggct aagcccacaa 300agagtgcgaa tggacaggtc
tgccagattt gcggtgactc tgtgggtgtt tcagccactg 360gtgatgtctt tgttgcctgc
aatgagtgtg ccttccctgt ctgccgccca tgctatgagt 420atgagcgcaa ggaggggaac
caatgctgcc cccagtgcaa gactagatac aagagacaga 480aaggtagccc tcgagttcat
ggtgatgagg atgaggaaga tgttgatgac ctagacaatg 540aattcaacta caagcaaggc
agtgggaaag gcccagagtg gcaactgcaa ggagatgatg 600ctgatctgtc ttcatctgct
cgccatgagc cacatcatcg gattccacgc ctgacaagcg 660gtcaacagat atctggagag
attcctgatg cttcccctga ccgtcattct atccgcagtc 720caacatcgag ctatgttgat
ccaagcgtcc cagttcctgt gaggattgtg gacccctcga 780aggacttgaa ttcctatggg
cttaatagtg ttgactggaa ggaaagagtt gagagctgga 840gggttaaaca ggacaaaaat
atgatgcaag tgactaataa atatccagag gctagaggag 900gagacatgga ggggactggc
tcaaatggag aagatatgca aatggttgat gatgcacggc 960tacctttgag ccgtatcgtg
ccaatttcct caaaccagct caacctttac cgggtagtga 1020tcattctccg tcttatcatc
ctgtgcttct tcttccagta tcgtgtcagt catccagtgc 1080gtgatgctta tggattatgg
ctagtatctg ttatctgcga ggtctggttt gccttgtctt 1140ggcttctaga tcagttccca
aaatggtatc caatcaaccg tgagacatat cttgacaggc 1200ttgcattgag gtatgataga
gagggagagc catcacagct ggctcccatt gatgtcttcg 1260tcagtacagt ggatccattg
aaggaacctc cactgatcac agccaacact gttttgtcca 1320ttctttctgt ggattaccct
gttgacaaag tgtcatgcta tgtttctgat gatggttcag 1380ctatgctgac ttttgagtct
ctctcagaaa ccgcagaatt tgctagaaag tgggttccct 1440tttgtaagaa gcacaatatt
gaaccaagag ctccagaatt ttactttgct caaaaaatag 1500attacctgaa ggacaaaatt
caaccttcat ttgttaagga aagacgcgca atgaagaggg 1560agtatgaaga attcaaagta
agaatcaatg cccttgttgc caaagcacag aaagtgcctg 1620aagaggggtg gaccatggct
gatggaactg catggcctgg gaataatcct agggaccatc 1680ctggcatgat tcaggttttc
ttggggcaca gtggtgggct cgacactgat ggaaatgagt 1740taccacgtct tgtctatgtc
tctcgtgaaa agagaccagg ctttcagcat cacaagaagg 1800ctggtgcaat gaatgcgctg
attcgtgtat ctgctgtgct gacaaatggt gcctatcttc 1860tcaatgtgga ttgcgaccat
tacttcaata gcagcaaagc tcttagagaa gcaatgtgct 1920tcatgatgga tccggctcta
ggaaggaaaa cttgttatgt acaatttcca cagagatttg 1980atggcattga cttgcacgat
cgatatgcta atcggaacat agttttcttt gatatcaaca 2040tgaaaggtct ggatggcatt
cagggtccag tttacgtggg aacaggatgc tgtttcaata 2100gacaggcttt gtatggatac
gatcctgttt tgactgaagc tgatctggag ccaaacattg 2160ttattaagag ctgctgtggt
agaaggaaga aaaagaacaa gagttatatg gatagtcaaa 2220gccgtattat gaagagaaca
gaatcttcag ctcccatctt caatatggaa gacatcgaag 2280agggtattga aggttacgag
gatgaaaggt cagtgcttat gtcccagagg aaattggaga 2340aacgctttgg tcagtctcct
attttcattg catccacctt tatgacacaa ggtggcatac 2400caccttcaac aaacccagct
tctctactaa aggaagctat ccatgtcatc agttgtggat 2460atgaggacaa aactgaatgg
ggaaaagaga ttggctggat ctatggttca gtaacggagg 2520atattctgac tgggtttaaa
atgcatgcaa ggggctggca atcaatctac tgcatgccac 2580cacgaccttg tttcaagggt
tctgcaccaa tcaatctttc cgatcgtctt aatcaggtgc 2640tccgttgggc tcttgggtca
gtggaaattc tgcttagtag acattgtcct atctggtatg 2700gttacaatgg acgattgaag
cttttggaga ggctggctta catcaacact attgtatatc 2760caatcacatc cattccgctt
attgcctatt gtgtgcttcc cgctatctgc ctccttacca 2820ataaatttat cattcctgag
attagcaatt atgctgggat gttcttcatt cttcttttcg 2880cctccatttt tgccactggt
atattggagc ttagatggag tggtgttggc attgaagatt 2940ggtggagaaa tgagcagttt
tgggttattg gtggcacctc tgcccatctc ttcgcagtgt 3000tccagggtct gctgaaagtg
ttggctggga ttgataccaa cttcacagtt acctcaaagg 3060catctgatga ggatggcgac
tttgctgagc tatatgtgtt caagtggacc agtttgctca 3120ttcctccgac cactgttctt
gtcattaacc tggtcggaat ggtggcagga atttcttatg 3180ccattaacag tggctaccaa
tcctggggtc cgctctttgg aaagctgttc ttctcgatct 3240gggtgatcct ccatctctac
cccttcctca agggtctcat gggaaggcag aaccgcacac 3300caacaatcgt cattgtctgg
tccatccttc ttgcatctat cttctccttg ctgtgggtga 3360agatcgatcc tttcatctcc
ccgacacaga aagctgctgc cttggggcaa tgtggcgtca 3420actgctgatc gagacagtga
ctcttatttg aagaggctca atcaagatct gccccctcgt 3480gtaaatacct gaggaggcta
gatgggaatt ccttttgttg taggtgagga tggatttgca 3540tctaagttat gcctctgttc
attagcttct tccgtgccgg tgctgctgcg gactaagaat 3600cacggagcct ttctaccttc
catgtagcgc cagccagcag cgtaagatgt gaattttgaa 3660gttttgttat gcgtgcagtt
tattgtttta gagtaaatta tcatttgttt gtgggaactg 3720ttcacacgag cttataatgg
caatgctgtt atttaaaaaa aaaaaaaaaa gggcggccgc 3780201075PRTZea mays 20Met
Ala Ala Asn Lys Gly Met Val Ala Gly Ser His Asn Arg Asn Glu1
5 10 15Phe Val Met Ile Arg His Asp
Gly Asp Val Pro Gly Ser Ala Lys Pro 20 25
30Thr Lys Ser Ala Asn Gly Gln Val Cys Gln Ile Cys Gly Asp
Ser Val 35 40 45Gly Val Ser Ala
Thr Gly Asp Val Phe Val Ala Cys Asn Glu Cys Ala 50 55
60Phe Pro Val Cys Arg Pro Cys Tyr Glu Tyr Glu Arg Lys
Glu Gly Asn65 70 75
80Gln Cys Cys Pro Gln Cys Lys Thr Arg Tyr Lys Arg Gln Lys Gly Ser
85 90 95Pro Arg Val His Gly Asp
Glu Asp Glu Glu Asp Val Asp Asp Leu Asp 100
105 110Asn Glu Phe Asn Tyr Lys Gln Gly Ser Gly Lys Gly
Pro Glu Trp Gln 115 120 125Leu Gln
Gly Asp Asp Ala Asp Leu Ser Ser Ser Ala Arg His Glu Pro 130
135 140His His Arg Ile Pro Arg Leu Thr Ser Gly Gln
Gln Ile Ser Gly Glu145 150 155
160Ile Pro Asp Ala Ser Pro Asp Arg His Ser Ile Arg Ser Pro Thr Ser
165 170 175Ser Tyr Val Asp
Pro Ser Val Pro Val Pro Val Arg Ile Val Asp Pro 180
185 190Ser Lys Asp Leu Asn Ser Tyr Gly Leu Asn Ser
Val Asp Trp Lys Glu 195 200 205Arg
Val Glu Ser Trp Arg Val Lys Gln Asp Lys Asn Met Met Gln Val 210
215 220Thr Asn Lys Tyr Pro Glu Ala Arg Gly Gly
Asp Met Glu Gly Thr Gly225 230 235
240Ser Asn Gly Glu Asp Met Gln Met Val Asp Asp Ala Arg Leu Pro
Leu 245 250 255Ser Arg Ile
Val Pro Ile Ser Ser Asn Gln Leu Asn Leu Tyr Arg Val 260
265 270Val Ile Ile Leu Arg Leu Ile Ile Leu Cys
Phe Phe Phe Gln Tyr Arg 275 280
285Val Ser His Pro Val Arg Asp Ala Tyr Gly Leu Trp Leu Val Ser Val 290
295 300Ile Cys Glu Val Trp Phe Ala Leu
Ser Trp Leu Leu Asp Gln Phe Pro305 310
315 320Lys Trp Tyr Pro Ile Asn Arg Glu Thr Tyr Leu Asp
Arg Leu Ala Leu 325 330
335Arg Tyr Asp Arg Glu Gly Glu Pro Ser Gln Leu Ala Pro Ile Asp Val
340 345 350Phe Val Ser Thr Val Asp
Pro Leu Lys Glu Pro Pro Leu Ile Thr Ala 355 360
365Asn Thr Val Leu Ser Ile Leu Ser Val Asp Tyr Pro Val Asp
Lys Val 370 375 380Ser Cys Tyr Val Ser
Asp Asp Gly Ser Ala Met Leu Thr Phe Glu Ser385 390
395 400Leu Ser Glu Thr Ala Glu Phe Ala Arg Lys
Trp Val Pro Phe Cys Lys 405 410
415Lys His Asn Ile Glu Pro Arg Ala Pro Glu Phe Tyr Phe Ala Gln Lys
420 425 430Ile Asp Tyr Leu Lys
Asp Lys Ile Gln Pro Ser Phe Val Lys Glu Arg 435
440 445Arg Ala Met Lys Arg Glu Tyr Glu Glu Phe Lys Val
Arg Ile Asn Ala 450 455 460Leu Val Ala
Lys Ala Gln Lys Val Pro Glu Glu Gly Trp Thr Met Ala465
470 475 480Asp Gly Thr Ala Trp Pro Gly
Asn Asn Pro Arg Asp His Pro Gly Met 485
490 495Ile Gln Val Phe Leu Gly His Ser Gly Gly Leu Asp
Thr Asp Gly Asn 500 505 510Glu
Leu Pro Arg Leu Val Tyr Val Ser Arg Glu Lys Arg Pro Gly Phe 515
520 525Gln His His Lys Lys Ala Gly Ala Met
Asn Ala Leu Ile Arg Val Ser 530 535
540Ala Val Leu Thr Asn Gly Ala Tyr Leu Leu Asn Val Asp Cys Asp His545
550 555 560Tyr Phe Asn Ser
Ser Lys Ala Leu Arg Glu Ala Met Cys Phe Met Met 565
570 575Asp Pro Ala Leu Gly Arg Lys Thr Cys Tyr
Val Gln Phe Pro Gln Arg 580 585
590Phe Asp Gly Ile Asp Leu His Asp Arg Tyr Ala Asn Arg Asn Ile Val
595 600 605Phe Phe Asp Ile Asn Met Lys
Gly Leu Asp Gly Ile Gln Gly Pro Val 610 615
620Tyr Val Gly Thr Gly Cys Cys Phe Asn Arg Gln Ala Leu Tyr Gly
Tyr625 630 635 640Asp Pro
Val Leu Thr Glu Ala Asp Leu Glu Pro Asn Ile Val Ile Lys
645 650 655Ser Cys Cys Gly Arg Arg Lys
Lys Lys Asn Lys Ser Tyr Met Asp Ser 660 665
670Gln Ser Arg Ile Met Lys Arg Thr Glu Ser Ser Ala Pro Ile
Phe Asn 675 680 685Met Glu Asp Ile
Glu Glu Gly Ile Glu Gly Tyr Glu Asp Glu Arg Ser 690
695 700Val Leu Met Ser Gln Arg Lys Leu Glu Lys Arg Phe
Gly Gln Ser Pro705 710 715
720Ile Phe Ile Ala Ser Thr Phe Met Thr Gln Gly Gly Ile Pro Pro Ser
725 730 735Thr Asn Pro Ala Ser
Leu Leu Lys Glu Ala Ile His Val Ile Ser Cys 740
745 750Gly Tyr Glu Asp Lys Thr Glu Trp Gly Lys Glu Ile
Gly Trp Ile Tyr 755 760 765Gly Ser
Val Thr Glu Asp Ile Leu Thr Gly Phe Lys Met His Ala Arg 770
775 780Gly Trp Gln Ser Ile Tyr Cys Met Pro Pro Arg
Pro Cys Phe Lys Gly785 790 795
800Ser Ala Pro Ile Asn Leu Ser Asp Arg Leu Asn Gln Val Leu Arg Trp
805 810 815Ala Leu Gly Ser
Val Glu Ile Leu Leu Ser Arg His Cys Pro Ile Trp 820
825 830Tyr Gly Tyr Asn Gly Arg Leu Lys Leu Leu Glu
Arg Leu Ala Tyr Ile 835 840 845Asn
Thr Ile Val Tyr Pro Ile Thr Ser Ile Pro Leu Ile Ala Tyr Cys 850
855 860Val Leu Pro Ala Ile Cys Leu Leu Thr Asn
Lys Phe Ile Ile Pro Glu865 870 875
880Ile Ser Asn Tyr Ala Gly Met Phe Phe Ile Leu Leu Phe Ala Ser
Ile 885 890 895Phe Ala Thr
Gly Ile Leu Glu Leu Arg Trp Ser Gly Val Gly Ile Glu 900
905 910Asp Trp Trp Arg Asn Glu Gln Phe Trp Val
Ile Gly Gly Thr Ser Ala 915 920
925His Leu Phe Ala Val Phe Gln Gly Leu Leu Lys Val Leu Ala Gly Ile 930
935 940Asp Thr Asn Phe Thr Val Thr Ser
Lys Ala Ser Asp Glu Asp Gly Asp945 950
955 960Phe Ala Glu Leu Tyr Val Phe Lys Trp Thr Ser Leu
Leu Ile Pro Pro 965 970
975Thr Thr Val Leu Val Ile Asn Leu Val Gly Met Val Ala Gly Ile Ser
980 985 990Tyr Ala Ile Asn Ser Gly
Tyr Gln Ser Trp Gly Pro Leu Phe Gly Lys 995 1000
1005Leu Phe Phe Ser Ile Trp Val Ile Leu His Leu Tyr
Pro Phe Leu 1010 1015 1020Lys Gly Leu
Met Gly Arg Gln Asn Arg Thr Pro Thr Ile Val Ile 1025
1030 1035Val Trp Ser Ile Leu Leu Ala Ser Ile Phe Ser
Leu Leu Trp Val 1040 1045 1050Lys Ile
Asp Pro Phe Ile Ser Pro Thr Gln Lys Ala Ala Ala Leu 1055
1060 1065Gly Gln Cys Gly Val Asn Cys 1070
1075213725DNAZea mays 21gcagcagcag caccaccact gcgcggcatt
gcagcgagca agcgggaggg atctggggca 60tggtggcggt cgctgccgct gccgctcgga
tctagagggc cgcacgggct gattgccctc 120cgccggcctc gtcggtgtcg gtggagtgtg
aatcggtgtg tgtaggagga gcgcggagat 180ggcggccaac aaggggatgg tggcaggctc
tcacaaccgc aacgagttcg tcatgatccg 240ccacgacggc gacgcgcctg tcccggctaa
gcccacgaag agtgcgaatg ggcaggtctg 300ccagatttgt ggcgacactg ttggcgtttc
agccactggt gatgtctttg ttgcctgcaa 360tgagtgtgcc ttccctgtct gccgcccttg
ctatgagtac gagcgcaagg aagggaacca 420atgctgccct cagtgcaaga ctagatacaa
gagacagaaa ggtagccctc gagttcatgg 480tgatgatgag gaggaagatg ttgatgacct
ggacaatgaa ttcaactata agcaaggcaa 540tgggaagggc ccagagtggc agcttcaagg
agatgacgct gatctgtctt catctgctcg 600ccatgaccca caccatcgga ttccacgcct
tacaagtgga caacagatat ctggagagat 660ccctgatgca tcccctgacc gtcattctat
ccgcagtcca acatcgagct atgttgatcc 720aagcgttcca gttcctgtga ggattgtgga
cccctcgaag gacttgaatt cctatgggct 780taatagtgtt gactggaagg aaagagttga
gagctggagg gttaaacagg acaaaaatat 840gttgcaagtg actaataaat atccagaggc
tagaggagac atggagggga ctggctcaaa 900tggagaagat atgcaaatgg ttgatgatgc
acgcctacct ttgagccgca ttgtgccaat 960ttcctcaaac cagctcaacc tttaccggat
agtaatcatt ctccgtctta tcatcctgtg 1020cttcttcttc caatatcgta tcagtcatcc
agtgcgtaat gcttatggat tgtggctagt 1080atctgttatc tgtgaggtct ggtttgcctt
gtcctggctt ctagatcagt tcccaaaatg 1140gtatccaatc aaccgtgaga catatctcga
caggcttgca ttgaggtatg atagagaggg 1200agagccatca cagctggctc ccattgatgt
ctttgtcagt acagtggatc cattgaagga 1260acctccactg atcacagcca acactgtttt
gtccattctt gctgtggatt accctgttga 1320caaagtgtca tgctatgttt ctgatgatgg
ctcagctatg ctgacttttg agtctctctc 1380tgaaactgcc gaatttgcta gaaagtgggt
tcccttttgt aagaagcaca atattgaacc 1440aagagctcca gaattttact ttgctcaaaa
aatagattac ctgaaggaca aaattcaacc 1500ttcatttgtt aaggaaagac gagcaatgaa
gagagagtat gaagaattca aaataagaat 1560caatgccctt gttgccaaag cacagaaagt
gcctgaagag gggtggacca tggctgatgg 1620aactgcttgg cctgggaata accctaggga
ccatcctggc atgattcagg tgttcttggg 1680gcacagtggt gggcttgaca ctgatggaaa
tgaattacca cgtcttgtct atgtctctcg 1740tgaaaagaga ccaggctttc agcatcacaa
gaaggctggt gcaatgaatg cactgattcg 1800tgtatctgct gtgctgacaa atggtgccta
tcttctcaat gtggattgtg accattactt 1860caatagcagc aaagctctta gagaagcaat
gtgcttcatg atggatccag ctctaggaag 1920gaaaacttgt tatgtacaat ttccacaaag
atttgatggc attgacttgc acgatcgata 1980tgctaatagg aacatagtct tctttgatat
caacatgaaa ggtctagatg gcattcaggg 2040tccagtctat gtgggaacag gatgctgttt
caataggcag gctttgtatg gatatgatcc 2100tgttttgact gaagctgatc tggaacctaa
cattgttgtt aagagctgct gtggtagaag 2160gaagagaaag aacaagagtt atatggatag
tcaaagccgt attatgaaga gaacagaatc 2220ttcagctccc atctttaaca tggaagacat
cgaggagggt attgaaggtt atgaggatga 2280aaggtcagtg cttatgtccc agaggaaatt
ggagaaacgc tttggtcagt ctccaatctt 2340cattgcatcc acctttatga ctcaaggtgg
cataccacct tcaacaaacc cagcttctct 2400actgaaggaa gctatccatg ttatcagctg
tgggtacgag gacaaaactg aatggggaaa 2460agagattggc tggatctatg gttcagttac
agaggatatt ctgactgggt ttaaaatgca 2520tgcaagaggc tggcaatcaa tctactgcat
gccaccacga ccttgtttca agggttctgc 2580accaatcaat ctttctgatc gtcttaatca
ggtgctccgt tgggctcttg ggtcagtgga 2640aattctgctt agcagacatt gtcctatatg
gtatggctac aatgggcgat tgaagctttt 2700ggagaggctg gcttacatta acaccattgt
ttatccaatc acatctgttc cgcttatcgc 2760ctattgtgtg cttcctgcta tctgtcttct
taccaataaa tttatcattc ctgagattag 2820taattatgct ggaatgttct tcattcttct
ttttgcctcc attttcgcaa ctggtatatt 2880ggagctcaga tggagtggtg ttggcattga
agattggtgg agaaatgagc agttttgggt 2940tattggtggc acctctgccc atctcttcgc
ggtgttccag ggtctgctga aagtgttggc 3000tgggattgat accaacttca cagttacctc
aaaggcatct gatgaggatg gcgactttgc 3060tgagctatat gtgttcaagt ggaccagttt
gctcatccct ccgaccactg ttcttgtcat 3120taacctggtc ggaatggtgg caggaatttc
gtatgccatt aacagcggct accaatcctg 3180gggtccgctc tttggaaagc tgttcttctc
gatctgggtg atcctccatc tctacccctt 3240cctcaagggt ctcatgggca ggcagaaccg
cacgccaaca atcgtcatcg tttggtccat 3300cctccttgcg tctatcttct ccttgctgtg
ggtgaagatc gatcctttca tctccccgac 3360acagaaagct gccgccttgg ggcaatgtgg
tgtgaactgc tgatccagat tgtgactctt 3420atctgaagag gctcagccaa agatctgccc
cctcgtgtaa atacctgagg gggctagatg 3480ggaatttttt gttgtagatg aggatggatc
tgcatccaag ttatgcctct gtttattagc 3540ttcttcggtg ccggtgctgc tgcagacaat
catggagcct ttctaccttg cttgtagtgc 3600tggccagcag cgtaaattgt gaattctgca
tttttttata cgtggtgttt attgttttag 3660agtaaattat catttgtttg aggtaactat
tcacacgaac tatatggcaa tgctgttatt 3720taaaa
3725221074PRTZea mays 22Met Ala Ala Asn
Lys Gly Met Val Ala Gly Ser His Asn Arg Asn Glu1 5
10 15Phe Val Met Ile Arg His Asp Gly Asp Ala
Pro Val Pro Ala Lys Pro 20 25
30Thr Lys Ser Ala Asn Gly Gln Val Cys Gln Ile Cys Gly Asp Thr Val
35 40 45Gly Val Ser Ala Thr Gly Asp Val
Phe Val Ala Cys Asn Glu Cys Ala 50 55
60Phe Pro Val Cys Arg Pro Cys Tyr Glu Tyr Glu Arg Lys Glu Gly Asn65
70 75 80Gln Cys Cys Pro Gln
Cys Lys Thr Arg Tyr Lys Arg Gln Lys Gly Ser 85
90 95Pro Arg Val His Gly Asp Asp Glu Glu Glu Asp
Val Asp Asp Leu Asp 100 105
110Asn Glu Phe Asn Tyr Lys Gln Gly Asn Gly Lys Gly Pro Glu Trp Gln
115 120 125Leu Gln Gly Asp Asp Ala Asp
Leu Ser Ser Ser Ala Arg His Asp Pro 130 135
140His His Arg Ile Pro Arg Leu Thr Ser Gly Gln Gln Ile Ser Gly
Glu145 150 155 160Ile Pro
Asp Ala Ser Pro Asp Arg His Ser Ile Arg Ser Pro Thr Ser
165 170 175Ser Tyr Val Asp Pro Ser Val
Pro Val Pro Val Arg Ile Val Asp Pro 180 185
190Ser Lys Asp Leu Asn Ser Tyr Gly Leu Asn Ser Val Asp Trp
Lys Glu 195 200 205Arg Val Glu Ser
Trp Arg Val Lys Gln Asp Lys Asn Met Leu Gln Val 210
215 220Thr Asn Lys Tyr Pro Glu Ala Arg Gly Asp Met Glu
Gly Thr Gly Ser225 230 235
240Asn Gly Glu Asp Met Gln Met Val Asp Asp Ala Arg Leu Pro Leu Ser
245 250 255Arg Ile Val Pro Ile
Ser Ser Asn Gln Leu Asn Leu Tyr Arg Ile Val 260
265 270Ile Ile Leu Arg Leu Ile Ile Leu Cys Phe Phe Phe
Gln Tyr Arg Ile 275 280 285Ser His
Pro Val Arg Asn Ala Tyr Gly Leu Trp Leu Val Ser Val Ile 290
295 300Cys Glu Val Trp Phe Ala Leu Ser Trp Leu Leu
Asp Gln Phe Pro Lys305 310 315
320Trp Tyr Pro Ile Asn Arg Glu Thr Tyr Leu Asp Arg Leu Ala Leu Arg
325 330 335Tyr Asp Arg Glu
Gly Glu Pro Ser Gln Leu Ala Pro Ile Asp Val Phe 340
345 350Val Ser Thr Val Asp Pro Leu Lys Glu Pro Pro
Leu Ile Thr Ala Asn 355 360 365Thr
Val Leu Ser Ile Leu Ala Val Asp Tyr Pro Val Asp Lys Val Ser 370
375 380Cys Tyr Val Ser Asp Asp Gly Ser Ala Met
Leu Thr Phe Glu Ser Leu385 390 395
400Ser Glu Thr Ala Glu Phe Ala Arg Lys Trp Val Pro Phe Cys Lys
Lys 405 410 415His Asn Ile
Glu Pro Arg Ala Pro Glu Phe Tyr Phe Ala Gln Lys Ile 420
425 430Asp Tyr Leu Lys Asp Lys Ile Gln Pro Ser
Phe Val Lys Glu Arg Arg 435 440
445Ala Met Lys Arg Glu Tyr Glu Glu Phe Lys Ile Arg Ile Asn Ala Leu 450
455 460Val Ala Lys Ala Gln Lys Val Pro
Glu Glu Gly Trp Thr Met Ala Asp465 470
475 480Gly Thr Ala Trp Pro Gly Asn Asn Pro Arg Asp His
Pro Gly Met Ile 485 490
495Gln Val Phe Leu Gly His Ser Gly Gly Leu Asp Thr Asp Gly Asn Glu
500 505 510Leu Pro Arg Leu Val Tyr
Val Ser Arg Glu Lys Arg Pro Gly Phe Gln 515 520
525His His Lys Lys Ala Gly Ala Met Asn Ala Leu Ile Arg Val
Ser Ala 530 535 540Val Leu Thr Asn Gly
Ala Tyr Leu Leu Asn Val Asp Cys Asp His Tyr545 550
555 560Phe Asn Ser Ser Lys Ala Leu Arg Glu Ala
Met Cys Phe Met Met Asp 565 570
575Pro Ala Leu Gly Arg Lys Thr Cys Tyr Val Gln Phe Pro Gln Arg Phe
580 585 590Asp Gly Ile Asp Leu
His Asp Arg Tyr Ala Asn Arg Asn Ile Val Phe 595
600 605Phe Asp Ile Asn Met Lys Gly Leu Asp Gly Ile Gln
Gly Pro Val Tyr 610 615 620Val Gly Thr
Gly Cys Cys Phe Asn Arg Gln Ala Leu Tyr Gly Tyr Asp625
630 635 640Pro Val Leu Thr Glu Ala Asp
Leu Glu Pro Asn Ile Val Val Lys Ser 645
650 655Cys Cys Gly Arg Arg Lys Arg Lys Asn Lys Ser Tyr
Met Asp Ser Gln 660 665 670Ser
Arg Ile Met Lys Arg Thr Glu Ser Ser Ala Pro Ile Phe Asn Met 675
680 685Glu Asp Ile Glu Glu Gly Ile Glu Gly
Tyr Glu Asp Glu Arg Ser Val 690 695
700Leu Met Ser Gln Arg Lys Leu Glu Lys Arg Phe Gly Gln Ser Pro Ile705
710 715 720Phe Ile Ala Ser
Thr Phe Met Thr Gln Gly Gly Ile Pro Pro Ser Thr 725
730 735Asn Pro Ala Ser Leu Leu Lys Glu Ala Ile
His Val Ile Ser Cys Gly 740 745
750Tyr Glu Asp Lys Thr Glu Trp Gly Lys Glu Ile Gly Trp Ile Tyr Gly
755 760 765Ser Val Thr Glu Asp Ile Leu
Thr Gly Phe Lys Met His Ala Arg Gly 770 775
780Trp Gln Ser Ile Tyr Cys Met Pro Pro Arg Pro Cys Phe Lys Gly
Ser785 790 795 800Ala Pro
Ile Asn Leu Ser Asp Arg Leu Asn Gln Val Leu Arg Trp Ala
805 810 815Leu Gly Ser Val Glu Ile Leu
Leu Ser Arg His Cys Pro Ile Trp Tyr 820 825
830Gly Tyr Asn Gly Arg Leu Lys Leu Leu Glu Arg Leu Ala Tyr
Ile Asn 835 840 845Thr Ile Val Tyr
Pro Ile Thr Ser Val Pro Leu Ile Ala Tyr Cys Val 850
855 860Leu Pro Ala Ile Cys Leu Leu Thr Asn Lys Phe Ile
Ile Pro Glu Ile865 870 875
880Ser Asn Tyr Ala Gly Met Phe Phe Ile Leu Leu Phe Ala Ser Ile Phe
885 890 895Ala Thr Gly Ile Leu
Glu Leu Arg Trp Ser Gly Val Gly Ile Glu Asp 900
905 910Trp Trp Arg Asn Glu Gln Phe Trp Val Ile Gly Gly
Thr Ser Ala His 915 920 925Leu Phe
Ala Val Phe Gln Gly Leu Leu Lys Val Leu Ala Gly Ile Asp 930
935 940Thr Asn Phe Thr Val Thr Ser Lys Ala Ser Asp
Glu Asp Gly Asp Phe945 950 955
960Ala Glu Leu Tyr Val Phe Lys Trp Thr Ser Leu Leu Ile Pro Pro Thr
965 970 975Thr Val Leu Val
Ile Asn Leu Val Gly Met Val Ala Gly Ile Ser Tyr 980
985 990Ala Ile Asn Ser Gly Tyr Gln Ser Trp Gly Pro
Leu Phe Gly Lys Leu 995 1000
1005Phe Phe Ser Ile Trp Val Ile Leu His Leu Tyr Pro Phe Leu Lys
1010 1015 1020Gly Leu Met Gly Arg Gln
Asn Arg Thr Pro Thr Ile Val Ile Val 1025 1030
1035Trp Ser Ile Leu Leu Ala Ser Ile Phe Ser Leu Leu Trp Val
Lys 1040 1045 1050Ile Asp Pro Phe Ile
Ser Pro Thr Gln Lys Ala Ala Ala Leu Gly 1055 1060
1065Gln Cys Gly Val Asn Cys 1070232830DNAZea
maysmisc_feature(2809)..(2809)n is a, c, g, or t 23tacctctaag tcgcatagtt
ccgatatctc caaacgagct taacctttat cggatcgtga 60ttgttctccg gcttatcatc
ctatgtttct tctttcaata tcgtataact catccagtgg 120aagatgctta tgggttgtgg
cttgtatctg ttatttgtga agtttggttt gccttgtctt 180ggcttctaga tcagttccca
aagtggtatc ctatcaaccg tgaaacttac ctcgatagac 240ttgcattgag atatgatagg
gagggtgagc catcccagtt ggctccaatc gatgtctttg 300ttagtacagt ggatccactt
aaggaacctc ctctaattac tggcaacact gtcctgtcca 360ttcttgctgt ggattaccct
gttgacaaag tatcatgtta tgtttctgat gacggttcag 420ctatgttgac ttttgaagcg
ctatctgaaa ccgcagagtt tgcaaggaaa tgggttccct 480tttgcaagaa acacaatatt
gaacctaggg ctccagagtt ttactttgct cgaaagatag 540attacctaaa ggacaaaata
caaccttctt ttgtgaaaga aaggcgggct atgaagaggg 600agtgtgaaga gttcaaagta
cggatcgatg cccttgttgc aaaagcgcaa aaaatacctg 660aggagggctg gaccatggct
gatggcactc cttggcctgg gaataaccct agagatcatc 720caggaatgat ccaagtattc
ttgggccaca gtggtgggct tgacacggat gggaatgagt 780tgccacggct tgtttatgtt
tctcgtgaaa agaggccagg cttccagcac cacaagaagg 840ctggtgccat gaatgctttg
attcgcgtat cagctgtcct gacgaatggt gcttatcttc 900ttaatgtgga ttgtgatcac
tacttcaata gcagcaaagc tcttagagag gctatgtgtt 960tcatgatgga tccagcacta
ggaaggaaaa cttgctatgt tcagtttcca caaagatttg 1020atggtataga cttgcatgat
cgatatgcaa accggaacat tgtcttcttt gatattaata 1080tgaagggtct agatggcatt
caaggacctg tttatgtggg aacaggatgc tgtttcaata 1140ggcaggcctt gtatggctat
gatcctgtat tgacagaagc tgatttggag cctaacatta 1200tcattaaaag ttgctgtggc
ggaagaaaaa agaaggacaa gagctatatt gattccaaaa 1260accgtgatat gaagagaaca
gaatcttcgg ctcccatctt caacatggaa gatatagaag 1320agggatttga aggttacgag
gatgaaaggt cactgcttat gtctcagaag agcttggaga 1380aacgctttgg ccagtctcca
atttttattg catccacctt tatgactcaa ggtggcatac 1440ccccttcaac aaacccaggt
tccctgctaa aggaagctat acatgtcatt agttgtggat 1500atgaggataa aacagaatgg
gggaaagaga tcggatggat atatggctct gttactgaag 1560atattttaac tggtttcaag
atgcatgcaa gaggttggat atccatctac tgcatgccac 1620ttcggccttg cttcaagggt
tctgctccaa ttaatctttc tgatcgtctc aaccaagtgt 1680tacgctgggc tcttggttca
gttgaaattc tacttagcag acactgtcct atctggtatg 1740gttacaatgg aaggctaaag
cttctggaga gactggcata catcaacacc attgtttatc 1800caattacatc tatcccacta
gtagcatact gcgtccttcc tgctatctgt ttactcacca 1860acaaatttat tattcctgcg
attagcaatt atgctggggc gttcttcatc ctgctttttg 1920cttccatctt cgccactggt
attttggagc ttcgatggag tggtgttggc attgaggatt 1980ggtggagaaa tgagcagttt
tgggtcattg gtggcacctc tgcacatctc tttgctgtgt 2040tccaaggtct cttaaaagtg
ctagcaggga tcgacacaaa cttcacggtc acatcaaagg 2100caaccgatga tgatggtgat
tttgctgagc tgtatgtgtt caagtggaca actcttctga 2160tcccccccac cactgtgctt
gtgattaacc tggttggtat agtggctgga gtgtcgtatg 2220ctatcaacag tggctaccaa
tcatggggtc cactattcgg gaagctgttc tttgcaatct 2280gggtgatcct ccacctctac
cctttcctga agggtctcat ggggaagcag aaccgcacac 2340cgaccatcgt catcgtttgg
tccgtccttc ttgcttccat attctcgctg ctgtgggtga 2400agatcgaccc cttcatatcc
cctacccaga aggctctttc ccgtgggcag tgtggtgtaa 2460actgctgaaa tgatccgaac
tgcctgctga ataacattgc tccggcacaa tcatgatcta 2520ccccttcgtg taaataccag
aggttaggca agacttttct tggtaggtgg cgaagatgtg 2580tcgtttaagt tcactctact
gcatttgggg tgggcagcat gaaactttgt caacttatgt 2640cgtgctactt atttgtagct
aagtagcagt aagtagtgcc tgtttcatgt tgactgtcgt 2700gactacctgt tcaccgtggg
ctctggactg tcgtgatgta acctgtatgt tggaacttca 2760agtactgatt gagctgtttg
gtcaatgaca ttgagggatt ctctctctng aaattaanac 2820aaantnggnt
283024821PRTZea mays 24Pro
Leu Ser Arg Ile Val Pro Ile Ser Pro Asn Glu Leu Asn Leu Tyr1
5 10 15Arg Ile Val Ile Val Leu Arg
Leu Ile Ile Leu Cys Phe Phe Phe Gln 20 25
30Tyr Arg Ile Thr His Pro Val Glu Asp Ala Tyr Gly Leu Trp
Leu Val 35 40 45Ser Val Ile Cys
Glu Val Trp Phe Ala Leu Ser Trp Leu Leu Asp Gln 50 55
60Phe Pro Lys Trp Tyr Pro Ile Asn Arg Glu Thr Tyr Leu
Asp Arg Leu65 70 75
80Ala Leu Arg Tyr Asp Arg Glu Gly Glu Pro Ser Gln Leu Ala Pro Ile
85 90 95Asp Val Phe Val Ser Thr
Val Asp Pro Leu Lys Glu Pro Pro Leu Ile 100
105 110Thr Gly Asn Thr Val Leu Ser Ile Leu Ala Val Asp
Tyr Pro Val Asp 115 120 125Lys Val
Ser Cys Tyr Val Ser Asp Asp Gly Ser Ala Met Leu Thr Phe 130
135 140Glu Ala Leu Ser Glu Thr Ala Glu Phe Ala Arg
Lys Trp Val Pro Phe145 150 155
160Cys Lys Lys His Asn Ile Glu Pro Arg Ala Pro Glu Phe Tyr Phe Ala
165 170 175Arg Lys Ile Asp
Tyr Leu Lys Asp Lys Ile Gln Pro Ser Phe Val Lys 180
185 190Glu Arg Arg Ala Met Lys Arg Glu Cys Glu Glu
Phe Lys Val Arg Ile 195 200 205Asp
Ala Leu Val Ala Lys Ala Gln Lys Ile Pro Glu Glu Gly Trp Thr 210
215 220Met Ala Asp Gly Thr Pro Trp Pro Gly Asn
Asn Pro Arg Asp His Pro225 230 235
240Gly Met Ile Gln Val Phe Leu Gly His Ser Gly Gly Leu Asp Thr
Asp 245 250 255Gly Asn Glu
Leu Pro Arg Leu Val Tyr Val Ser Arg Glu Lys Arg Pro 260
265 270Gly Phe Gln His His Lys Lys Ala Gly Ala
Met Asn Ala Leu Ile Arg 275 280
285Val Ser Ala Val Leu Thr Asn Gly Ala Tyr Leu Leu Asn Val Asp Cys 290
295 300Asp His Tyr Phe Asn Ser Ser Lys
Ala Leu Arg Glu Ala Met Cys Phe305 310
315 320Met Met Asp Pro Ala Leu Gly Arg Lys Thr Cys Tyr
Val Gln Phe Pro 325 330
335Gln Arg Phe Asp Gly Ile Asp Leu His Asp Arg Tyr Ala Asn Arg Asn
340 345 350Ile Val Phe Phe Asp Ile
Asn Met Lys Gly Leu Asp Gly Ile Gln Gly 355 360
365Pro Val Tyr Val Gly Thr Gly Cys Cys Phe Asn Arg Gln Ala
Leu Tyr 370 375 380Gly Tyr Asp Pro Val
Leu Thr Glu Ala Asp Leu Glu Pro Asn Ile Ile385 390
395 400Ile Lys Ser Cys Cys Gly Gly Arg Lys Lys
Lys Asp Lys Ser Tyr Ile 405 410
415Asp Ser Lys Asn Arg Asp Met Lys Arg Thr Glu Ser Ser Ala Pro Ile
420 425 430Phe Asn Met Glu Asp
Ile Glu Glu Gly Phe Glu Gly Tyr Glu Asp Glu 435
440 445Arg Ser Leu Leu Met Ser Gln Lys Ser Leu Glu Lys
Arg Phe Gly Gln 450 455 460Ser Pro Ile
Phe Ile Ala Ser Thr Phe Met Thr Gln Gly Gly Ile Pro465
470 475 480Pro Ser Thr Asn Pro Gly Ser
Leu Leu Lys Glu Ala Ile His Val Ile 485
490 495Ser Cys Gly Tyr Glu Asp Lys Thr Glu Trp Gly Lys
Glu Ile Gly Trp 500 505 510Ile
Tyr Gly Ser Val Thr Glu Asp Ile Leu Thr Gly Phe Lys Met His 515
520 525Ala Arg Gly Trp Ile Ser Ile Tyr Cys
Met Pro Leu Arg Pro Cys Phe 530 535
540Lys Gly Ser Ala Pro Ile Asn Leu Ser Asp Arg Leu Asn Gln Val Leu545
550 555 560Arg Trp Ala Leu
Gly Ser Val Glu Ile Leu Leu Ser Arg His Cys Pro 565
570 575Ile Trp Tyr Gly Tyr Asn Gly Arg Leu Lys
Leu Leu Glu Arg Leu Ala 580 585
590Tyr Ile Asn Thr Ile Val Tyr Pro Ile Thr Ser Ile Pro Leu Val Ala
595 600 605Tyr Cys Val Leu Pro Ala Ile
Cys Leu Leu Thr Asn Lys Phe Ile Ile 610 615
620Pro Ala Ile Ser Asn Tyr Ala Gly Ala Phe Phe Ile Leu Leu Phe
Ala625 630 635 640Ser Ile
Phe Ala Thr Gly Ile Leu Glu Leu Arg Trp Ser Gly Val Gly
645 650 655Ile Glu Asp Trp Trp Arg Asn
Glu Gln Phe Trp Val Ile Gly Gly Thr 660 665
670Ser Ala His Leu Phe Ala Val Phe Gln Gly Leu Leu Lys Val
Leu Ala 675 680 685Gly Ile Asp Thr
Asn Phe Thr Val Thr Ser Lys Ala Thr Asp Asp Asp 690
695 700Gly Asp Phe Ala Glu Leu Tyr Val Phe Lys Trp Thr
Thr Leu Leu Ile705 710 715
720Pro Pro Thr Thr Val Leu Val Ile Asn Leu Val Gly Ile Val Ala Gly
725 730 735Val Ser Tyr Ala Ile
Asn Ser Gly Tyr Gln Ser Trp Gly Pro Leu Phe 740
745 750Gly Lys Leu Phe Phe Ala Ile Trp Val Ile Leu His
Leu Tyr Pro Phe 755 760 765Leu Lys
Gly Leu Met Gly Lys Gln Asn Arg Thr Pro Thr Ile Val Ile 770
775 780Val Trp Ser Val Leu Leu Ala Ser Ile Phe Ser
Leu Leu Trp Val Lys785 790 795
800Ile Asp Pro Phe Ile Ser Pro Thr Gln Lys Ala Leu Ser Arg Gly Gln
805 810 815Cys Gly Val Asn
Cys 820253773DNAZea mays 25gtcgacccac gcgtccgcta ggatcaaaac
cgtctcgccg ctgcaataat cttttgtcaa 60ttcttaatcc ctcgcgtcga cagcgacagc
ggaaccaact cacgttgccg cggcttcctc 120catcggtgcg gtgccctgtc cttttctctc
gtccctcctc cccccgtata gttaagcccc 180gccccgctac tactactact agcagcagca
gcgctctcgc agcgggagat gcggtgttga 240tccgtgcccc gctcggatct cgggactggt
gccggctctg cccaggcccc aggctccagg 300ccagctccct cgacgtttct cggcgagctc
gcttgccatg gagggcgacg cggacggcgt 360gaagtcgggg aggcgcggtg gcggacaggt
gtgccagatc tgcggcgacg gcgtgggcac 420cacggcggag ggggacgtct tcgccgcctg
cgacgtctgc gggtttccgg tgtgccgccc 480ctgctacgag tacgagcgca aggacggcac
gcaggcgtgc ccccagtgca agaccaagta 540caagcgccac aaggggagcc cggcgatccg
tggggaggaa ggagacgaca ctgatgccga 600tagcgacttc aattaccttg catctggcaa
tgaggaccag aagcagaaga ttgccgacag 660aatgcgcagc tggcgcatga acgttggggg
cagcggggat gttggtcgcc ccaagtatga 720cagtggcgag atcgggctta ccaagtatga
cagtggcgag attcctcggg gatacatccc 780atcagtcact aacagccaga tctcaggaga
aatccctggt gcttcccctg accatcatat 840gatgtcccca actgggaaca ttggcaagcg
tgctccattt ccctatgtga accattcgcc 900aaatccgtca agggagttct ctggtagcat
tgggaatgtt gcctggaaag agagggttga 960tggctggaaa atgaagcagg acaaggggac
gattcccatg acgaatggca caagcattgc 1020tccctctgag ggtcggggtg ttggtgatat
tgatgcatca actgattaca acatggaaga 1080tgccttattg aacgacgaaa ctcgacagcc
tctatctagg aaagttccac ttccttcctc 1140caggataaat ccatacagga tggtcattgt
gctgcgattg attgttctaa gcatcttctt 1200gcactaccgt atcacaaatc ctgtgcgcaa
tgcataccca ttatggcttc tatctgttat 1260atgtgagatc tggtttgctc tttcgtggat
attggatcag ttccctaagt ggtttccaat 1320caaccgggag acgtaccttg ataggctggc
attaaggtat gaccgggaag gtgagccatc 1380tcagttggct gctgttgaca ttttcgtcag
tacagtcgac ccaatgaagg agcctcctct 1440tgtcactgcc aataccgtgc tatccattct
tgctgtggat taccctgtgg ataaggtctc 1500ttgctatgta tctgatgatg gagctgcgat
gctgacattt gatgcactag ctgagacttc 1560agagtttgct agaaaatggg taccatttgt
taagaagtac aacattgaac ctagagctcc 1620tgaatggtac ttctcccaga aaattgatta
cttgaaggac aaagtgcacc cttcatttgt 1680taaagaccgc cgggccatga agagagaata
tgaagaattc aaagttaggg taaatggcct 1740tgttgctaag gcacagaaag ttcctgagga
aggatggatc atgcaagatg gcacaccatg 1800gccaggaaac aataccaggg accatcctgg
aatgattcag gttttccttg gtcacagtgg 1860tggccttgat actgagggca atgagctacc
ccgtttggtc tatgtttctc gtgaaaagcg 1920tcctggattc cagcatcaca agaaagctgg
tgccatgaat gctcttgttc gtgtctcagc 1980tgtgcttacc aatggacaat acatgttgaa
tcttgattgt gatcactaca ttaacaacag 2040taaggctctc agggaagcta tgtgcttcct
tatggaccct aacctaggaa ggagtgtctg 2100ctacgtccag tttccccaga gattcgatgg
cattgacagg aatgatcgat atgccaacag 2160gaacaccgtg tttttcgata ttaacttgag
aggtcttgat ggcatccaag gaccagttta 2220tgtcggaact ggctgtgttt tcaaccgaac
agctctatat ggttatgagc ccccaattaa 2280gcagaagaag ggtggtttct tgtcatcact
atgtggcggt aggaagaagg caagcaaatc 2340aaagaagggc tcggacaaga agaagtcgca
gaagcatgtg gacagttctg tgccagtatt 2400caaccttgaa gatatagagg agggagttga
aggcgctgga tttgacgacg agaaatcact 2460tcttatgtct caaatgagcc tggagaagag
atttggccag tccgcagcgt ttgttgcctc 2520cactctgatg gagtatggtg gtgttcctca
gtccgcaact ccggagtctc ttctgaaaga 2580agctatccat gttataagct gtggctatga
ggacaagact gaatggggaa ctgagatcgg 2640gtggatctac ggttctgtga cagaagacat
tctcaccgga ttcaagatgc acgcgcgagg 2700ctggcggtcg atctactgca tgcccaagcg
gccagctttc aaggggtctg cccccatcaa 2760tctttcggac cgtctgaacc aggtgctccg
gtgggctctt gggtccgtgg agatcctctt 2820cagccggcac tgccccctgt ggtacggcta
cggagggcgg ctcaagttcc tggagagatt 2880cgcgtacatc aacaccacca tctacccgct
cacgtccatc ccgcttctca tctactgcat 2940cctgcccgcc atctgtctgc tcaccggaaa
gttcatcatt ccagagatca gcaacttcgc 3000cagcatctgg ttcatctccc tcttcatctc
gatcttcgcc acgggcatcc tggagatgag 3060gtggagcggg gtgggcatcg acgagtggtg
gaggaacgag cagttctggg tgatcggggg 3120catctccgcg cacctcttcg ccgtgttcca
gggcctgctc aaggtgctgg ccggcatcga 3180caccaacttc accgtcacct ccaaggcctc
ggacgaggac ggcgacttcg cggagctgta 3240catgttcaag tggacgacgc tcctgatccc
gcccaccacc atcctgatca tcaacctggt 3300cggcgtcgtc gccggcatct cctacgccat
caacagcgga taccagtcgt ggggcccgct 3360cttcggcaag ctcttcttcg ccttctgggt
catcgtccac ctgtacccgt tcctcaaggg 3420cctcatgggc aggcagaacc gcaccccgac
catcgtcgtc gtctgggcca tcctgctggc 3480gtccatcttc tccttgctgt gggttcgcat
cgaccccttc accacccgcg tcactggccc 3540ggatacccag acgtgtggca tcaactgcta
gggaagtgga aggtttgtac tttgtagaaa 3600cggaggaata ccacgtgcca tctgttgtct
gttaagttat atatatataa gcagcaagtg 3660gcgttattta cagctacgta cagaccagtg
gatattgttt accacaaagt tttacttgtg 3720ttaatatgca ttcttttgtt gatataaaaa
aaaaaaaaaa aaagggcggc cgc 3773261077PRTZea mays 26Met Glu Gly
Asp Ala Asp Gly Val Lys Ser Gly Arg Arg Gly Gly Gly1 5
10 15Gln Val Cys Gln Ile Cys Gly Asp Gly
Val Gly Thr Thr Ala Glu Gly 20 25
30Asp Val Phe Ala Ala Cys Asp Val Cys Gly Phe Pro Val Cys Arg Pro
35 40 45Cys Tyr Glu Tyr Glu Arg Lys
Asp Gly Thr Gln Ala Cys Pro Gln Cys 50 55
60Lys Thr Lys Tyr Lys Arg His Lys Gly Ser Pro Ala Ile Arg Gly Glu65
70 75 80Glu Gly Asp Asp
Thr Asp Ala Asp Ser Asp Phe Asn Tyr Leu Ala Ser 85
90 95Gly Asn Glu Asp Gln Lys Gln Lys Ile Ala
Asp Arg Met Arg Ser Trp 100 105
110Arg Met Asn Val Gly Gly Ser Gly Asp Val Gly Arg Pro Lys Tyr Asp
115 120 125Ser Gly Glu Ile Gly Leu Thr
Lys Tyr Asp Ser Gly Glu Ile Pro Arg 130 135
140Gly Tyr Ile Pro Ser Val Thr Asn Ser Gln Ile Ser Gly Glu Ile
Pro145 150 155 160Gly Ala
Ser Pro Asp His His Met Met Ser Pro Thr Gly Asn Ile Gly
165 170 175Lys Arg Ala Pro Phe Pro Tyr
Val Asn His Ser Pro Asn Pro Ser Arg 180 185
190Glu Phe Ser Gly Ser Ile Gly Asn Val Ala Trp Lys Glu Arg
Val Asp 195 200 205Gly Trp Lys Met
Lys Gln Asp Lys Gly Thr Ile Pro Met Thr Asn Gly 210
215 220Thr Ser Ile Ala Pro Ser Glu Gly Arg Gly Val Gly
Asp Ile Asp Ala225 230 235
240Ser Thr Asp Tyr Asn Met Glu Asp Ala Leu Leu Asn Asp Glu Thr Arg
245 250 255Gln Pro Leu Ser Arg
Lys Val Pro Leu Pro Ser Ser Arg Ile Asn Pro 260
265 270Tyr Arg Met Val Ile Val Leu Arg Leu Ile Val Leu
Ser Ile Phe Leu 275 280 285His Tyr
Arg Ile Thr Asn Pro Val Arg Asn Ala Tyr Pro Leu Trp Leu 290
295 300Leu Ser Val Ile Cys Glu Ile Trp Phe Ala Leu
Ser Trp Ile Leu Asp305 310 315
320Gln Phe Pro Lys Trp Phe Pro Ile Asn Arg Glu Thr Tyr Leu Asp Arg
325 330 335Leu Ala Leu Arg
Tyr Asp Arg Glu Gly Glu Pro Ser Gln Leu Ala Ala 340
345 350Val Asp Ile Phe Val Ser Thr Val Asp Pro Met
Lys Glu Pro Pro Leu 355 360 365Val
Thr Ala Asn Thr Val Leu Ser Ile Leu Ala Val Asp Tyr Pro Val 370
375 380Asp Lys Val Ser Cys Tyr Val Ser Asp Asp
Gly Ala Ala Met Leu Thr385 390 395
400Phe Asp Ala Leu Ala Glu Thr Ser Glu Phe Ala Arg Lys Trp Val
Pro 405 410 415Phe Val Lys
Lys Tyr Asn Ile Glu Pro Arg Ala Pro Glu Trp Tyr Phe 420
425 430Ser Gln Lys Ile Asp Tyr Leu Lys Asp Lys
Val His Pro Ser Phe Val 435 440
445Lys Asp Arg Arg Ala Met Lys Arg Glu Tyr Glu Glu Phe Lys Val Arg 450
455 460Val Asn Gly Leu Val Ala Lys Ala
Gln Lys Val Pro Glu Glu Gly Trp465 470
475 480Ile Met Gln Asp Gly Thr Pro Trp Pro Gly Asn Asn
Thr Arg Asp His 485 490
495Pro Gly Met Ile Gln Val Phe Leu Gly His Ser Gly Gly Leu Asp Thr
500 505 510Glu Gly Asn Glu Leu Pro
Arg Leu Val Tyr Val Ser Arg Glu Lys Arg 515 520
525Pro Gly Phe Gln His His Lys Lys Ala Gly Ala Met Asn Ala
Leu Val 530 535 540Arg Val Ser Ala Val
Leu Thr Asn Gly Gln Tyr Met Leu Asn Leu Asp545 550
555 560Cys Asp His Tyr Ile Asn Asn Ser Lys Ala
Leu Arg Glu Ala Met Cys 565 570
575Phe Leu Met Asp Pro Asn Leu Gly Arg Ser Val Cys Tyr Val Gln Phe
580 585 590Pro Gln Arg Phe Asp
Gly Ile Asp Arg Asn Asp Arg Tyr Ala Asn Arg 595
600 605Asn Thr Val Phe Phe Asp Ile Asn Leu Arg Gly Leu
Asp Gly Ile Gln 610 615 620Gly Pro Val
Tyr Val Gly Thr Gly Cys Val Phe Asn Arg Thr Ala Leu625
630 635 640Tyr Gly Tyr Glu Pro Pro Ile
Lys Gln Lys Lys Gly Gly Phe Leu Ser 645
650 655Ser Leu Cys Gly Gly Arg Lys Lys Ala Ser Lys Ser
Lys Lys Gly Ser 660 665 670Asp
Lys Lys Lys Ser Gln Lys His Val Asp Ser Ser Val Pro Val Phe 675
680 685Asn Leu Glu Asp Ile Glu Glu Gly Val
Glu Gly Ala Gly Phe Asp Asp 690 695
700Glu Lys Ser Leu Leu Met Ser Gln Met Ser Leu Glu Lys Arg Phe Gly705
710 715 720Gln Ser Ala Ala
Phe Val Ala Ser Thr Leu Met Glu Tyr Gly Gly Val 725
730 735Pro Gln Ser Ala Thr Pro Glu Ser Leu Leu
Lys Glu Ala Ile His Val 740 745
750Ile Ser Cys Gly Tyr Glu Asp Lys Thr Glu Trp Gly Thr Glu Ile Gly
755 760 765Trp Ile Tyr Gly Ser Val Thr
Glu Asp Ile Leu Thr Gly Phe Lys Met 770 775
780His Ala Arg Gly Trp Arg Ser Ile Tyr Cys Met Pro Lys Arg Pro
Ala785 790 795 800Phe Lys
Gly Ser Ala Pro Ile Asn Leu Ser Asp Arg Leu Asn Gln Val
805 810 815Leu Arg Trp Ala Leu Gly Ser
Val Glu Ile Leu Phe Ser Arg His Cys 820 825
830Pro Leu Trp Tyr Gly Tyr Gly Gly Arg Leu Lys Phe Leu Glu
Arg Phe 835 840 845Ala Tyr Ile Asn
Thr Thr Ile Tyr Pro Leu Thr Ser Ile Pro Leu Leu 850
855 860Ile Tyr Cys Ile Leu Pro Ala Ile Cys Leu Leu Thr
Gly Lys Phe Ile865 870 875
880Ile Pro Glu Ile Ser Asn Phe Ala Ser Ile Trp Phe Ile Ser Leu Phe
885 890 895Ile Ser Ile Phe Ala
Thr Gly Ile Leu Glu Met Arg Trp Ser Gly Val 900
905 910Gly Ile Asp Glu Trp Trp Arg Asn Glu Gln Phe Trp
Val Ile Gly Gly 915 920 925Ile Ser
Ala His Leu Phe Ala Val Phe Gln Gly Leu Leu Lys Val Leu 930
935 940Ala Gly Ile Asp Thr Asn Phe Thr Val Thr Ser
Lys Ala Ser Asp Glu945 950 955
960Asp Gly Asp Phe Ala Glu Leu Tyr Met Phe Lys Trp Thr Thr Leu Leu
965 970 975Ile Pro Pro Thr
Thr Ile Leu Ile Ile Asn Leu Val Gly Val Val Ala 980
985 990Gly Ile Ser Tyr Ala Ile Asn Ser Gly Tyr Gln
Ser Trp Gly Pro Leu 995 1000
1005Phe Gly Lys Leu Phe Phe Ala Phe Trp Val Ile Val His Leu Tyr
1010 1015 1020Pro Phe Leu Lys Gly Leu
Met Gly Arg Gln Asn Arg Thr Pro Thr 1025 1030
1035Ile Val Val Val Trp Ala Ile Leu Leu Ala Ser Ile Phe Ser
Leu 1040 1045 1050Leu Trp Val Arg Ile
Asp Pro Phe Thr Thr Arg Val Thr Gly Pro 1055 1060
1065Asp Thr Gln Thr Cys Gly Ile Asn Cys 1070
1075273704DNAZea mays 27gtcgacccac gcttccggtc ggttccgcgt cccttttccc
ctcccccctc cgtcgccgcc 60tcgagcgagc tccaccactt gctcctgcgc gaggtgaaca
ctgggttagg gccactgcca 120ccgctgggct gcctctgctt ctgcctctcc cgccagcgcg
cgagcccggg ggcgattcgg 180cgccggcacg cgggagggga agccgaggaa tgcggtgagt
cggcgggggt ccggcgtttg 240tgaactcgtg gagggctcgg attggtgcgc catggacggc
ggcgacgcca cgaattcggg 300gaagcatgtg gccgggcagg tgtgccagat ctgcggcgac
ggcgtgggca ccgcggcgga 360cggcgacctc ttcaccgcct gcgacgtctg cggcttcccc
gtgtgccgcc catgctacga 420gtacgagcgc aaggacggca cccaggcgtg cccgcagtgc
aagactaagt acaagcgcca 480caaagggagc ccaccagtac acggtgagga aaatgaggat
gtggatgctg acgatgtgag 540tgactacaac taccaagcat ctggcaacca ggatcagaag
caaaagattg ctgagagaat 600gctcacttgg cggacaaact cacgtggcag tgatattggc
ctggctaagt atgacagcgg 660tgaaattggg catgggaagt atgacagtgg tgagatccct
cgtggatata tcccgtcact 720aactcatagc cagatctcag gagagattcc tggagcttcc
cctgatcata tgatgtctcc 780tgttgggaac attggcaggc gtggacatca atttccttat
gtaaatcatt ctccaaaccc 840atcgagggag ttctccggta gccttggcaa tgttgcatgg
aaagagaggg tggatggatg 900gaaaatgaag gataaaggtg caattcctat gaccaatgga
acaagcattg ctccatcaga 960agggcgtgga gttgctgata ttgatgcttc tactgattat
aacatggaag atgccttact 1020gaatgatgaa actcggcaac ctctatctag aaaagtgcca
attccttcat ccagaataaa 1080tccgtacaga atggtcattg tgctacgttt ggctgttcta
tgcatattct tgcgctaccg 1140tatcacacat cctgtgaaca atgcatatcc actgtggctt
ttatccgtca tatgtgagat 1200ctggtttgct ttgtcctgga ttttggatca gttcccaaag
tggtccccaa tcaaccgtga 1260aacatacctt gatagactgg ctttaaggta tgaccgagaa
ggtgaaccat ctcaattagc 1320tcctgttgat atttttgtca gtactgtgga tccaatgaag
gagcctcctc ttgtcactgc 1380aaatactgtg ctttccatcc ttgctgtcga ttatccggtt
gacaaggtat cttgctatgt 1440ttcggatgat ggagctgcta tgctgacttt tgatgctctc
tctgaaactt cagagtttgc 1500tagaaaatgg gttccgttct gtaagaagta caacatagag
cctagggccc cggaatggta 1560ctttgctcag aaaattgatt acttgaaaga caaagttcaa
acctcatttg tgaaagaacg 1620ccgggccatg aagagagaat atgaagaatt caaagttcgt
atcaatggtc ttgtagccaa 1680ggcacaaaaa gttcccgagg agggatggat catgcaagat
ggtacacctt ggcctgggaa 1740caatactagg gaccatcctg gaatgattca ggttttcctg
ggtcacagtg gagggcttga 1800cgttgaaggc aatgaacttc ctcgtttggt ttatgtgtct
cgtgaaaaac gtcctggatt 1860ccaacatcac aagaaggctg gtgccatgaa tgcacttgtt
cgtgtatcag ctgtccttac 1920taatgggcaa tacatgttga atcttgattg tgaccactac
atcaataata gcaaggctct 1980tcgagaagct atgtgcttcc ttatggaccc aaacctagga
aggaatgtct gttatgtcca 2040atttcctcag aggtttgatg gtattgatag gaatgaccga
tatgcaaaca ggaacactgt 2100gtttttcgat attaacttga gaggtcttga cggcattcaa
gggccagttt atgtgggaac 2160tggttgtgtg tttaacagaa cggccttata tggttatgag
cctccagtca agaaaaaaaa 2220gccaggcttc ttctcttcgc tttgtggggg aaggaaaaag
acgtcaaaat ctaagaagag 2280ctcggaaaag aagaagtcac atagacacgc agacagttct
gtaccagtat ttaatctcga 2340agatatagag gaagggattg aaggttctca gtttgatgat
gagaaatcgc tgattatgtc 2400tcaaatgagc ttggagaaga gatttggcca gtccagtgtt
tttgtagcct ctactctgat 2460ggaatatggt ggtgttccac aatctgcaac tccagagtct
cttctgaaag aagctattca 2520tgtcatcagc tgtggctatg aggacaaaac tgactgggga
actgagattg ggtggatcta 2580tggttctgtt acagaagaca ttctcaccgg attcaagatg
catgctcgag gctggcgatc 2640aatctactgc atgcctaagc gaccagcttt caagggatct
gctcctatca acctttcgga 2700tcgtttgaat caagtgcttc ggtgggctct tggttccatt
gaaattcttt tcagcaggca 2760ttgtcccata tggtatggct atggaggccg gcttaaattc
ctggagagat ttgcttatat 2820caacacaaca atttatccac tcacatcaat cccgctcctc
ctgtactgca tattgccagc 2880agtttgtctt ctcactggga agttcatcat cccaaagatt
agtaacctag agagtgtttg 2940gtttatatcg ctctttatct caatctttgc cactggtatc
cttgagatga ggtggagtgg 3000tgttggcatt gatgaatggt ggaggaacga gcagttctgg
gtcattggtg gtatttctgc 3060gcatttattt gccgtcttcc agggtctcct gaaggtgctt
gctggtatcg acacgagctt 3120cactgtcacc tctaaggcca ctgacgaaga aggtgatttt
gccgagctct acatgttcaa 3180gtggacaacg cttctgatcc caccaaccac tattttgatc
atcaacctgg tcggcgtggt 3240cgctggcatt tcctacgcaa tcaatagcgg ttaccagtca
tggggacctc ttttcgggaa 3300gctcttcttt gcgttctggg tgattgtcca cctgtacccc
ttcctcaagg gcctcatggg 3360gaagcagaac cgcacgccga ccattgtcgt tgtctgggct
atcctccttg cgtcgatctt 3420ttccctgatg tgggttcgta tcgatccatt caccacccgg
gtcactggcc ctgatatcgc 3480gaaatgtggc atcaactgct aggatgagct gaagatagtt
aaagagtgga actagacgca 3540ttgtgcatcg taagttatca gtgggtggct ctttttatag
tatggtagga acttggtcgg 3600gagacgttaa ttacatatgc tatatgtacc tccgctggtc
tttatccgta agttaatata 3660tatactgctt tgagaattaa aaaaaaaaaa aaaagggcgg
ccgc 3704281076PRTZea mays 28Met Asp Gly Gly Asp Ala
Thr Asn Ser Gly Lys His Val Ala Gly Gln1 5
10 15Val Cys Gln Ile Cys Gly Asp Gly Val Gly Thr Ala
Ala Asp Gly Asp 20 25 30Leu
Phe Thr Ala Cys Asp Val Cys Gly Phe Pro Val Cys Arg Pro Cys 35
40 45Tyr Glu Tyr Glu Arg Lys Asp Gly Thr
Gln Ala Cys Pro Gln Cys Lys 50 55
60Thr Lys Tyr Lys Arg His Lys Gly Ser Pro Pro Val His Gly Glu Glu65
70 75 80Asn Glu Asp Val Asp
Ala Asp Asp Val Ser Asp Tyr Asn Tyr Gln Ala 85
90 95Ser Gly Asn Gln Asp Gln Lys Gln Lys Ile Ala
Glu Arg Met Leu Thr 100 105
110Trp Arg Thr Asn Ser Arg Gly Ser Asp Ile Gly Leu Ala Lys Tyr Asp
115 120 125Ser Gly Glu Ile Gly His Gly
Lys Tyr Asp Ser Gly Glu Ile Pro Arg 130 135
140Gly Tyr Ile Pro Ser Leu Thr His Ser Gln Ile Ser Gly Glu Ile
Pro145 150 155 160Gly Ala
Ser Pro Asp His Met Met Ser Pro Val Gly Asn Ile Gly Arg
165 170 175Arg Gly His Gln Phe Pro Tyr
Val Asn His Ser Pro Asn Pro Ser Arg 180 185
190Glu Phe Ser Gly Ser Leu Gly Asn Val Ala Trp Lys Glu Arg
Val Asp 195 200 205Gly Trp Lys Met
Lys Asp Lys Gly Ala Ile Pro Met Thr Asn Gly Thr 210
215 220Ser Ile Ala Pro Ser Glu Gly Arg Gly Val Ala Asp
Ile Asp Ala Ser225 230 235
240Thr Asp Tyr Asn Met Glu Asp Ala Leu Leu Asn Asp Glu Thr Arg Gln
245 250 255Pro Leu Ser Arg Lys
Val Pro Ile Pro Ser Ser Arg Ile Asn Pro Tyr 260
265 270Arg Met Val Ile Val Leu Arg Leu Ala Val Leu Cys
Ile Phe Leu Arg 275 280 285Tyr Arg
Ile Thr His Pro Val Asn Asn Ala Tyr Pro Leu Trp Leu Leu 290
295 300Ser Val Ile Cys Glu Ile Trp Phe Ala Leu Ser
Trp Ile Leu Asp Gln305 310 315
320Phe Pro Lys Trp Ser Pro Ile Asn Arg Glu Thr Tyr Leu Asp Arg Leu
325 330 335Ala Leu Arg Tyr
Asp Arg Glu Gly Glu Pro Ser Gln Leu Ala Pro Val 340
345 350Asp Ile Phe Val Ser Thr Val Asp Pro Met Lys
Glu Pro Pro Leu Val 355 360 365Thr
Ala Asn Thr Val Leu Ser Ile Leu Ala Val Asp Tyr Pro Val Asp 370
375 380Lys Val Ser Cys Tyr Val Ser Asp Asp Gly
Ala Ala Met Leu Thr Phe385 390 395
400Asp Ala Leu Ser Glu Thr Ser Glu Phe Ala Arg Lys Trp Val Pro
Phe 405 410 415Cys Lys Lys
Tyr Asn Ile Glu Pro Arg Ala Pro Glu Trp Tyr Phe Ala 420
425 430Gln Lys Ile Asp Tyr Leu Lys Asp Lys Val
Gln Thr Ser Phe Val Lys 435 440
445Glu Arg Arg Ala Met Lys Arg Glu Tyr Glu Glu Phe Lys Val Arg Ile 450
455 460Asn Gly Leu Val Ala Lys Ala Gln
Lys Val Pro Glu Glu Gly Trp Ile465 470
475 480Met Gln Asp Gly Thr Pro Trp Pro Gly Asn Asn Thr
Arg Asp His Pro 485 490
495Gly Met Ile Gln Val Phe Leu Gly His Ser Gly Gly Leu Asp Val Glu
500 505 510Gly Asn Glu Leu Pro Arg
Leu Val Tyr Val Ser Arg Glu Lys Arg Pro 515 520
525Gly Phe Gln His His Lys Lys Ala Gly Ala Met Asn Ala Leu
Val Arg 530 535 540Val Ser Ala Val Leu
Thr Asn Gly Gln Tyr Met Leu Asn Leu Asp Cys545 550
555 560Asp His Tyr Ile Asn Asn Ser Lys Ala Leu
Arg Glu Ala Met Cys Phe 565 570
575Leu Met Asp Pro Asn Leu Gly Arg Asn Val Cys Tyr Val Gln Phe Pro
580 585 590Gln Arg Phe Asp Gly
Ile Asp Arg Asn Asp Arg Tyr Ala Asn Arg Asn 595
600 605Thr Val Phe Phe Asp Ile Asn Leu Arg Gly Leu Asp
Gly Ile Gln Gly 610 615 620Pro Val Tyr
Val Gly Thr Gly Cys Val Phe Asn Arg Thr Ala Leu Tyr625
630 635 640Gly Tyr Glu Pro Pro Val Lys
Lys Lys Lys Pro Gly Phe Phe Ser Ser 645
650 655Leu Cys Gly Gly Arg Lys Lys Thr Ser Lys Ser Lys
Lys Ser Ser Glu 660 665 670Lys
Lys Lys Ser His Arg His Ala Asp Ser Ser Val Pro Val Phe Asn 675
680 685Leu Glu Asp Ile Glu Glu Gly Ile Glu
Gly Ser Gln Phe Asp Asp Glu 690 695
700Lys Ser Leu Ile Met Ser Gln Met Ser Leu Glu Lys Arg Phe Gly Gln705
710 715 720Ser Ser Val Phe
Val Ala Ser Thr Leu Met Glu Tyr Gly Gly Val Pro 725
730 735Gln Ser Ala Thr Pro Glu Ser Leu Leu Lys
Glu Ala Ile His Val Ile 740 745
750Ser Cys Gly Tyr Glu Asp Lys Thr Asp Trp Gly Thr Glu Ile Gly Trp
755 760 765Ile Tyr Gly Ser Val Thr Glu
Asp Ile Leu Thr Gly Phe Lys Met His 770 775
780Ala Arg Gly Trp Arg Ser Ile Tyr Cys Met Pro Lys Arg Pro Ala
Phe785 790 795 800Lys Gly
Ser Ala Pro Ile Asn Leu Ser Asp Arg Leu Asn Gln Val Leu
805 810 815Arg Trp Ala Leu Gly Ser Ile
Glu Ile Leu Phe Ser Arg His Cys Pro 820 825
830Ile Trp Tyr Gly Tyr Gly Gly Arg Leu Lys Phe Leu Glu Arg
Phe Ala 835 840 845Tyr Ile Asn Thr
Thr Ile Tyr Pro Leu Thr Ser Ile Pro Leu Leu Leu 850
855 860Tyr Cys Ile Leu Pro Ala Val Cys Leu Leu Thr Gly
Lys Phe Ile Ile865 870 875
880Pro Lys Ile Ser Asn Leu Glu Ser Val Trp Phe Ile Ser Leu Phe Ile
885 890 895Ser Ile Phe Ala Thr
Gly Ile Leu Glu Met Arg Trp Ser Gly Val Gly 900
905 910Ile Asp Glu Trp Trp Arg Asn Glu Gln Phe Trp Val
Ile Gly Gly Ile 915 920 925Ser Ala
His Leu Phe Ala Val Phe Gln Gly Leu Leu Lys Val Leu Ala 930
935 940Gly Ile Asp Thr Ser Phe Thr Val Thr Ser Lys
Ala Thr Asp Glu Glu945 950 955
960Gly Asp Phe Ala Glu Leu Tyr Met Phe Lys Trp Thr Thr Leu Leu Ile
965 970 975Pro Pro Thr Thr
Ile Leu Ile Ile Asn Leu Val Gly Val Val Ala Gly 980
985 990Ile Ser Tyr Ala Ile Asn Ser Gly Tyr Gln Ser
Trp Gly Pro Leu Phe 995 1000
1005Gly Lys Leu Phe Phe Ala Phe Trp Val Ile Val His Leu Tyr Pro
1010 1015 1020Phe Leu Lys Gly Leu Met
Gly Lys Gln Asn Arg Thr Pro Thr Ile 1025 1030
1035Val Val Val Trp Ala Ile Leu Leu Ala Ser Ile Phe Ser Leu
Met 1040 1045 1050Trp Val Arg Ile Asp
Pro Phe Thr Thr Arg Val Thr Gly Pro Asp 1055 1060
1065Ile Ala Lys Cys Gly Ile Asn Cys 1070
1075293568DNAZea maysmisc_feature(3487)..(3487)n is a, c, g, or t
29gtcgacccac gcgtccggag ctcgtcgtca tccgccgcga tggcgagcca gggccgaagc
60ccatggacca gcggaacggc caggtgtgcc agatttgcgg cgacgacgtg gggcgcaacc
120ccgacgggga gcctttcgtg gcctgcaacg agtgcgcctt ccccatctgc cgggactgct
180acgagtacga gcgccgcgag ggcacgcaga actgccccca gtgcaagacc cgcttcaagc
240gcttcaaggg gtgcgcgcgc gtgcccgggg acgaggagga ggacggcgtc gacgacctgg
300agaacgagtt caactggagc gacaagcacg actcccagta cctcgccgag tccatgctcc
360acgcccacat gagctacggc cgcggcgccg acctcgacgg cgtgccgcag ccattccacc
420ccatccccaa tgttcccctc ctcaccaacg gacagatggt cgatgacatc ccgccggacc
480agcacgccct tgtgccctcg ttcgtgggtg gcggggggaa gaggattcac cctctcccgt
540acgcggatcc caaccttcct gtgcaaccga ggtctatgga cccttccaag gatctcgccg
600catatggcta cgggagcgta gcatggaagg agaggatgga gagctggaag cagaagcagg
660agaggatgca ccagacgagg aacgatggcg gcggcgatga tggtgatgat gcagatctac
720cactaatgga tgaagctaga cagccattgt ccagaaagat cccgcttcct tcaagccaaa
780tcaaccccta taggatgatt ataataattc ggctagtggt tttgtgtttc ttcttccact
840accgagtgat gcatccggtg cctgatgcat ttgctttatg gctcatatct gtgatctgtg
900aaatttggtt tgccatgtct tggattcttg accagtttcc aaagtggttt cctatcgaga
960gggaaaccta tcttgaccgg ctgagtttaa ggtttgacaa ggaagggcat ccttctcaac
1020tcgcccctgt tgatttcttt gtcagtacgg ttgatccctt gaaggaacct ccattggtca
1080ctgctaatac tgttctatct atcctttcgg tggattatcc agttgataag gtttcatgct
1140acgtttctga tgatggtgct gccatgctga catttgaagc attgtctgaa acatctgaat
1200ttgcaaagaa atgggttcct ttctgcaaaa gatatagcct tgagcctcgt gctccagagt
1260ggtacttcca acagaagata gactacctga aagacaaggt ggcgccaaac tttgttagag
1320aacggagagc aatgaagaga gagtatgagg aattcaaggt cagaatcaat gccttggttg
1380ctaaagccca aaaggttcct gaggaaggat ggacaatgca ggatggaact ccatggcccg
1440gaaataatgt ccgtgatcat cctggaatga ttcaggtttt ccttggtcaa agtggtggcc
1500atgatgtgga aggaaatgag ctgcctcgat tggtttatgt ttcaagagaa aaacggccag
1560gctacaacca tcacaagaag gctggtgcta tgaatgcatt ggtccgagtc tctgctgtac
1620taactaatgc tccttatttg ctgaacttgg attgtgatca ctatatcaat aatagtaagg
1680ctataaagga agcaatgtgt tttatgatgg atcctttgct tggaaagaaa gtttgctatg
1740tgcagtttcc tcaaagattt gatgggattg atcgccatga tcgatatgct aacagaaatg
1800ttgtcttttt cgatatcaac atgaaaggtt tggatggtat ccagggccca atttatgtgg
1860gtactggatg tgtcttcaga aggcaggcat tatatggcta cgatgctccc aaaacaaaga
1920agccaccatc aagaacttgc aactgctggc caaagtggtg catttgctgt tgctgttttg
1980gtaacaggaa gaccaagaag aagaccaaga cctctaaacc taaatttgag aagataaaga
2040aactttttaa gaaaaaggaa aatcaagccc ctgcatatgc tcttggtgaa attgatgaag
2100ccgctccagg agctgaaaat gaaaaggcta gtattgtaaa tcaacagaag ttggaaaaga
2160aatttggcca gtcttcagtt tttgttgcat ccacacttct tgagaatggt ggaaccctga
2220agagtgccag tccagcttct cttctgaagg aagctataca tgtcatcagt tgtggatatg
2280aagacaaaac aggctgggga aaagatattg gttggattta tggatcagtc acagaagata
2340ttcttactgg gtttaagatg cactgccatg gttggcggtc aatttactgc atacctaaac
2400gggccgcctt caaaggttcc gcacctctca atctttccga tcgttttcac caggttcttc
2460ggtgggctct tggttcaatt gaaattttgt tcagcaacca ctgccctctc tggtatgggt
2520atggtggtgg actaaagttc ctggaaaggt tttcgtacat taactccatc gtataccctt
2580ggacatctat cccgctcttg gcctattgca cattgcctgc catctgcttg ctgacaggga
2640aatttatcac gccagagctt aacaatgttg ccagcctctg gttcatgtca cttttcatct
2700gcatttttgc tacgagcatc ctggaaatga gatggagtgg tgtaggcatc gatgactggt
2760ggagaaacga gcagttttgg gtcattggag gcgtgtcttc acatctcttt gctgtgttcc
2820agggactcct caaggtcata gctggtgtag acacgagctt cactgtgaca tccaagggcg
2880gagacgacga ggagttctca gagctgtaca cattcaaatg gacgaccctt ctgatacctc
2940cgacaaccct gctcctactg aacttcattg gagtggtagc tggcatctcc aatgcgatca
3000acaacggata tgaatcatgg ggccccctgt tcgggaagct cttctttgca ttttgggtga
3060tcgtccatct ttacccgttc ctcaagggtc tggttgggag gcagaacagg acgccaacga
3120ttgtcattgt ctggtccatc ctcctggctt cgatcttctc gctgctttgg gtccggatcg
3180acccgttcct tgcgaaggat gatggtcccc tgttggagga gtgtggtctg gattgcaact
3240aggaggtcag cacgtggact tccccgtcag tgtgtggtcg aagaagtatt tttgcagatg
3300ttttgtgccc atatttcttt actcaatttt tgtccctctg tagattgaaa caaggggtga
3360aggggaaaaa aagtacttgt atttcttttg ttccatggtg gtggtggtgg tgggcggctc
3420agcctcgtga gtgcaatatt gggcaaaccg gaggttgcgg caaccttgtg cagttcgtcc
3480acgaatntac tagggatgat cgcgaccaat caatcaatcg atgaccgagt tcaattgttc
3540aaaaaaaaaa aaaaaaaagg gcggccgc
3568301059PRTZea mays 30Met Asp Gln Arg Asn Gly Gln Val Cys Gln Ile Cys
Gly Asp Asp Val1 5 10
15Gly Arg Asn Pro Asp Gly Glu Pro Phe Val Ala Cys Asn Glu Cys Ala
20 25 30Phe Pro Ile Cys Arg Asp Cys
Tyr Glu Tyr Glu Arg Arg Glu Gly Thr 35 40
45Gln Asn Cys Pro Gln Cys Lys Thr Arg Phe Lys Arg Phe Lys Gly
Cys 50 55 60Ala Arg Val Pro Gly Asp
Glu Glu Glu Asp Gly Val Asp Asp Leu Glu65 70
75 80Asn Glu Phe Asn Trp Ser Asp Lys His Asp Ser
Gln Tyr Leu Ala Glu 85 90
95Ser Met Leu His Ala His Met Ser Tyr Gly Arg Gly Ala Asp Leu Asp
100 105 110Gly Val Pro Gln Pro Phe
His Pro Ile Pro Asn Val Pro Leu Leu Thr 115 120
125Asn Gly Gln Met Val Asp Asp Ile Pro Pro Asp Gln His Ala
Leu Val 130 135 140Pro Ser Phe Val Gly
Gly Gly Gly Lys Arg Ile His Pro Leu Pro Tyr145 150
155 160Ala Asp Pro Asn Leu Pro Val Gln Pro Arg
Ser Met Asp Pro Ser Lys 165 170
175Asp Leu Ala Ala Tyr Gly Tyr Gly Ser Val Ala Trp Lys Glu Arg Met
180 185 190Glu Ser Trp Lys Gln
Lys Gln Glu Arg Met His Gln Thr Arg Asn Asp 195
200 205Gly Gly Gly Asp Asp Gly Asp Asp Ala Asp Leu Pro
Leu Met Asp Glu 210 215 220Ala Arg Gln
Pro Leu Ser Arg Lys Ile Pro Leu Pro Ser Ser Gln Ile225
230 235 240Asn Pro Tyr Arg Met Ile Ile
Ile Ile Arg Leu Val Val Leu Cys Phe 245
250 255Phe Phe His Tyr Arg Val Met His Pro Val Pro Asp
Ala Phe Ala Leu 260 265 270Trp
Leu Ile Ser Val Ile Cys Glu Ile Trp Phe Ala Met Ser Trp Ile 275
280 285Leu Asp Gln Phe Pro Lys Trp Phe Pro
Ile Glu Arg Glu Thr Tyr Leu 290 295
300Asp Arg Leu Ser Leu Arg Phe Asp Lys Glu Gly His Pro Ser Gln Leu305
310 315 320Ala Pro Val Asp
Phe Phe Val Ser Thr Val Asp Pro Leu Lys Glu Pro 325
330 335Pro Leu Val Thr Ala Asn Thr Val Leu Ser
Ile Leu Ser Val Asp Tyr 340 345
350Pro Val Asp Lys Val Ser Cys Tyr Val Ser Asp Asp Gly Ala Ala Met
355 360 365Leu Thr Phe Glu Ala Leu Ser
Glu Thr Ser Glu Phe Ala Lys Lys Trp 370 375
380Val Pro Phe Cys Lys Arg Tyr Ser Leu Glu Pro Arg Ala Pro Glu
Trp385 390 395 400Tyr Phe
Gln Gln Lys Ile Asp Tyr Leu Lys Asp Lys Val Ala Pro Asn
405 410 415Phe Val Arg Glu Arg Arg Ala
Met Lys Arg Glu Tyr Glu Glu Phe Lys 420 425
430Val Arg Ile Asn Ala Leu Val Ala Lys Ala Gln Lys Val Pro
Glu Glu 435 440 445Gly Trp Thr Met
Gln Asp Gly Thr Pro Trp Pro Gly Asn Asn Val Arg 450
455 460Asp His Pro Gly Met Ile Gln Val Phe Leu Gly Gln
Ser Gly Gly His465 470 475
480Asp Val Glu Gly Asn Glu Leu Pro Arg Leu Val Tyr Val Ser Arg Glu
485 490 495Lys Arg Pro Gly Tyr
Asn His His Lys Lys Ala Gly Ala Met Asn Ala 500
505 510Leu Val Arg Val Ser Ala Val Leu Thr Asn Ala Pro
Tyr Leu Leu Asn 515 520 525Leu Asp
Cys Asp His Tyr Ile Asn Asn Ser Lys Ala Ile Lys Glu Ala 530
535 540Met Cys Phe Met Met Asp Pro Leu Leu Gly Lys
Lys Val Cys Tyr Val545 550 555
560Gln Phe Pro Gln Arg Phe Asp Gly Ile Asp Arg His Asp Arg Tyr Ala
565 570 575Asn Arg Asn Val
Val Phe Phe Asp Ile Asn Met Lys Gly Leu Asp Gly 580
585 590Ile Gln Gly Pro Ile Tyr Val Gly Thr Gly Cys
Val Phe Arg Arg Gln 595 600 605Ala
Leu Tyr Gly Tyr Asp Ala Pro Lys Thr Lys Lys Pro Pro Ser Arg 610
615 620Thr Cys Asn Cys Trp Pro Lys Trp Cys Ile
Cys Cys Cys Cys Phe Gly625 630 635
640Asn Arg Lys Thr Lys Lys Lys Thr Lys Thr Ser Lys Pro Lys Phe
Glu 645 650 655Lys Ile Lys
Lys Leu Phe Lys Lys Lys Glu Asn Gln Ala Pro Ala Tyr 660
665 670Ala Leu Gly Glu Ile Asp Glu Ala Ala Pro
Gly Ala Glu Asn Glu Lys 675 680
685Ala Ser Ile Val Asn Gln Gln Lys Leu Glu Lys Lys Phe Gly Gln Ser 690
695 700Ser Val Phe Val Ala Ser Thr Leu
Leu Glu Asn Gly Gly Thr Leu Lys705 710
715 720Ser Ala Ser Pro Ala Ser Leu Leu Lys Glu Ala Ile
His Val Ile Ser 725 730
735Cys Gly Tyr Glu Asp Lys Thr Gly Trp Gly Lys Asp Ile Gly Trp Ile
740 745 750Tyr Gly Ser Val Thr Glu
Asp Ile Leu Thr Gly Phe Lys Met His Cys 755 760
765His Gly Trp Arg Ser Ile Tyr Cys Ile Pro Lys Arg Ala Ala
Phe Lys 770 775 780Gly Ser Ala Pro Leu
Asn Leu Ser Asp Arg Phe His Gln Val Leu Arg785 790
795 800Trp Ala Leu Gly Ser Ile Glu Ile Leu Phe
Ser Asn His Cys Pro Leu 805 810
815Trp Tyr Gly Tyr Gly Gly Gly Leu Lys Phe Leu Glu Arg Phe Ser Tyr
820 825 830Ile Asn Ser Ile Val
Tyr Pro Trp Thr Ser Ile Pro Leu Leu Ala Tyr 835
840 845Cys Thr Leu Pro Ala Ile Cys Leu Leu Thr Gly Lys
Phe Ile Thr Pro 850 855 860Glu Leu Asn
Asn Val Ala Ser Leu Trp Phe Met Ser Leu Phe Ile Cys865
870 875 880Ile Phe Ala Thr Ser Ile Leu
Glu Met Arg Trp Ser Gly Val Gly Ile 885
890 895Asp Asp Trp Trp Arg Asn Glu Gln Phe Trp Val Ile
Gly Gly Val Ser 900 905 910Ser
His Leu Phe Ala Val Phe Gln Gly Leu Leu Lys Val Ile Ala Gly 915
920 925Val Asp Thr Ser Phe Thr Val Thr Ser
Lys Gly Gly Asp Asp Glu Glu 930 935
940Phe Ser Glu Leu Tyr Thr Phe Lys Trp Thr Thr Leu Leu Ile Pro Pro945
950 955 960Thr Thr Leu Leu
Leu Leu Asn Phe Ile Gly Val Val Ala Gly Ile Ser 965
970 975Asn Ala Ile Asn Asn Gly Tyr Glu Ser Trp
Gly Pro Leu Phe Gly Lys 980 985
990Leu Phe Phe Ala Phe Trp Val Ile Val His Leu Tyr Pro Phe Leu Lys
995 1000 1005Gly Leu Val Gly Arg Gln
Asn Arg Thr Pro Thr Ile Val Ile Val 1010 1015
1020Trp Ser Ile Leu Leu Ala Ser Ile Phe Ser Leu Leu Trp Val
Arg 1025 1030 1035Ile Asp Pro Phe Leu
Ala Lys Asp Asp Gly Pro Leu Leu Glu Glu 1040 1045
1050Cys Gly Leu Asp Cys Asn 1055313969DNAZea mays
31cttctccctc gtcggtgcgg cgtggcgcgg ctcggcgttc ggtgagaaac cactcggggg
60atgaggatct gctgctagag tgagaggagc tacggtcagt atcctctgcc ttcgtcggcg
120gcggaagtgg aggggaggaa gcgatggagg cgagcgccgg gctggtggcc ggctcccaca
180accgcaacga gctcgtcgtc atccgccgcg acggcgatcc cgggccgaag ccgccgcggg
240agcagaacgg gcaggtgtgc cagatttgcg gcgacgacgt cggccttgcc cccggcgggg
300accccttcgt ggcgtgcaac gagtgcgcct tccccgtctg ccgggactgc tacgaatacg
360agcgccggga gggcacgcag aactgccccc agtgcaagac tcgatacaag cgcctcaagg
420gctgccaacg tgtgaccggt gacgaggagg aggacggcgt cgatgacctg gacaacgagt
480tcaactggga cggccatgac tcgcagtctg tggccgagtc catgctctac ggccacatga
540gctacggccg tggaggtgac cctaatggcg cgccacaagc tttccagctc aaccccaatg
600ttccactcct caccaacggg caaatggtgg atgacatccc accggagcag cacgcgctgg
660tgccttcttt catgggtggt gggggaaaga ggatacatcc ccttccttat gcggatccca
720gcttacctgt gcaacccagg tctatggacc catccaagga tcttgctgca tatgggtatg
780gtagtgttgc ttggaaggaa cggatggaga attggaagca gagacaagag aggatgcacc
840agacggggaa tgatggtggt ggtgatgatg gtgacgatgc tgatctacca ctaatggatg
900aagcaagaca acaactgtcc aggaaaattc cacttccatc aagccagatt aatccatata
960ggatgattat cattattcgg cttgtggttt tggggttctt cttccactac cgagtgatgc
1020atccggtgaa tgatgcattt gctttgtggc tcatatctgt tatctgtgaa atctggtttg
1080ccatgtcttg gattcttgat caattcccaa agtggttccc tattgagaga gagacttacc
1140tagaccggct gtcactgagg ttcgacaagg aaggccagcc atctcaactt gctccaattg
1200atttctttgt cagtacggtt gatcccttaa aggaacctcc tttggtcaca acaaatactg
1260ttctatctat cctttcggtg gattatcctg ttgataaggt ttcttgctat gtttctgatg
1320atggtgctgc aatgctaacg tttgaagcat tatctgaaac atctgaattt gcaaagaaat
1380gggttccttt ctgcaaacgg tacaatattg aacctcgcgc tccagagtgg tacttccaac
1440agaagataga ctacttgaaa gacaaggtgg cagcaaactt tgttagggag aggagagcaa
1500tgaagagaga gtatgaggaa ttcaaggtga gaatcaatgc cttagttgcc aaagcccaga
1560aagttcctga agaaggatgg acaatgcaag atggaacccc ctggcctgga aacaatgttc
1620gtgatcatcc tggaatgatt caggtcttcc ttggccaaag cggaggcctt gactgtgagg
1680gaaatgaact gccacgattg gtttatgttt ctagagagaa acgaccaggc tataaccatc
1740ataagaaagc tggtgctatg aatgcattgg tccgagtctc tgctgtacta acaaatgctc
1800catatttgtt aaacttggat tgtgatcact acatcaacaa cagcaaggct ataaaggaag
1860caatgtgttt tatgatggac cctttactag gaaagaaggt ttgctatgta cagttccctc
1920aaagatttga tgggattgat cgccatgacc gatatgctaa ccggaatgtt gtcttttttg
1980atatcaacat gaaaggtttg gatggtattc agggtccaat ttatgttggt actggatgtg
2040tatttagaag gcaggcatta tatggttatg atgcccccaa aacaaagaag ccaccatcaa
2100ggacttgcaa ctgctggccc aagtggtgct tttgctgttg ctgctttggc aataggaagc
2160aaaagaagac taccaaaccc aaaacagaga agaaaaagtt attatttttc aagaaagaag
2220agaaccaatc ccctgcatat gctcttggtg aaattgacga agctgctcca ggagctgaga
2280atgaaaaggc cggtattgta aatcaacaaa aattagaaaa gaaatttggc caatcttctg
2340tttttgttac atccacactt ctcgagaatg gtggaacctt gaagagtgca agtcctgctt
2400ctcttttgaa agaagctata catgtcatta gttgtggtta tgaagacaag acagactggg
2460gaaaagagat tggctggatc tatggatcag ttacagaaga tattctaact ggtttcaaga
2520tgcattgtca tggttggcgg tcaatttact gcatacctaa acgggttgca ttcaaaggtt
2580ctgcacctct gaatctttca gatcgtcttc accaggtgct tcggtgggct cttgggtcta
2640ttgagatctt cttcagcaat cattgccctc tttggtatgg gtatggtggc ggtctgaaat
2700ttttggaaag attttcctac atcaactcca tcgtgtatcc ttggacatct attcccctct
2760tggcttactg tacattgcct gccatctgtt tattgacagg gaaatttatc actccagagc
2820tgaataatgt tgccagcctg tggttcatgt cactttttat ctgcattttt gctacgagca
2880tcctagaaat gagatggagt ggtgttggaa ttgatgactg gtggaggaat gagcagttct
2940gggtcattgg aggtgtgtcc tcacacctct ttgctgtgtt ccagggactt ctcaaggtca
3000tagctggtgt tgatacaagc ttcaccgtga catcaaaggg tggagatgat gaggagttct
3060cagagctata tacattcaaa tggactacct tattgatacc tcctaccacc ttgcttctat
3120tgaacttcat tggtgtggtc gctggcgttt caaatgcgat caataacgga tatgagtcat
3180ggggccccct ctttgggaag ctattctttg cattttgggt gattgtccat ctttatccct
3240ttctcaaagg tttggttgga aggcaaaaca ggacaccaac gattgtcatc gtctggtcca
3300ttctgctggc ttcaatcttc tcgctccttt gggttcggat tgatcctttc cttgcgaagg
3360atgatggtcc gcttcttgag gagtgtggtt tggattgcaa ctaggatgtc agtgcatcag
3420ctcccccaat ctgcatatgc ttgaagtata ttttctggtg tttgtcccca tattcagtgt
3480ctgtagataa gagacatgaa atgtcccaag tttcttttga tccatggtga acctacttaa
3540tatctgagag atatactggg ggaaaatgga ggctgcggca atccttgtgc agttgggccg
3600tggaatacag catatgcaag tgtttgattg tgcagcattc tttattactt ggtcgcaata
3660tagatgggct gagccgaaca gcaaggtatt ttgattctgc actgctcccg tgtacaaact
3720tggttctcaa taaggcaggc aggaatgcat ctgccagtgg aacagagcaa cctgcacatt
3780atttatgtat gcctgttcat tggagggctt gttcattaca tgttcgtcta tactagaaaa
3840aacagaatat tagcattaat ctatagttaa ttaaagtatg taaatgcgcc tgttttttgt
3900tgtgtactgt aatcatctga gttggttttg tgaaaaaaaa aaaaaaaaaa aaaaaaaaaa
3960aaaaaaaaa
3969321086PRTZea mays 32Met Glu Ala Ser Ala Gly Leu Val Ala Gly Ser His
Asn Arg Asn Glu1 5 10
15Leu Val Val Ile Arg Arg Asp Gly Asp Pro Gly Pro Lys Pro Pro Arg
20 25 30Glu Gln Asn Gly Gln Val Cys
Gln Ile Cys Gly Asp Asp Val Gly Leu 35 40
45Ala Pro Gly Gly Asp Pro Phe Val Ala Cys Asn Glu Cys Ala Phe
Pro 50 55 60Val Cys Arg Asp Cys Tyr
Glu Tyr Glu Arg Arg Glu Gly Thr Gln Asn65 70
75 80Cys Pro Gln Cys Lys Thr Arg Tyr Lys Arg Leu
Lys Gly Cys Gln Arg 85 90
95Val Thr Gly Asp Glu Glu Glu Asp Gly Val Asp Asp Leu Asp Asn Glu
100 105 110Phe Asn Trp Asp Gly His
Asp Ser Gln Ser Val Ala Glu Ser Met Leu 115 120
125Tyr Gly His Met Ser Tyr Gly Arg Gly Gly Asp Pro Asn Gly
Ala Pro 130 135 140Gln Ala Phe Gln Leu
Asn Pro Asn Val Pro Leu Leu Thr Asn Gly Gln145 150
155 160Met Val Asp Asp Ile Pro Pro Glu Gln His
Ala Leu Val Pro Ser Phe 165 170
175Met Gly Gly Gly Gly Lys Arg Ile His Pro Leu Pro Tyr Ala Asp Pro
180 185 190Ser Leu Pro Val Gln
Pro Arg Ser Met Asp Pro Ser Lys Asp Leu Ala 195
200 205Ala Tyr Gly Tyr Gly Ser Val Ala Trp Lys Glu Arg
Met Glu Asn Trp 210 215 220Lys Gln Arg
Gln Glu Arg Met His Gln Thr Gly Asn Asp Gly Gly Gly225
230 235 240Asp Asp Gly Asp Asp Ala Asp
Leu Pro Leu Met Asp Glu Ala Arg Gln 245
250 255Gln Leu Ser Arg Lys Ile Pro Leu Pro Ser Ser Gln
Ile Asn Pro Tyr 260 265 270Arg
Met Ile Ile Ile Ile Arg Leu Val Val Leu Gly Phe Phe Phe His 275
280 285Tyr Arg Val Met His Pro Val Asn Asp
Ala Phe Ala Leu Trp Leu Ile 290 295
300Ser Val Ile Cys Glu Ile Trp Phe Ala Met Ser Trp Ile Leu Asp Gln305
310 315 320Phe Pro Lys Trp
Phe Pro Ile Glu Arg Glu Thr Tyr Leu Asp Arg Leu 325
330 335Ser Leu Arg Phe Asp Lys Glu Gly Gln Pro
Ser Gln Leu Ala Pro Ile 340 345
350Asp Phe Phe Val Ser Thr Val Asp Pro Leu Lys Glu Pro Pro Leu Val
355 360 365Thr Thr Asn Thr Val Leu Ser
Ile Leu Ser Val Asp Tyr Pro Val Asp 370 375
380Lys Val Ser Cys Tyr Val Ser Asp Asp Gly Ala Ala Met Leu Thr
Phe385 390 395 400Glu Ala
Leu Ser Glu Thr Ser Glu Phe Ala Lys Lys Trp Val Pro Phe
405 410 415Cys Lys Arg Tyr Asn Ile Glu
Pro Arg Ala Pro Glu Trp Tyr Phe Gln 420 425
430Gln Lys Ile Asp Tyr Leu Lys Asp Lys Val Ala Ala Asn Phe
Val Arg 435 440 445Glu Arg Arg Ala
Met Lys Arg Glu Tyr Glu Glu Phe Lys Val Arg Ile 450
455 460Asn Ala Leu Val Ala Lys Ala Gln Lys Val Pro Glu
Glu Gly Trp Thr465 470 475
480Met Gln Asp Gly Thr Pro Trp Pro Gly Asn Asn Val Arg Asp His Pro
485 490 495Gly Met Ile Gln Val
Phe Leu Gly Gln Ser Gly Gly Leu Asp Cys Glu 500
505 510Gly Asn Glu Leu Pro Arg Leu Val Tyr Val Ser Arg
Glu Lys Arg Pro 515 520 525Gly Tyr
Asn His His Lys Lys Ala Gly Ala Met Asn Ala Leu Val Arg 530
535 540Val Ser Ala Val Leu Thr Asn Ala Pro Tyr Leu
Leu Asn Leu Asp Cys545 550 555
560Asp His Tyr Ile Asn Asn Ser Lys Ala Ile Lys Glu Ala Met Cys Phe
565 570 575Met Met Asp Pro
Leu Leu Gly Lys Lys Val Cys Tyr Val Gln Phe Pro 580
585 590Gln Arg Phe Asp Gly Ile Asp Arg His Asp Arg
Tyr Ala Asn Arg Asn 595 600 605Val
Val Phe Phe Asp Ile Asn Met Lys Gly Leu Asp Gly Ile Gln Gly 610
615 620Pro Ile Tyr Val Gly Thr Gly Cys Val Phe
Arg Arg Gln Ala Leu Tyr625 630 635
640Gly Tyr Asp Ala Pro Lys Thr Lys Lys Pro Pro Ser Arg Thr Cys
Asn 645 650 655Cys Trp Pro
Lys Trp Cys Phe Cys Cys Cys Cys Phe Gly Asn Arg Lys 660
665 670Gln Lys Lys Thr Thr Lys Pro Lys Thr Glu
Lys Lys Lys Leu Leu Phe 675 680
685Phe Lys Lys Glu Glu Asn Gln Ser Pro Ala Tyr Ala Leu Gly Glu Ile 690
695 700Asp Glu Ala Ala Pro Gly Ala Glu
Asn Glu Lys Ala Gly Ile Val Asn705 710
715 720Gln Gln Lys Leu Glu Lys Lys Phe Gly Gln Ser Ser
Val Phe Val Thr 725 730
735Ser Thr Leu Leu Glu Asn Gly Gly Thr Leu Lys Ser Ala Ser Pro Ala
740 745 750Ser Leu Leu Lys Glu Ala
Ile His Val Ile Ser Cys Gly Tyr Glu Asp 755 760
765Lys Thr Asp Trp Gly Lys Glu Ile Gly Trp Ile Tyr Gly Ser
Val Thr 770 775 780Glu Asp Ile Leu Thr
Gly Phe Lys Met His Cys His Gly Trp Arg Ser785 790
795 800Ile Tyr Cys Ile Pro Lys Arg Val Ala Phe
Lys Gly Ser Ala Pro Leu 805 810
815Asn Leu Ser Asp Arg Leu His Gln Val Leu Arg Trp Ala Leu Gly Ser
820 825 830Ile Glu Ile Phe Phe
Ser Asn His Cys Pro Leu Trp Tyr Gly Tyr Gly 835
840 845Gly Gly Leu Lys Phe Leu Glu Arg Phe Ser Tyr Ile
Asn Ser Ile Val 850 855 860Tyr Pro Trp
Thr Ser Ile Pro Leu Leu Ala Tyr Cys Thr Leu Pro Ala865
870 875 880Ile Cys Leu Leu Thr Gly Lys
Phe Ile Thr Pro Glu Leu Asn Asn Val 885
890 895Ala Ser Leu Trp Phe Met Ser Leu Phe Ile Cys Ile
Phe Ala Thr Ser 900 905 910Ile
Leu Glu Met Arg Trp Ser Gly Val Gly Ile Asp Asp Trp Trp Arg 915
920 925Asn Glu Gln Phe Trp Val Ile Gly Gly
Val Ser Ser His Leu Phe Ala 930 935
940Val Phe Gln Gly Leu Leu Lys Val Ile Ala Gly Val Asp Thr Ser Phe945
950 955 960Thr Val Thr Ser
Lys Gly Gly Asp Asp Glu Glu Phe Ser Glu Leu Tyr 965
970 975Thr Phe Lys Trp Thr Thr Leu Leu Ile Pro
Pro Thr Thr Leu Leu Leu 980 985
990Leu Asn Phe Ile Gly Val Val Ala Gly Val Ser Asn Ala Ile Asn Asn
995 1000 1005Gly Tyr Glu Ser Trp Gly
Pro Leu Phe Gly Lys Leu Phe Phe Ala 1010 1015
1020Phe Trp Val Ile Val His Leu Tyr Pro Phe Leu Lys Gly Leu
Val 1025 1030 1035Gly Arg Gln Asn Arg
Thr Pro Thr Ile Val Ile Val Trp Ser Ile 1040 1045
1050Leu Leu Ala Ser Ile Phe Ser Leu Leu Trp Val Arg Ile
Asp Pro 1055 1060 1065Phe Leu Ala Lys
Asp Asp Gly Pro Leu Leu Glu Glu Cys Gly Leu 1070
1075 1080Asp Cys Asn 1085333813DNAZea mays
33ccacagctca tataccaaga gccggagcag cttagcgcag cccagagcgg cgccgcgcca
60agcacaaccc ccacccgcca cagccgcgtg cgcatgtgag cggtcgccgc ggccgggaga
120ccagaggagg ggaggactac gtgcatttcg ctgtgccgcc gccgcggggt tcgtgcgcga
180gcgagatccg gcggggcggg gcggggggcc tgagatggag gctagcgcgg ggctggtggc
240cggctcgcat aaccggaacg agctggtggt gatccgccgc gaccgcgagt cgggagccgc
300gggcggcggc gcggcgcgcc gggcggaggc gccgtgccag atatgcggcg acgaggtcgg
360ggtgggcttc gacggggagc ccttcgtggc gtgcaacgag tgcgccttcc ccgtctgccg
420cgcctgctac gagtacgagc gccgcgaggg ctcgcaagcg tgcccgcagt gcaggacccg
480ctacaagcgc ctcaagggct gcccgcgggt ggccggcgac gaggaggagg acggcgtcga
540cgacctggag ggcgagttcg gcctgcagga cggcgccgcc cacgaggacg acccgcagta
600cgtcgccgag tccatgctca gggcgcagat gagctacggc cgcggcggcg acgcgcaccc
660cggcttcagc cccgtcccca acgtgccgct cctcaccaac ggccagatgg ttgatgacat
720cccgccggag cagcacgcgc tcgtgccgtc ctacatgagc ggcggcggcg gcgggggcaa
780gaggatccac ccgctccctt tcgcagatcc caaccttcca gtgcaaccga gatccatgga
840cccgtccaag gatctggccg cctacggata tggcagcgtg gcctggaagg agagaatgga
900gggctggaag cagaagcagg agcgcctgca gcatgtcagg agcgagggtg gcggtgattg
960ggatggcgac gatgcagatc tgccactaat ggatgaagct aggcagccat tgtccagaaa
1020agtccctata tcatcaagcc gaattaatcc ctacaggatg attatcgtta tccggttggt
1080ggttttgggt ttcttcttcc actaccgagt gatgcatccg gcgaaagatg catttgcatt
1140gtggctcata tctgtaatct gtgaaatctg gtttgcgatg tcctggattc ttgatcagtt
1200cccaaagtgg cttccaatcg agagagagac ttacctggac cgtttgtcac taaggtttga
1260caaggaaggt caaccctctc agcttgctcc aatcgacttc tttgtcagta cggttgatcc
1320cacaaaggaa cctcccttgg tcacagcgaa cactgtcctt tccatccttt ctgtggatta
1380tccggttgag aaggtctcct gctatgtttc tgatgatggt gctgcaatgc ttacgtttga
1440agcattgtct gaaacatctg aatttgcaaa gaaatgggtt cctttcagca aaaagtttaa
1500tatcgagcct cgtgctcctg agtggtactt ccaacagaag atagactacc tgaaagacaa
1560ggttgctgct tcatttgtta gggagaggag ggcgatgaag agagaatacg aggaattcaa
1620ggtaaggatc aatgccttgg ttgcaaaagc ccaaaaggtt cctgaggaag gatggacaat
1680gcaagatgga agcccctggc ctggaaacaa cgtacgcgat catcctggaa tgattcaggt
1740attccttggc caaagtggcg gtcgtgatgt ggaaggaaat gagttgcctc gcctggttta
1800tgtctcgaga gaaaagaggc caggttataa ccatcacaag aaggctggtg ccatgaatgc
1860actggtccgt gtctctgctg tcttatcaaa tgctgcatac ctattgaact tggactgtga
1920tcactacatc aacaatagca aggccataaa agaggctatg tgtttcatga tggatccttt
1980ggtggggaag aaagtgtgct atgtacagtt ccctcagagg tttgatggta ttgacaaaaa
2040tgatcgatac gctaacagga acgttgtctt ttttgacatc aacatgaaag gtttggacgg
2100tattcaagga cccatttatg tgggtactgg atgtgttttc agacggcagg cactgtatgg
2160ttatgatgct cctaaaacga agaagccacc atcaagaact tgcaactgct ggcccaagtg
2220gtgcctctct tgctgctgca gcaggaacaa gaataaaaag aagactacaa aaccaaagac
2280ggagaagaag aaaagattat ttttcaagaa agcagaaaac ccatctcctg catatgcttt
2340gggtgaaatt gatgaaggtg ctccaggtgc tgatatcgag aaggccggaa tcgtaaatca
2400acagaaacta gagaagaaat ttgggcagtc ttctgttttt gtcgcatcaa cacttcttga
2460gaacggaggg accctgaaga gcgcaagtcc agcttctctt ctgaaggaag ctatacatgt
2520tatcagctgc ggctacgaag acaagaccga ctggggaaaa gagattggct ggatttacgg
2580atcgatcaca gaggatatct tgactggatt taagatgcac tgccatggct ggcggtctat
2640ttactgcatc ccgaagcggc ctgcattcaa aggttctgcg cctctgaacc tttccgaccg
2700tcttcaccag gtccttcgct gggcccttgg gtccgtcgaa attttcttca gcaagcactg
2760cccactttgg tacggatacg gcggcgggct aaaattcctg gaaaggtttt cttatatcaa
2820ctccatcgtt tatccctgga cgtccattcc tctcctggct tactgtacct tgcctgccat
2880ctgcctgctc acggggaagt ttatcacacc agagcttacc aatgtcgcca gtatctggtt
2940catggcactt ttcatctgca tctccgtgac cggcatcctg gaaatgaggt ggagtggcgt
3000ggccatcgac gactggtgga ggaacgagca gttctgggtc atcggaggcg tttcggcgca
3060tctgttcgcg gtgttccagg gcctgctgaa ggtgttcgcc ggcatcgaca cgagcttcac
3120cgtgacgtcg aaggccgggg acgacgagga gttctcggag ctgtacacgt tcaagtggac
3180caccctgctg atacccccga ccacgctcct cctgctgaac ttcatcgggg tggtggccgg
3240gatctcgaac gcgatcaaca acgggtacga gtcgtggggc cccctgttcg ggaagctctt
3300cttcgccttc tgggtgatcg tccacctgta cccgttcctc aagggtctgg tggggaggca
3360gaacaggacg ccgacgatcg tcatcgtctg gtccatcctg ctggcctcga tcttctcgct
3420cctgtgggtc cgcgtcgacc cgttcctcgc caagagcaac ggcccgctcc tggaggagtg
3480tggcctggac tgcaactgaa gtgggggccc cctgtcactc gaagttctgt cacgggcgaa
3540ttacgcctga ttttttgttg ttgttgttgt tggaattctt tgctgtagat agaaaccaca
3600tgtccacggc atctctgctg tgtccattgg agcaggagag aggtgcctgc tgctgtttgt
3660tgagtaaatt aaaagtttta aagttataca gtgatgcaca ttccagtgcc cagtgtattc
3720cctttttaca gtctgtatat tagcgacaaa ggacatattg gttaggagtt tgattctttt
3780gtaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaa
3813341094PRTZea mays 34Met Glu Ala Ser Ala Gly Leu Val Ala Gly Ser His
Asn Arg Asn Glu1 5 10
15Leu Val Val Ile Arg Arg Asp Arg Glu Ser Gly Ala Ala Gly Gly Gly
20 25 30Ala Ala Arg Arg Ala Glu Ala
Pro Cys Gln Ile Cys Gly Asp Glu Val 35 40
45Gly Val Gly Phe Asp Gly Glu Pro Phe Val Ala Cys Asn Glu Cys
Ala 50 55 60Phe Pro Val Cys Arg Ala
Cys Tyr Glu Tyr Glu Arg Arg Glu Gly Ser65 70
75 80Gln Ala Cys Pro Gln Cys Arg Thr Arg Tyr Lys
Arg Leu Lys Gly Cys 85 90
95Pro Arg Val Ala Gly Asp Glu Glu Glu Asp Gly Val Asp Asp Leu Glu
100 105 110Gly Glu Phe Gly Leu Gln
Asp Gly Ala Ala His Glu Asp Asp Pro Gln 115 120
125Tyr Val Ala Glu Ser Met Leu Arg Ala Gln Met Ser Tyr Gly
Arg Gly 130 135 140Gly Asp Ala His Pro
Gly Phe Ser Pro Val Pro Asn Val Pro Leu Leu145 150
155 160Thr Asn Gly Gln Met Val Asp Asp Ile Pro
Pro Glu Gln His Ala Leu 165 170
175Val Pro Ser Tyr Met Ser Gly Gly Gly Gly Gly Gly Lys Arg Ile His
180 185 190Pro Leu Pro Phe Ala
Asp Pro Asn Leu Pro Val Gln Pro Arg Ser Met 195
200 205Asp Pro Ser Lys Asp Leu Ala Ala Tyr Gly Tyr Gly
Ser Val Ala Trp 210 215 220Lys Glu Arg
Met Glu Gly Trp Lys Gln Lys Gln Glu Arg Leu Gln His225
230 235 240Val Arg Ser Glu Gly Gly Gly
Asp Trp Asp Gly Asp Asp Ala Asp Leu 245
250 255Pro Leu Met Asp Glu Ala Arg Gln Pro Leu Ser Arg
Lys Val Pro Ile 260 265 270Ser
Ser Ser Arg Ile Asn Pro Tyr Arg Met Ile Ile Val Ile Arg Leu 275
280 285Val Val Leu Gly Phe Phe Phe His Tyr
Arg Val Met His Pro Ala Lys 290 295
300Asp Ala Phe Ala Leu Trp Leu Ile Ser Val Ile Cys Glu Ile Trp Phe305
310 315 320Ala Met Ser Trp
Ile Leu Asp Gln Phe Pro Lys Trp Leu Pro Ile Glu 325
330 335Arg Glu Thr Tyr Leu Asp Arg Leu Ser Leu
Arg Phe Asp Lys Glu Gly 340 345
350Gln Pro Ser Gln Leu Ala Pro Ile Asp Phe Phe Val Ser Thr Val Asp
355 360 365Pro Thr Lys Glu Pro Pro Leu
Val Thr Ala Asn Thr Val Leu Ser Ile 370 375
380Leu Ser Val Asp Tyr Pro Val Glu Lys Val Ser Cys Tyr Val Ser
Asp385 390 395 400Asp Gly
Ala Ala Met Leu Thr Phe Glu Ala Leu Ser Glu Thr Ser Glu
405 410 415Phe Ala Lys Lys Trp Val Pro
Phe Ser Lys Lys Phe Asn Ile Glu Pro 420 425
430Arg Ala Pro Glu Trp Tyr Phe Gln Gln Lys Ile Asp Tyr Leu
Lys Asp 435 440 445Lys Val Ala Ala
Ser Phe Val Arg Glu Arg Arg Ala Met Lys Arg Glu 450
455 460Tyr Glu Glu Phe Lys Val Arg Ile Asn Ala Leu Val
Ala Lys Ala Gln465 470 475
480Lys Val Pro Glu Glu Gly Trp Thr Met Gln Asp Gly Ser Pro Trp Pro
485 490 495Gly Asn Asn Val Arg
Asp His Pro Gly Met Ile Gln Val Phe Leu Gly 500
505 510Gln Ser Gly Gly Arg Asp Val Glu Gly Asn Glu Leu
Pro Arg Leu Val 515 520 525Tyr Val
Ser Arg Glu Lys Arg Pro Gly Tyr Asn His His Lys Lys Ala 530
535 540Gly Ala Met Asn Ala Leu Val Arg Val Ser Ala
Val Leu Ser Asn Ala545 550 555
560Ala Tyr Leu Leu Asn Leu Asp Cys Asp His Tyr Ile Asn Asn Ser Lys
565 570 575Ala Ile Lys Glu
Ala Met Cys Phe Met Met Asp Pro Leu Val Gly Lys 580
585 590Lys Val Cys Tyr Val Gln Phe Pro Gln Arg Phe
Asp Gly Ile Asp Lys 595 600 605Asn
Asp Arg Tyr Ala Asn Arg Asn Val Val Phe Phe Asp Ile Asn Met 610
615 620Lys Gly Leu Asp Gly Ile Gln Gly Pro Ile
Tyr Val Gly Thr Gly Cys625 630 635
640Val Phe Arg Arg Gln Ala Leu Tyr Gly Tyr Asp Ala Pro Lys Thr
Lys 645 650 655Lys Pro Pro
Ser Arg Thr Cys Asn Cys Trp Pro Lys Trp Cys Leu Ser 660
665 670Cys Cys Cys Ser Arg Asn Lys Asn Lys Lys
Lys Thr Thr Lys Pro Lys 675 680
685Thr Glu Lys Lys Lys Arg Leu Phe Phe Lys Lys Ala Glu Asn Pro Ser 690
695 700Pro Ala Tyr Ala Leu Gly Glu Ile
Asp Glu Gly Ala Pro Gly Ala Asp705 710
715 720Ile Glu Lys Ala Gly Ile Val Asn Gln Gln Lys Leu
Glu Lys Lys Phe 725 730
735Gly Gln Ser Ser Val Phe Val Ala Ser Thr Leu Leu Glu Asn Gly Gly
740 745 750Thr Leu Lys Ser Ala Ser
Pro Ala Ser Leu Leu Lys Glu Ala Ile His 755 760
765Val Ile Ser Cys Gly Tyr Glu Asp Lys Thr Asp Trp Gly Lys
Glu Ile 770 775 780Gly Trp Ile Tyr Gly
Ser Ile Thr Glu Asp Ile Leu Thr Gly Phe Lys785 790
795 800Met His Cys His Gly Trp Arg Ser Ile Tyr
Cys Ile Pro Lys Arg Pro 805 810
815Ala Phe Lys Gly Ser Ala Pro Leu Asn Leu Ser Asp Arg Leu His Gln
820 825 830Val Leu Arg Trp Ala
Leu Gly Ser Val Glu Ile Phe Phe Ser Lys His 835
840 845Cys Pro Leu Trp Tyr Gly Tyr Gly Gly Gly Leu Lys
Phe Leu Glu Arg 850 855 860Phe Ser Tyr
Ile Asn Ser Ile Val Tyr Pro Trp Thr Ser Ile Pro Leu865
870 875 880Leu Ala Tyr Cys Thr Leu Pro
Ala Ile Cys Leu Leu Thr Gly Lys Phe 885
890 895Ile Thr Pro Glu Leu Thr Asn Val Ala Ser Ile Trp
Phe Met Ala Leu 900 905 910Phe
Ile Cys Ile Ser Val Thr Gly Ile Leu Glu Met Arg Trp Ser Gly 915
920 925Val Ala Ile Asp Asp Trp Trp Arg Asn
Glu Gln Phe Trp Val Ile Gly 930 935
940Gly Val Ser Ala His Leu Phe Ala Val Phe Gln Gly Leu Leu Lys Val945
950 955 960Phe Ala Gly Ile
Asp Thr Ser Phe Thr Val Thr Ser Lys Ala Gly Asp 965
970 975Asp Glu Glu Phe Ser Glu Leu Tyr Thr Phe
Lys Trp Thr Thr Leu Leu 980 985
990Ile Pro Pro Thr Thr Leu Leu Leu Leu Asn Phe Ile Gly Val Val Ala
995 1000 1005Gly Ile Ser Asn Ala Ile
Asn Asn Gly Tyr Glu Ser Trp Gly Pro 1010 1015
1020Leu Phe Gly Lys Leu Phe Phe Ala Phe Trp Val Ile Val His
Leu 1025 1030 1035Tyr Pro Phe Leu Lys
Gly Leu Val Gly Arg Gln Asn Arg Thr Pro 1040 1045
1050Thr Ile Val Ile Val Trp Ser Ile Leu Leu Ala Ser Ile
Phe Ser 1055 1060 1065Leu Leu Trp Val
Arg Val Asp Pro Phe Leu Ala Lys Ser Asn Gly 1070
1075 1080Pro Leu Leu Glu Glu Cys Gly Leu Asp Cys Asn
1085 1090353799DNAZea maysmisc_feature(3757)..(3757)n is
a, c, g, or t 35caactcacgt tgccgcggct tcctccatcg gtgcggtgcc ctgtcctttt
ctctcctcca 60cctccctagt ccctcctccc ccccgcatac atagctacta ctagtagcac
cacgctcgca 120gcgggagatg cggtgctgat ccgtgcccct gctcggatct cgggagtggt
gccgacttgt 180gtcgcttcgg ctctgcctag gccagctcct tgtcggttct gggcgagctc
gcctgccatg 240gagggcgacg cggacggcgt gaagtcgggg aggcgcgggg gagggcaggt
gtgccagatc 300tgcggcgatg gcgtgggcac tacggcggag ggagacgtct tcaccgcctg
cgacgtctgc 360gggttcccgg tgtgccgccc ctgctacgag tacgagcgca aggacggcac
acaagcgtgc 420ccccagtgca aaaacaagta caagcgccac aaggggagtc cagcgatccg
aggggaggaa 480ggagacgata ctgatgccga tgatgctagc gacttcaact accctgcatc
tggcaatgac 540gaccagaagc agaagattgc tgacaggatg cgcagctggc gcatgaatgc
tgggggcagc 600ggggatgttg gccgccccaa gtatgacagt ggtgagatcg ggcttaccaa
gtacgacagt 660ggtgagatcc ctcggggata catcccgtca gtcactaaca gccagatttc
gggagaaatc 720cctggtgctt cccctgacca tcatatgatg tctcctactg ggaacattgg
caggcgcgcc 780ccatttccct atatgaatca ttcatcaaat ccgtcgaggg aattctctgg
tagcgttggg 840aatgttgcct ggaaagagag ggttgatggc tggaaaatga agcaggacaa
gggaacaatt 900cccatgacga atggcacaag cattgctccc tctgagggcc ggggtgttgg
tgatattgat 960gcatcaactg attacaacat ggaagatgcc ttattaaacg atgaaactcg
ccagcctcta 1020tctaggaaag ttccacttcc ttcctccagg ataaatccat acaggatggt
cattgtgcta 1080cgattgattg ttctaagcat cttcttgcac taccggatca caaatcctgt
gcgtaatgca 1140tacccactgt ggcttctatc tgttatatgt gagatctggt ttgctctttc
ctggatattg 1200gatcagtttc caaagtggtt tccaatcaac cgcgagactt accttgatag
actcgcatta 1260aggtatgacc gggaaggtga gccatctcag ttggctgctg ttgacatttt
tgtcagtact 1320gtcgacccaa tgaaggagcc tcctcttgtc actgccaata ccgtgctatc
cattctcgct 1380gtggactatc ctgtggataa ggtctcttgc tatgtatctg atgatggagc
tgctatgctg 1440acatttgatg cactagctga gacttcagag tttgctagaa aatgggtgcc
atttgttaag 1500aagtacaaca ttgaacctag agctcctgaa tggtacttct cccagaaaat
tgattacttg 1560aaggacaaag tgcacccttc atttgttaaa gaccgccggg ccatgaagag
agaatatgaa 1620gaattcaaaa ttagggtaaa tggccttgtt gctaaggcac aaaaagtccc
tgaggaagga 1680tggatcatgc aagatggcac accatggcca ggaaacaata ccagggacca
tcctggaatg 1740attcaggttt tccttggtca cagtggtggt cttgatactg agggtaatga
gctaccccgt 1800ttggtctatg tttctcgtga aaaacgtcct ggattccagc atcacaagaa
agctggtgcc 1860atgaatgctc ttgtccgcgt ctcagctgtg cttaccaatg gacaatacat
gttgaatctt 1920gattgtgatc actacatcaa caacagtaag gctctcaggg aagctatgtg
cttccttatg 1980gatcctaacc taggaaggag tgtctgctat gttcagtttc cccagaggtt
cgatggtatt 2040gataggaatg atcgatatgc caacaggaac accgtgtttt tcgatattaa
cttgagaggt 2100cttgatggca tccaaggacc agtttatgtg ggcactggct gtgttttcaa
cagaacagct 2160ctatatggtt atgagccccc aattaagcaa aagaagggtg gtttcttgtc
atcactatgt 2220ggtggcagga agaagggaag caaatcaaag aagggctcag acaagaaaaa
gtcacagaag 2280catgtggaca gttctgtgcc agtattcaat cttgaagata tagaggaggg
agttgaaggc 2340gctggatttg atgatgagaa atcacttctt atgtctcaaa tgagcttgga
gaagagattt 2400ggccaatctg cagcttttgt tgcgtccact ctgatggaat atggtggtgt
tcctcagtct 2460gcgactccag aatctcttct gaaagaagct atccatgtca taagttgtgg
ctacgaggac 2520aagattgaat ggggaactga gattgggtgg atctatggtt ctgtgacgga
agatattctc 2580actgggttca agatgcacgc acgaggctgg cggtcgatct actgcatgcc
taagcggccg 2640gccttcaagg gatcggctcc catcaatctc tcagaccgtc tgaaccaggt
gctccggtgg 2700gctctcggtt cagtggaaat ccttttcagc cggcattgcc ccctatggta
cgggtacgga 2760ggacgcctga agttcttgga gagattcgcc tacatcaaca ccaccatcta
cccgctcacg 2820tccctcccgc tcctcattta ctgtatcctg cctgccatct gcctgctcac
ggggaagttc 2880atcatcccag agatcagcaa cttcgctagt atctggttca tctctctctt
catctcgatc 2940ttcgccacgg gtatcctgga gatgaggtgg agcggcgtgg gcatcgacga
gtggtggagg 3000aacgagcagt tctgggtcat cggaggcatc tccgcccacc tcttcgccgt
cttccagggc 3060ctcctcaagg tgcttgccgg catcgacacc aacttcaccg tcacctccaa
ggcctcggat 3120gaagacggcg acttcgcgga gctgtacatg ttcaagtgga cgacacttct
gatcccgccc 3180accaccatcc tgatcatcaa cctggtcggc gttgttgccg gcatctccta
cgccatcaac 3240agcgggtacc agtcgtgggg tccgctcttc ggcaagctct tcttcgcctt
ctgggtgatc 3300gttcacctgt acccgttcct caagggtctc atgggtcggc agaaccgcac
cccgaccatc 3360gtggttgtct gggcgatcct gctggcgtcg atcttctcct tgctgtgggt
tcgcatcgat 3420ccgttcacca accgcgtcac tggcccggat actcgaacgt gtggcatcaa
ctgctaggga 3480ggtggaaggt ttgtagaaac agagagatac cacgaatgtg ccgctgccac
aaattgtctg 3540ttagtaagtt atataggcag gtggcgttat ttacagctac gtacacacaa
ggggatactc 3600cgtttatcac tggtgtgcat tcttttgttg atataagtta ctatatatac
gtattgcttc 3660tactttgtgg agagtggctg acaggaccag ttttgtaatg ttatgaacag
caaagaaata 3720agttagtttc caaaaaaaaa aaaaaaaaaa aaaaaanaaa aaaaaaaaaa
aaaananaaa 3780anaaaaaaaa aaaaacccc
3799361079PRTZea mays 36Met Glu Gly Asp Ala Asp Gly Val Lys
Ser Gly Arg Arg Gly Gly Gly1 5 10
15Gln Val Cys Gln Ile Cys Gly Asp Gly Val Gly Thr Thr Ala Glu
Gly 20 25 30Asp Val Phe Thr
Ala Cys Asp Val Cys Gly Phe Pro Val Cys Arg Pro 35
40 45Cys Tyr Glu Tyr Glu Arg Lys Asp Gly Thr Gln Ala
Cys Pro Gln Cys 50 55 60Lys Asn Lys
Tyr Lys Arg His Lys Gly Ser Pro Ala Ile Arg Gly Glu65 70
75 80Glu Gly Asp Asp Thr Asp Ala Asp
Asp Ala Ser Asp Phe Asn Tyr Pro 85 90
95Ala Ser Gly Asn Asp Asp Gln Lys Gln Lys Ile Ala Asp Arg
Met Arg 100 105 110Ser Trp Arg
Met Asn Ala Gly Gly Ser Gly Asp Val Gly Arg Pro Lys 115
120 125Tyr Asp Ser Gly Glu Ile Gly Leu Thr Lys Tyr
Asp Ser Gly Glu Ile 130 135 140Pro Arg
Gly Tyr Ile Pro Ser Val Thr Asn Ser Gln Ile Ser Gly Glu145
150 155 160Ile Pro Gly Ala Ser Pro Asp
His His Met Met Ser Pro Thr Gly Asn 165
170 175Ile Gly Arg Arg Ala Pro Phe Pro Tyr Met Asn His
Ser Ser Asn Pro 180 185 190Ser
Arg Glu Phe Ser Gly Ser Val Gly Asn Val Ala Trp Lys Glu Arg 195
200 205Val Asp Gly Trp Lys Met Lys Gln Asp
Lys Gly Thr Ile Pro Met Thr 210 215
220Asn Gly Thr Ser Ile Ala Pro Ser Glu Gly Arg Gly Val Gly Asp Ile225
230 235 240Asp Ala Ser Thr
Asp Tyr Asn Met Glu Asp Ala Leu Leu Asn Asp Glu 245
250 255Thr Arg Gln Pro Leu Ser Arg Lys Val Pro
Leu Pro Ser Ser Arg Ile 260 265
270Asn Pro Tyr Arg Met Val Ile Val Leu Arg Leu Ile Val Leu Ser Ile
275 280 285Phe Leu His Tyr Arg Ile Thr
Asn Pro Val Arg Asn Ala Tyr Pro Leu 290 295
300Trp Leu Leu Ser Val Ile Cys Glu Ile Trp Phe Ala Leu Ser Trp
Ile305 310 315 320Leu Asp
Gln Phe Pro Lys Trp Phe Pro Ile Asn Arg Glu Thr Tyr Leu
325 330 335Asp Arg Leu Ala Leu Arg Tyr
Asp Arg Glu Gly Glu Pro Ser Gln Leu 340 345
350Ala Ala Val Asp Ile Phe Val Ser Thr Val Asp Pro Met Lys
Glu Pro 355 360 365Pro Leu Val Thr
Ala Asn Thr Val Leu Ser Ile Leu Ala Val Asp Tyr 370
375 380Pro Val Asp Lys Val Ser Cys Tyr Val Ser Asp Asp
Gly Ala Ala Met385 390 395
400Leu Thr Phe Asp Ala Leu Ala Glu Thr Ser Glu Phe Ala Arg Lys Trp
405 410 415Val Pro Phe Val Lys
Lys Tyr Asn Ile Glu Pro Arg Ala Pro Glu Trp 420
425 430Tyr Phe Ser Gln Lys Ile Asp Tyr Leu Lys Asp Lys
Val His Pro Ser 435 440 445Phe Val
Lys Asp Arg Arg Ala Met Lys Arg Glu Tyr Glu Glu Phe Lys 450
455 460Ile Arg Val Asn Gly Leu Val Ala Lys Ala Gln
Lys Val Pro Glu Glu465 470 475
480Gly Trp Ile Met Gln Asp Gly Thr Pro Trp Pro Gly Asn Asn Thr Arg
485 490 495Asp His Pro Gly
Met Ile Gln Val Phe Leu Gly His Ser Gly Gly Leu 500
505 510Asp Thr Glu Gly Asn Glu Leu Pro Arg Leu Val
Tyr Val Ser Arg Glu 515 520 525Lys
Arg Pro Gly Phe Gln His His Lys Lys Ala Gly Ala Met Asn Ala 530
535 540Leu Val Arg Val Ser Ala Val Leu Thr Asn
Gly Gln Tyr Met Leu Asn545 550 555
560Leu Asp Cys Asp His Tyr Ile Asn Asn Ser Lys Ala Leu Arg Glu
Ala 565 570 575Met Cys Phe
Leu Met Asp Pro Asn Leu Gly Arg Ser Val Cys Tyr Val 580
585 590Gln Phe Pro Gln Arg Phe Asp Gly Ile Asp
Arg Asn Asp Arg Tyr Ala 595 600
605Asn Arg Asn Thr Val Phe Phe Asp Ile Asn Leu Arg Gly Leu Asp Gly 610
615 620Ile Gln Gly Pro Val Tyr Val Gly
Thr Gly Cys Val Phe Asn Arg Thr625 630
635 640Ala Leu Tyr Gly Tyr Glu Pro Pro Ile Lys Gln Lys
Lys Gly Gly Phe 645 650
655Leu Ser Ser Leu Cys Gly Gly Arg Lys Lys Gly Ser Lys Ser Lys Lys
660 665 670Gly Ser Asp Lys Lys Lys
Ser Gln Lys His Val Asp Ser Ser Val Pro 675 680
685Val Phe Asn Leu Glu Asp Ile Glu Glu Gly Val Glu Gly Ala
Gly Phe 690 695 700Asp Asp Glu Lys Ser
Leu Leu Met Ser Gln Met Ser Leu Glu Lys Arg705 710
715 720Phe Gly Gln Ser Ala Ala Phe Val Ala Ser
Thr Leu Met Glu Tyr Gly 725 730
735Gly Val Pro Gln Ser Ala Thr Pro Glu Ser Leu Leu Lys Glu Ala Ile
740 745 750His Val Ile Ser Cys
Gly Tyr Glu Asp Lys Ile Glu Trp Gly Thr Glu 755
760 765Ile Gly Trp Ile Tyr Gly Ser Val Thr Glu Asp Ile
Leu Thr Gly Phe 770 775 780Lys Met His
Ala Arg Gly Trp Arg Ser Ile Tyr Cys Met Pro Lys Arg785
790 795 800Pro Ala Phe Lys Gly Ser Ala
Pro Ile Asn Leu Ser Asp Arg Leu Asn 805
810 815Gln Val Leu Arg Trp Ala Leu Gly Ser Val Glu Ile
Leu Phe Ser Arg 820 825 830His
Cys Pro Leu Trp Tyr Gly Tyr Gly Gly Arg Leu Lys Phe Leu Glu 835
840 845Arg Phe Ala Tyr Ile Asn Thr Thr Ile
Tyr Pro Leu Thr Ser Leu Pro 850 855
860Leu Leu Ile Tyr Cys Ile Leu Pro Ala Ile Cys Leu Leu Thr Gly Lys865
870 875 880Phe Ile Ile Pro
Glu Ile Ser Asn Phe Ala Ser Ile Trp Phe Ile Ser 885
890 895Leu Phe Ile Ser Ile Phe Ala Thr Gly Ile
Leu Glu Met Arg Trp Ser 900 905
910Gly Val Gly Ile Asp Glu Trp Trp Arg Asn Glu Gln Phe Trp Val Ile
915 920 925Gly Gly Ile Ser Ala His Leu
Phe Ala Val Phe Gln Gly Leu Leu Lys 930 935
940Val Leu Ala Gly Ile Asp Thr Asn Phe Thr Val Thr Ser Lys Ala
Ser945 950 955 960Asp Glu
Asp Gly Asp Phe Ala Glu Leu Tyr Met Phe Lys Trp Thr Thr
965 970 975Leu Leu Ile Pro Pro Thr Thr
Ile Leu Ile Ile Asn Leu Val Gly Val 980 985
990Val Ala Gly Ile Ser Tyr Ala Ile Asn Ser Gly Tyr Gln Ser
Trp Gly 995 1000 1005Pro Leu Phe
Gly Lys Leu Phe Phe Ala Phe Trp Val Ile Val His 1010
1015 1020Leu Tyr Pro Phe Leu Lys Gly Leu Met Gly Arg
Gln Asn Arg Thr 1025 1030 1035Pro Thr
Ile Val Val Val Trp Ala Ile Leu Leu Ala Ser Ile Phe 1040
1045 1050Ser Leu Leu Trp Val Arg Ile Asp Pro Phe
Thr Asn Arg Val Thr 1055 1060 1065Gly
Pro Asp Thr Arg Thr Cys Gly Ile Asn Cys 1070
1075373470DNAZea mays 37gcccccggtc gatcgctcgg caatcggcat ggacgccggc
tcggtcaccg gtggcctcgc 60cgcgggctcg cacatgcggg acgagctgca tgtcatgcgc
gcccgcgagg agccgaacgc 120caaggtccgg agcgccgacg tgaagacgtg ccgcgtgtgc
gccgacgagg tcgggacgcg 180ggaggacggg cagcccttcg tggcgtgcgc cgagtgcggc
ttccccgtct gccggccctg 240ctacgagtac gagcgcagcg agggcacgca gtgctgcccg
cagtgcaaca cccgctacaa 300gcgccagaaa gggtgcccga gggtggaagg ggacgaggag
gagggcccgg agatggacga 360cttcgaggac gagttccccg ccaagagccc caagaagcct
cacgagcctg tcgcgttcga 420cgtctactcg gagaacggcg agcacccggc gcagaaatgg
cggacgggtg gccagacgct 480gtcgtccttc accggaagcg tcgccgggaa ggacctggag
gcggagaggg agatggaggg 540gagcatggag tggaaggacc ggatcgacaa gtggaagacc
aagcaggaga agaggggcaa 600gctcaaccac gacgacagcg acgacgacga cgacaagaac
gaagacgagt acatgctgct 660tgccgaggcc cgacagccgc tgtggcgcaa ggttccgatc
ccgtcgagca tgatcaaccc 720gtaccgcatc gtcatcgtgc tccgcctggt ggtgctctgc
ttcttcctca agttccggat 780cacgacgccc gccacggacg ccgtgcctct gtggctggcg
tccgtcatct gcgagctctg 840gttcgccttc tcctggatcc tggaccagct gccaaagtgg
gcgccggtga cgcgggagac 900gtacctggac cgcctggcgc tgcggtacga ccgtgagggc
gaggcgtgcc ggctgtcccc 960catcgacttc ttcgtcagca cggtggaccc gctcaaggag
ccgcccatca tcaccgccaa 1020caccgtgctg tccatcctcg ccgtcgacta ccccgtggac
cgcgtcagct gctacgtctc 1080cgacgacggc gcgtccatgc tgctcttcga cgcgctgtcc
gagaccgccg agttcgcgcg 1140ccgctgggtg cccttctgca agaagttcgc cgtggagccg
cgcgccccgg agttctactt 1200ctcgcagaag atcgactacc tcaaggacaa ggtgcagccg
acgttcgtca aggagcgccg 1260cgccatgaag agggagtacg aggagttcaa ggtgcgcatc
aacgcgctgg tggccaaggc 1320gcagaagaag cccgaggagg ggtgggtcat gcaggacggc
acgccgtggc ccgggaacaa 1380cacgcgcgac cacccgggta tgatccaggt ctacctcggc
aaccagggcg cgctggacgt 1440ggagggccac gagctgccgc gcctcgtcta cgtgtcccgt
gagaagcgcc ccgggtacaa 1500ccaccacaag aaggcgggcg ccatgaacgc gctggtgcgc
gtctccgccg tgctcaccaa 1560cgcgcccttc atcctcaacc tcgactgcga ccactacgtc
aacaacagca aggccgtgcg 1620cgaggccatg tgcttcctca tggacccgca gctggggaag
aagctctgct acgtccagtt 1680cccgcagcgc ttcgatggca tcgatcgcca cgaccgatac
gccaaccgca acgtcgtctt 1740cttcgacatc aacatgaagg ggctggacgg catccagggc
ccggtgtacg tcggcacggg 1800gtgcgtgttc aaccgccagg cgctgtacgg ctacgacccg
ccgcggcccg agaagcggcc 1860caagatgacg tgcgactgct ggccgtcgtg gtgctgctgc
tgctgctgct tcggcggcgg 1920caagcgcggc aaggcgcgca aggacaagaa gggcgacggc
ggcgaggagc cgcgccgggg 1980cctgctcggc ttctacagga agcggagcaa gaaggacaag
ctcggcggcg ggtcggtggc 2040cggcagcaag aagggcggcg ggctgtacaa gaagcaccag
cgcgcgttcg agctggagga 2100gatcgaggag gggctggagg ggtacgacga gctggagcgc
tcctcgctca tgtcgcagaa 2160gagcttcgag aagcggttcg gccagtcgcc cgtgttcatc
gcctccacgc tcgtcgagga 2220cggcggcctg ccgcagggcg ccgccgccga ccccgccgcg
ctcatcaagg aggccatcca 2280cgtcatcagc tgcggatacg aggagaagac cgagtggggc
aaggagattg ggtggatcta 2340tgggtcggtg acagaggata tcctgacggg gttcaagatg
cactgccggg ggtggaagtc 2400cgtgtactgc acgccgacac ggccggcgtt caaggggtcg
gcgcccatca acttgtctga 2460tcgtctccac caggtgctgc gctgggcgct ggggtccgtg
gagatcttca tgagccgcca 2520ctgcccgctc cggtacgcct acggcggccg gctcaagtgg
ctggagcgct tcgcctacac 2580caacaccatc gtgtacccct tcacctccat cccgctcctc
gcctactgca ccatccccgc 2640cgtctgcctg ctcaccggca agttcatcat tcccacgctg
aacaacctcg ccagcatctg 2700gttcatcgcg ctcttcctgt ccatcatcgc gacgagcgtc
ctggagctgc ggtggagcgg 2760ggtgagcatc gaggactggt ggcgcaacga gcagttctgg
gtcatcggcg gcgtgtccgc 2820gcatctcttc gccgtgttcc agggcttcct caaggttctg
ggcggcgtgg acaccagctt 2880caccgtcacc tccaaggcgg ccggcgacga ggccgacgcc
ttcggggacc tctacctctt 2940caagtggacc accctgctgg tgccccccac cacgctcatc
atcatcaaca tggtgggcat 3000cgtggccggc gtgtccgacg ccgtcaacaa cggctacggc
tcctggggcc cgctcttcgg 3060caagctcttc ttctccttct gggtcatcgt ccacctctac
ccgttcctca aggggctcat 3120ggggaggcag aaccggacgc ccaccatcgt cgtgctctgg
tccatcctcc tcgcctccat 3180cttctcgctc gtctgggtca ggatcgaccc gtttatcccg
aaggccaagg gccccatcct 3240caagccatgc ggagtcgagt gctgagctca cctagctacc
ttcttgttgc atgtacggac 3300gccgccgtgc gtttggacat acaggcactt ttgggccagg
ctactcatgt tcgacttttt 3360ttttaatttt gtacaagatt tgtgatcgag tgactgagtg
agacagagtg ttgggtgtaa 3420gaactgtgat ggaattcact caaattaatg gacatttttt
ttcttcaaaa 3470381078PRTZea mays 38Met Asp Ala Gly Ser Val
Thr Gly Gly Leu Ala Ala Gly Ser His Met1 5
10 15Arg Asp Glu Leu His Val Met Arg Ala Arg Glu Glu
Pro Asn Ala Lys 20 25 30Val
Arg Ser Ala Asp Val Lys Thr Cys Arg Val Cys Ala Asp Glu Val 35
40 45Gly Thr Arg Glu Asp Gly Gln Pro Phe
Val Ala Cys Ala Glu Cys Gly 50 55
60Phe Pro Val Cys Arg Pro Cys Tyr Glu Tyr Glu Arg Ser Glu Gly Thr65
70 75 80Gln Cys Cys Pro Gln
Cys Asn Thr Arg Tyr Lys Arg Gln Lys Gly Cys 85
90 95Pro Arg Val Glu Gly Asp Glu Glu Glu Gly Pro
Glu Met Asp Asp Phe 100 105
110Glu Asp Glu Phe Pro Ala Lys Ser Pro Lys Lys Pro His Glu Pro Val
115 120 125Ala Phe Asp Val Tyr Ser Glu
Asn Gly Glu His Pro Ala Gln Lys Trp 130 135
140Arg Thr Gly Gly Gln Thr Leu Ser Ser Phe Thr Gly Ser Val Ala
Gly145 150 155 160Lys Asp
Leu Glu Ala Glu Arg Glu Met Glu Gly Ser Met Glu Trp Lys
165 170 175Asp Arg Ile Asp Lys Trp Lys
Thr Lys Gln Glu Lys Arg Gly Lys Leu 180 185
190Asn His Asp Asp Ser Asp Asp Asp Asp Asp Lys Asn Glu Asp
Glu Tyr 195 200 205Met Leu Leu Ala
Glu Ala Arg Gln Pro Leu Trp Arg Lys Val Pro Ile 210
215 220Pro Ser Ser Met Ile Asn Pro Tyr Arg Ile Val Ile
Val Leu Arg Leu225 230 235
240Val Val Leu Cys Phe Phe Leu Lys Phe Arg Ile Thr Thr Pro Ala Thr
245 250 255Asp Ala Val Pro Leu
Trp Leu Ala Ser Val Ile Cys Glu Leu Trp Phe 260
265 270Ala Phe Ser Trp Ile Leu Asp Gln Leu Pro Lys Trp
Ala Pro Val Thr 275 280 285Arg Glu
Thr Tyr Leu Asp Arg Leu Ala Leu Arg Tyr Asp Arg Glu Gly 290
295 300Glu Ala Cys Arg Leu Ser Pro Ile Asp Phe Phe
Val Ser Thr Val Asp305 310 315
320Pro Leu Lys Glu Pro Pro Ile Ile Thr Ala Asn Thr Val Leu Ser Ile
325 330 335Leu Ala Val Asp
Tyr Pro Val Asp Arg Val Ser Cys Tyr Val Ser Asp 340
345 350Asp Gly Ala Ser Met Leu Leu Phe Asp Ala Leu
Ser Glu Thr Ala Glu 355 360 365Phe
Ala Arg Arg Trp Val Pro Phe Cys Lys Lys Phe Ala Val Glu Pro 370
375 380Arg Ala Pro Glu Phe Tyr Phe Ser Gln Lys
Ile Asp Tyr Leu Lys Asp385 390 395
400Lys Val Gln Pro Thr Phe Val Lys Glu Arg Arg Ala Met Lys Arg
Glu 405 410 415Tyr Glu Glu
Phe Lys Val Arg Ile Asn Ala Leu Val Ala Lys Ala Gln 420
425 430Lys Lys Pro Glu Glu Gly Trp Val Met Gln
Asp Gly Thr Pro Trp Pro 435 440
445Gly Asn Asn Thr Arg Asp His Pro Gly Met Ile Gln Val Tyr Leu Gly 450
455 460Asn Gln Gly Ala Leu Asp Val Glu
Gly His Glu Leu Pro Arg Leu Val465 470
475 480Tyr Val Ser Arg Glu Lys Arg Pro Gly Tyr Asn His
His Lys Lys Ala 485 490
495Gly Ala Met Asn Ala Leu Val Arg Val Ser Ala Val Leu Thr Asn Ala
500 505 510Pro Phe Ile Leu Asn Leu
Asp Cys Asp His Tyr Val Asn Asn Ser Lys 515 520
525Ala Val Arg Glu Ala Met Cys Phe Leu Met Asp Pro Gln Leu
Gly Lys 530 535 540Lys Leu Cys Tyr Val
Gln Phe Pro Gln Arg Phe Asp Gly Ile Asp Arg545 550
555 560His Asp Arg Tyr Ala Asn Arg Asn Val Val
Phe Phe Asp Ile Asn Met 565 570
575Lys Gly Leu Asp Gly Ile Gln Gly Pro Val Tyr Val Gly Thr Gly Cys
580 585 590Val Phe Asn Arg Gln
Ala Leu Tyr Gly Tyr Asp Pro Pro Arg Pro Glu 595
600 605Lys Arg Pro Lys Met Thr Cys Asp Cys Trp Pro Ser
Trp Cys Cys Cys 610 615 620Cys Cys Cys
Phe Gly Gly Gly Lys Arg Gly Lys Ala Arg Lys Asp Lys625
630 635 640Lys Gly Asp Gly Gly Glu Glu
Pro Arg Arg Gly Leu Leu Gly Phe Tyr 645
650 655Arg Lys Arg Ser Lys Lys Asp Lys Leu Gly Gly Gly
Ser Val Ala Gly 660 665 670Ser
Lys Lys Gly Gly Gly Leu Tyr Lys Lys His Gln Arg Ala Phe Glu 675
680 685Leu Glu Glu Ile Glu Glu Gly Leu Glu
Gly Tyr Asp Glu Leu Glu Arg 690 695
700Ser Ser Leu Met Ser Gln Lys Ser Phe Glu Lys Arg Phe Gly Gln Ser705
710 715 720Pro Val Phe Ile
Ala Ser Thr Leu Val Glu Asp Gly Gly Leu Pro Gln 725
730 735Gly Ala Ala Ala Asp Pro Ala Ala Leu Ile
Lys Glu Ala Ile His Val 740 745
750Ile Ser Cys Gly Tyr Glu Glu Lys Thr Glu Trp Gly Lys Glu Ile Gly
755 760 765Trp Ile Tyr Gly Ser Val Thr
Glu Asp Ile Leu Thr Gly Phe Lys Met 770 775
780His Cys Arg Gly Trp Lys Ser Val Tyr Cys Thr Pro Thr Arg Pro
Ala785 790 795 800Phe Lys
Gly Ser Ala Pro Ile Asn Leu Ser Asp Arg Leu His Gln Val
805 810 815Leu Arg Trp Ala Leu Gly Ser
Val Glu Ile Phe Met Ser Arg His Cys 820 825
830Pro Leu Arg Tyr Ala Tyr Gly Gly Arg Leu Lys Trp Leu Glu
Arg Phe 835 840 845Ala Tyr Thr Asn
Thr Ile Val Tyr Pro Phe Thr Ser Ile Pro Leu Leu 850
855 860Ala Tyr Cys Thr Ile Pro Ala Val Cys Leu Leu Thr
Gly Lys Phe Ile865 870 875
880Ile Pro Thr Leu Asn Asn Leu Ala Ser Ile Trp Phe Ile Ala Leu Phe
885 890 895Leu Ser Ile Ile Ala
Thr Ser Val Leu Glu Leu Arg Trp Ser Gly Val 900
905 910Ser Ile Glu Asp Trp Trp Arg Asn Glu Gln Phe Trp
Val Ile Gly Gly 915 920 925Val Ser
Ala His Leu Phe Ala Val Phe Gln Gly Phe Leu Lys Val Leu 930
935 940Gly Gly Val Asp Thr Ser Phe Thr Val Thr Ser
Lys Ala Ala Gly Asp945 950 955
960Glu Ala Asp Ala Phe Gly Asp Leu Tyr Leu Phe Lys Trp Thr Thr Leu
965 970 975Leu Val Pro Pro
Thr Thr Leu Ile Ile Ile Asn Met Val Gly Ile Val 980
985 990Ala Gly Val Ser Asp Ala Val Asn Asn Gly Tyr
Gly Ser Trp Gly Pro 995 1000
1005Leu Phe Gly Lys Leu Phe Phe Ser Phe Trp Val Ile Val His Leu
1010 1015 1020Tyr Pro Phe Leu Lys Gly
Leu Met Gly Arg Gln Asn Arg Thr Pro 1025 1030
1035Thr Ile Val Val Leu Trp Ser Ile Leu Leu Ala Ser Ile Phe
Ser 1040 1045 1050Leu Val Trp Val Arg
Ile Asp Pro Phe Ile Pro Lys Ala Lys Gly 1055 1060
1065Pro Ile Leu Lys Pro Cys Gly Val Glu Cys 1070
1075393231DNAZea mays 39ccacgcgtcc gggaggggcc atgatggagt
cggcggcggc ccagtcctgc gcggcgtgcg 60gggacgacgc gcgcgctgcc tgccgcgcgt
gcagctacgc gctctgcagg gcgtgcctcg 120acgaggacgc cgccgagggc cgcaccacat
gcgcgcgctg cggaggggac tacgccgcta 180tcaacccagc gcgcgccagc gagggaaccg
aggcggagga ggaggtggtg gagaaccacc 240acaccgccgg tggcctgcgt gagagggtca
ccatgggcag ccacctcaat gatcgccagg 300atgaagtaag ccacgccagg accatgagca
gcttgtcggg aattggtagt gaattgaatg 360atgaatctgg taagcccatc tggaagaaca
gggtggagag ttggaaggaa aagaagaatg 420agaagaaagc ctcggccaaa aagactgcag
ctaaagcaca gcctccgcct gtcgaagaac 480agatcatgga tgaaaaagac ttgacagatg
catatgagcc actctcccgg gtcatcccaa 540tatcaaagaa caagctcaca ccttacagag
cagtgatcat tatgcggtta attgttcttg 600ggctcttctt tcactaccgt atcaccaatc
ctgttaacag tgcctttggt ctctggatga 660catcagttat atgtgagatc tggtttggtt
tctcctggat attggatcaa ttcccgaagt 720ggtatcctat caatcgtgag acttatgttg
ataggctgat tgcacgatat ggagatggtg 780aagaatctgg gttagcacct gtagatttct
ttgtcagtac agtggatcca ttgaaagagc 840ctccactaat cactgcaaac actgtgctgt
ctattcttgc tgtggactat cccgttgaga 900agatctcatg ctatgtatct gatgatggtt
ctgctatgct cacatttgaa tcgctcgcag 960agactgcaga atatgctaga aagtgggtgc
cgttttgcaa gaagtacgcc attgagccac 1020gagctcctga gttctacttc tcacagaaaa
ttgactactt gaaggacaag atacacccat 1080cttttgtcaa ggagcgtagg gctatgaaga
gagactatga agagtacaag gtgaggataa 1140atgctttggt tgccaaggct caaaagacac
ctgatgaagg ctggatcatg caagacggta 1200caccatggcc tgggaacaat cctcgtgacc
accctggcat gatccaggtt ttcctgggtg 1260agactggtgc acgggacttt gatggaaatg
aacttcctcg gttagtgtat gtgtcaagag 1320agaaaagacc aggctaccaa caccacaaga
aggcaggggc tatgaatgct ctggtccgag 1380tgtctgctgt tctgacaaat gccccttaca
ttcttaatct tgattgtgat cactatgtta 1440acaacagcaa agctgttcgt gaagcaatgt
gcttcatgat ggaccctact gttggcagag 1500atgtctgcta tgtacaattc ccccagaggt
tcgatggcat tgatcgcagt gatcgatatg 1560ccaataggaa cgttgtgttc tttgatgtta
atatgaaagg acttgatggc ctccaaggcc 1620cagtttatgt gggaactggt tgttgtttca
ataggcaagc actttatggt tatgggcctc 1680catctctgcc cgcacttcca aagtcttcga
tttgttcctg gtgttgctgc tgctgtccca 1740agaaaaaggt tgaaagaagt gagagggaaa
tcaacagaga ctctcggcga gaagacctcg 1800agtctgccat ttttaacctt cgcgaaattg
acaactacga tgagtacgag aggtccatgc 1860tcatctctca gatgagcttc gagaagtctt
ttgggctgtc ctcggtcttt attgaatcga 1920cccttatgga gaatgggggc gtccctgaat
ctgcaaaccc atctacccta attaaagaag 1980ccattcatgt cattagctgt ggatatgaag
agaaaactga atggggaaaa gagattggct 2040ggatctatgg ttcagttaca gaggatattc
tgactgggtt taagatgcac tgccgtggct 2100ggagatccat ctactgcatg ccggtgagac
ctgcattcaa gggatcagcc ccaatcaatc 2160tttccgatcg tcttcaccaa gttctccggt
gggctcttgt ttctgtcgag atcttcttca 2220gtcggcactg cccgctgtgg tacggttacg
gtggcggccg tctgaaatgg ctccagaggc 2280tctcctacat caacaccatc gtgtacccgt
tcacttctct tcctctcgtt gcctactgtt 2340gcctgcctgc catttgcctg ctcacaggaa
agttcattat acctacgctg tccaacgctg 2400caacgatatg gtttcttggc ctcttcatgt
ccatcatcgt gacgagcgtg ttggagctgc 2460ggtggagtgg catcgggatc gaggactggt
ggcgcaacga gcagttctgg gtcatcggag 2520gcgtgtccgc gcacctgttc gccgtgttcc
agggtatcct caagatgatt gccgggctgg 2580acaccaactt cacggtcacg gcaaaggcca
cggacgacac tgagttcggg gagctgtacc 2640tgttcaagtg gacgacggtg ctgatcccgc
ccacaagcat cctggtgctg aacctggtgg 2700gcgtggtggc tgggttctcg gccgcgctca
acagcggcta cgagtcctgg ggcccgctct 2760tcggtaaggt gttcttcgcc atgtgggtga
tcatgcacct gtacccgttc ctcaagggtc 2820tcatgggccg ccagaaccgc acgccgacca
tcgtggtgct ctggtccgtc ctcctcgcct 2880ccgtcttctc cctcctgtgg gtcaagatcg
acccattcgt tggaggaacc gagaccgtca 2940acaccaacaa ctgcaacaca catctgctga
ttcaccatcg gtcagctgct gtcgtgccgc 3000ggcggacgtg tttctggtgt tgcaaacgtg
ggttgcctgc ctgatgcggg tctcctctgt 3060ctatctcgca tctgggcttt tgccccagga
tctgaagcgg gtggtgtagg ttagctttat 3120tttgcgtcca agtgttgatt gatgttgtct
gtgttatgaa aagttttggt ggtgaaacct 3180gaaatgttaa aattcggctc aattgtgaga
aaaaaaaaaa aaaaaaaaaa a 3231401007PRTZea mays 40Met Met Glu
Ser Ala Ala Ala Gln Ser Cys Ala Ala Cys Gly Asp Asp1 5
10 15Ala Arg Ala Ala Cys Arg Ala Cys Ser
Tyr Ala Leu Cys Arg Ala Cys 20 25
30Leu Asp Glu Asp Ala Ala Glu Gly Arg Thr Thr Cys Ala Arg Cys Gly
35 40 45Gly Asp Tyr Ala Ala Ile Asn
Pro Ala Arg Ala Ser Glu Gly Thr Glu 50 55
60Ala Glu Glu Glu Val Val Glu Asn His His Thr Ala Gly Gly Leu Arg65
70 75 80Glu Arg Val Thr
Met Gly Ser His Leu Asn Asp Arg Gln Asp Glu Val 85
90 95Ser His Ala Arg Thr Met Ser Ser Leu Ser
Gly Ile Gly Ser Glu Leu 100 105
110Asn Asp Glu Ser Gly Lys Pro Ile Trp Lys Asn Arg Val Glu Ser Trp
115 120 125Lys Glu Lys Lys Asn Glu Lys
Lys Ala Ser Ala Lys Lys Thr Ala Ala 130 135
140Lys Ala Gln Pro Pro Pro Val Glu Glu Gln Ile Met Asp Glu Lys
Asp145 150 155 160Leu Thr
Asp Ala Tyr Glu Pro Leu Ser Arg Val Ile Pro Ile Ser Lys
165 170 175Asn Lys Leu Thr Pro Tyr Arg
Ala Val Ile Ile Met Arg Leu Ile Val 180 185
190Leu Gly Leu Phe Phe His Tyr Arg Ile Thr Asn Pro Val Asn
Ser Ala 195 200 205Phe Gly Leu Trp
Met Thr Ser Val Ile Cys Glu Ile Trp Phe Gly Phe 210
215 220Ser Trp Ile Leu Asp Gln Phe Pro Lys Trp Tyr Pro
Ile Asn Arg Glu225 230 235
240Thr Tyr Val Asp Arg Leu Ile Ala Arg Tyr Gly Asp Gly Glu Glu Ser
245 250 255Gly Leu Ala Pro Val
Asp Phe Phe Val Ser Thr Val Asp Pro Leu Lys 260
265 270Glu Pro Pro Leu Ile Thr Ala Asn Thr Val Leu Ser
Ile Leu Ala Val 275 280 285Asp Tyr
Pro Val Glu Lys Ile Ser Cys Tyr Val Ser Asp Asp Gly Ser 290
295 300Ala Met Leu Thr Phe Glu Ser Leu Ala Glu Thr
Ala Glu Tyr Ala Arg305 310 315
320Lys Trp Val Pro Phe Cys Lys Lys Tyr Ala Ile Glu Pro Arg Ala Pro
325 330 335Glu Phe Tyr Phe
Ser Gln Lys Ile Asp Tyr Leu Lys Asp Lys Ile His 340
345 350Pro Ser Phe Val Lys Glu Arg Arg Ala Met Lys
Arg Asp Tyr Glu Glu 355 360 365Tyr
Lys Val Arg Ile Asn Ala Leu Val Ala Lys Ala Gln Lys Thr Pro 370
375 380Asp Glu Gly Trp Ile Met Gln Asp Gly Thr
Pro Trp Pro Gly Asn Asn385 390 395
400Pro Arg Asp His Pro Gly Met Ile Gln Val Phe Leu Gly Glu Thr
Gly 405 410 415Ala Arg Asp
Phe Asp Gly Asn Glu Leu Pro Arg Leu Val Tyr Val Ser 420
425 430Arg Glu Lys Arg Pro Gly Tyr Gln His His
Lys Lys Ala Gly Ala Met 435 440
445Asn Ala Leu Val Arg Val Ser Ala Val Leu Thr Asn Ala Pro Tyr Ile 450
455 460Leu Asn Leu Asp Cys Asp His Tyr
Val Asn Asn Ser Lys Ala Val Arg465 470
475 480Glu Ala Met Cys Phe Met Met Asp Pro Thr Val Gly
Arg Asp Val Cys 485 490
495Tyr Val Gln Phe Pro Gln Arg Phe Asp Gly Ile Asp Arg Ser Asp Arg
500 505 510Tyr Ala Asn Arg Asn Val
Val Phe Phe Asp Val Asn Met Lys Gly Leu 515 520
525Asp Gly Leu Gln Gly Pro Val Tyr Val Gly Thr Gly Cys Cys
Phe Asn 530 535 540Arg Gln Ala Leu Tyr
Gly Tyr Gly Pro Pro Ser Leu Pro Ala Leu Pro545 550
555 560Lys Ser Ser Ile Cys Ser Trp Cys Cys Cys
Cys Cys Pro Lys Lys Lys 565 570
575Val Glu Arg Ser Glu Arg Glu Ile Asn Arg Asp Ser Arg Arg Glu Asp
580 585 590Leu Glu Ser Ala Ile
Phe Asn Leu Arg Glu Ile Asp Asn Tyr Asp Glu 595
600 605Tyr Glu Arg Ser Met Leu Ile Ser Gln Met Ser Phe
Glu Lys Ser Phe 610 615 620Gly Leu Ser
Ser Val Phe Ile Glu Ser Thr Leu Met Glu Asn Gly Gly625
630 635 640Val Pro Glu Ser Ala Asn Pro
Ser Thr Leu Ile Lys Glu Ala Ile His 645
650 655Val Ile Ser Cys Gly Tyr Glu Glu Lys Thr Glu Trp
Gly Lys Glu Ile 660 665 670Gly
Trp Ile Tyr Gly Ser Val Thr Glu Asp Ile Leu Thr Gly Phe Lys 675
680 685Met His Cys Arg Gly Trp Arg Ser Ile
Tyr Cys Met Pro Val Arg Pro 690 695
700Ala Phe Lys Gly Ser Ala Pro Ile Asn Leu Ser Asp Arg Leu His Gln705
710 715 720Val Leu Arg Trp
Ala Leu Val Ser Val Glu Ile Phe Phe Ser Arg His 725
730 735Cys Pro Leu Trp Tyr Gly Tyr Gly Gly Gly
Arg Leu Lys Trp Leu Gln 740 745
750Arg Leu Ser Tyr Ile Asn Thr Ile Val Tyr Pro Phe Thr Ser Leu Pro
755 760 765Leu Val Ala Tyr Cys Cys Leu
Pro Ala Ile Cys Leu Leu Thr Gly Lys 770 775
780Phe Ile Ile Pro Thr Leu Ser Asn Ala Ala Thr Ile Trp Phe Leu
Gly785 790 795 800Leu Phe
Met Ser Ile Ile Val Thr Ser Val Leu Glu Leu Arg Trp Ser
805 810 815Gly Ile Gly Ile Glu Asp Trp
Trp Arg Asn Glu Gln Phe Trp Val Ile 820 825
830Gly Gly Val Ser Ala His Leu Phe Ala Val Phe Gln Gly Ile
Leu Lys 835 840 845Met Ile Ala Gly
Leu Asp Thr Asn Phe Thr Val Thr Ala Lys Ala Thr 850
855 860Asp Asp Thr Glu Phe Gly Glu Leu Tyr Leu Phe Lys
Trp Thr Thr Val865 870 875
880Leu Ile Pro Pro Thr Ser Ile Leu Val Leu Asn Leu Val Gly Val Val
885 890 895Ala Gly Phe Ser Ala
Ala Leu Asn Ser Gly Tyr Glu Ser Trp Gly Pro 900
905 910Leu Phe Gly Lys Val Phe Phe Ala Met Trp Val Ile
Met His Leu Tyr 915 920 925Pro Phe
Leu Lys Gly Leu Met Gly Arg Gln Asn Arg Thr Pro Thr Ile 930
935 940Val Val Leu Trp Ser Val Leu Leu Ala Ser Val
Phe Ser Leu Leu Trp945 950 955
960Val Lys Ile Asp Pro Phe Val Gly Gly Thr Glu Thr Val Asn Thr Asn
965 970 975Asn Cys Asn Thr
His Leu Leu Ile His His Arg Ser Ala Ala Val Val 980
985 990Pro Arg Arg Thr Cys Phe Trp Cys Cys Lys Arg
Gly Leu Pro Ala 995 1000
1005413028DNAZea mays 41cacgagttca acatcgacga cgagaatcag cagaggcagc
tggagggcaa catgcagaac 60agccagatca ccgaggcgat gctgcacggc aggatgagct
acgggagggg ccccgacgac 120ggcgacggca acaacacccc gcagatcccg cccatcatca
ccggctcccg ctccgtgccg 180gtgagcggtg agtttccgat taccaacggg tatggccacg
gcgaggtctc gtcttccctg 240cacaagcgca tccatccgta ccctgtgtct gagccaggga
gtgccaagtg ggacgagaag 300aaagaagtga gctggaagga gaggatggac gactggaagt
ccaagcaggg catcctcggc 360ggcggcgccg atcccgaaga catggacgcc gacgtggcac
tgaacgacga ggcgaggcag 420ccgctgtcga ggaaggtgtc gatcgcgtcg agcaaggtga
acccgtaccg gatggtgatc 480gtggtgcgtc tcgttgtgct cgccttcttc ctccggtacc
gtatcctgca ccccgtcccg 540gacgccatcg ggctgtggct cgtctccatc atctgcgaga
tctggttcgc catctcctgg 600atcctcgacc agttccccaa gtggttcccc atcgaccgcg
agacgtacct cgaccgcctc 660tccctcaggt acgagaggga aggggagccg tcgctgctgt
cggcggtgga cctgttcgtg 720agcacggtgg acccgctcaa ggagccgccg ctggtgaccg
ccaacaccgt gctctccatc 780ctcgccgtag actaccccgt ggacaaggtc tcctgctacg
tctccgacga cggcgcgtcg 840atgctgacgt tcgagtcgct gtcggagacg gccgagttcg
cgcgcaagtg ggtgcccttc 900tgcaagaagt tcggcatcga gccccgcgcc ccggagttct
acttctcgct caaggtcgac 960tacctcaagg acaaggtgca gcccaccttc gtgcaggagc
gccgcgccat gaagagagag 1020tatgaggagt tcaaggtccg gatcaacgcg ctggtggcca
aggccatgaa ggtgccggca 1080gaggggtgga tcatgaagga cggcacgccg tggcccggga
acaacacccg cgaccacccc 1140ggcatgatcc aggtgttcct gggccacagc ggcggccacg
acaccgaggg caacgagctg 1200ccccgcctcg tgtacgtctc ccgtgagaag cgcccgggat
tccagcacca caagaaggcc 1260ggcgccatga acgctctgat tcgcgtctcc gccgtgctga
ccaacgcgcc attcatgctc 1320aacttggact gtgatcacta catcaacaac agcaaggcca
tccgggaggc catgtgcttc 1380ctcatggacc ctcaggtcgg ccggaaggtc tgctacgttc
agttcccgca gaggttcgac 1440ggcatcgacg tgcacgaccg atacgctaac aggaacaccg
tcttcttcga catcaacatg 1500aaggggctgg acggcatcca aggcccggtg tacgtcggga
cagggtgcgt gttccggcgc 1560caggcgctct acggctacaa ccctcccaag ggacccaaga
ggcccaagat ggtgacctgc 1620gactgctgcc cgtgcttcgg ccgcaagaag cggaaacacg
ccaaggacgg gctgccggag 1680ggcaccgctg atatgggagt agatagcgac aaggagatgc
tcatgtccca catgaacttc 1740gagaagcggt tcgggcagtc cgcggcgttc gtcacgtcga
cgctgatgga ggaaggcggc 1800gtccctcctt cgtcgagccc cgccgcgctc ctcaaggagg
ccatccatgt catcagctgc 1860ggctacgagg acaagaccga ctgggggctg gagctggggt
ggatctacgg gtcgatcacg 1920gaggacatcc tgacggggtt caagatgcac tgccgcgggt
ggcgctccgt gtactgcatg 1980ccgaagcggg cggcgttcaa ggggtcggcg ccgatcaatc
tatcggaccg tctcaaccag 2040gtgctccggt gggcgctggg gtccgtcgag atcttcttca
gccggcacag ccccctgctg 2100tacggctaca agaacggcaa cctcaagtgg ctggagcgct
tcgcctacat caacaccacc 2160atctacccct tcacctcgct cccgctgctc gcctactgca
ccctccccgc cgtctgcctc 2220ctcaccggca agttcatcat gccgtcgatt agcacgttcg
ccagcctctt cttcatcgcc 2280ctcttcatgt ccatcttcgc gacgggcatc ctggagatgc
ggtggagcgg ggtgagcatc 2340gaggagtggt ggaggaacga gcagttctgg gtcatcggcg
gcgtgtccgc gcatctcttc 2400gccgtcgtgc agggcctgct caaggtcctc gccgggatcg
acaccaactt caccgtcacc 2460tccaaggcca ccggcgacga ggacgacgag ttcgccgagc
tctacgcctt caagtggacc 2520acgctcctca tcccgcccac cacgctgctc atcattaacg
tcatcggcgt cgtggccggc 2580atctccgacg ccatcaacaa cgggtaccag tcctgggggc
ccctcttcgg caagctcttc 2640ttcgccttct gggtcatcgt ccacctctac ccgttcctca
aggggctcat ggggcgccag 2700aacaggacgc ccaccgttgt tgtcatctgg tccattctgc
tggcctccat cttctccctg 2760ctctgggtca ggatcgaccc tttcatcgtc aggaccaagg
gcccggacgt caggcagtgt 2820ggcatcaatt gctgagctgt ttattaaggt tcaaaattct
ggagcttgtg catagggaga 2880aaaaaacaat ttagaaattt tgtaaggttg ttgtgtctgt
aatgttatgg tacccagaat 2940tgtcggacga ggaattgaac aaaggacaag gtttgattgt
taaatggcaa aaaaaaaaaa 3000aaaaaaaaaa aaaaaaaaaa aaaaaaaa
302842927PRTZea mays 42Met Gln Asn Ser Gln Ile Thr
Glu Ala Met Leu His Gly Arg Met Ser1 5 10
15Tyr Gly Arg Gly Pro Asp Asp Gly Asp Gly Asn Asn Thr
Pro Gln Ile 20 25 30Pro Pro
Ile Ile Thr Gly Ser Arg Ser Val Pro Val Ser Gly Glu Phe 35
40 45Pro Ile Thr Asn Gly Tyr Gly His Gly Glu
Val Ser Ser Ser Leu His 50 55 60Lys
Arg Ile His Pro Tyr Pro Val Ser Glu Pro Gly Ser Ala Lys Trp65
70 75 80Asp Glu Lys Lys Glu Val
Ser Trp Lys Glu Arg Met Asp Asp Trp Lys 85
90 95Ser Lys Gln Gly Ile Leu Gly Gly Gly Ala Asp Pro
Glu Asp Met Asp 100 105 110Ala
Asp Val Ala Leu Asn Asp Glu Ala Arg Gln Pro Leu Ser Arg Lys 115
120 125Val Ser Ile Ala Ser Ser Lys Val Asn
Pro Tyr Arg Met Val Ile Val 130 135
140Val Arg Leu Val Val Leu Ala Phe Phe Leu Arg Tyr Arg Ile Leu His145
150 155 160Pro Val Pro Asp
Ala Ile Gly Leu Trp Leu Val Ser Ile Ile Cys Glu 165
170 175Ile Trp Phe Ala Ile Ser Trp Ile Leu Asp
Gln Phe Pro Lys Trp Phe 180 185
190Pro Ile Asp Arg Glu Thr Tyr Leu Asp Arg Leu Ser Leu Arg Tyr Glu
195 200 205Arg Glu Gly Glu Pro Ser Leu
Leu Ser Ala Val Asp Leu Phe Val Ser 210 215
220Thr Val Asp Pro Leu Lys Glu Pro Pro Leu Val Thr Ala Asn Thr
Val225 230 235 240Leu Ser
Ile Leu Ala Val Asp Tyr Pro Val Asp Lys Val Ser Cys Tyr
245 250 255Val Ser Asp Asp Gly Ala Ser
Met Leu Thr Phe Glu Ser Leu Ser Glu 260 265
270Thr Ala Glu Phe Ala Arg Lys Trp Val Pro Phe Cys Lys Lys
Phe Gly 275 280 285Ile Glu Pro Arg
Ala Pro Glu Phe Tyr Phe Ser Leu Lys Val Asp Tyr 290
295 300Leu Lys Asp Lys Val Gln Pro Thr Phe Val Gln Glu
Arg Arg Ala Met305 310 315
320Lys Arg Glu Tyr Glu Glu Phe Lys Val Arg Ile Asn Ala Leu Val Ala
325 330 335Lys Ala Met Lys Val
Pro Ala Glu Gly Trp Ile Met Lys Asp Gly Thr 340
345 350Pro Trp Pro Gly Asn Asn Thr Arg Asp His Pro Gly
Met Ile Gln Val 355 360 365Phe Leu
Gly His Ser Gly Gly His Asp Thr Glu Gly Asn Glu Leu Pro 370
375 380Arg Leu Val Tyr Val Ser Arg Glu Lys Arg Pro
Gly Phe Gln His His385 390 395
400Lys Lys Ala Gly Ala Met Asn Ala Leu Ile Arg Val Ser Ala Val Leu
405 410 415Thr Asn Ala Pro
Phe Met Leu Asn Leu Asp Cys Asp His Tyr Ile Asn 420
425 430Asn Ser Lys Ala Ile Arg Glu Ala Met Cys Phe
Leu Met Asp Pro Gln 435 440 445Val
Gly Arg Lys Val Cys Tyr Val Gln Phe Pro Gln Arg Phe Asp Gly 450
455 460Ile Asp Val His Asp Arg Tyr Ala Asn Arg
Asn Thr Val Phe Phe Asp465 470 475
480Ile Asn Met Lys Gly Leu Asp Gly Ile Gln Gly Pro Val Tyr Val
Gly 485 490 495Thr Gly Cys
Val Phe Arg Arg Gln Ala Leu Tyr Gly Tyr Asn Pro Pro 500
505 510Lys Gly Pro Lys Arg Pro Lys Met Val Thr
Cys Asp Cys Cys Pro Cys 515 520
525Phe Gly Arg Lys Lys Arg Lys His Ala Lys Asp Gly Leu Pro Glu Gly 530
535 540Thr Ala Asp Met Gly Val Asp Ser
Asp Lys Glu Met Leu Met Ser His545 550
555 560Met Asn Phe Glu Lys Arg Phe Gly Gln Ser Ala Ala
Phe Val Thr Ser 565 570
575Thr Leu Met Glu Glu Gly Gly Val Pro Pro Ser Ser Ser Pro Ala Ala
580 585 590Leu Leu Lys Glu Ala Ile
His Val Ile Ser Cys Gly Tyr Glu Asp Lys 595 600
605Thr Asp Trp Gly Leu Glu Leu Gly Trp Ile Tyr Gly Ser Ile
Thr Glu 610 615 620Asp Ile Leu Thr Gly
Phe Lys Met His Cys Arg Gly Trp Arg Ser Val625 630
635 640Tyr Cys Met Pro Lys Arg Ala Ala Phe Lys
Gly Ser Ala Pro Ile Asn 645 650
655Leu Ser Asp Arg Leu Asn Gln Val Leu Arg Trp Ala Leu Gly Ser Val
660 665 670Glu Ile Phe Phe Ser
Arg His Ser Pro Leu Leu Tyr Gly Tyr Lys Asn 675
680 685Gly Asn Leu Lys Trp Leu Glu Arg Phe Ala Tyr Ile
Asn Thr Thr Ile 690 695 700Tyr Pro Phe
Thr Ser Leu Pro Leu Leu Ala Tyr Cys Thr Leu Pro Ala705
710 715 720Val Cys Leu Leu Thr Gly Lys
Phe Ile Met Pro Ser Ile Ser Thr Phe 725
730 735Ala Ser Leu Phe Phe Ile Ala Leu Phe Met Ser Ile
Phe Ala Thr Gly 740 745 750Ile
Leu Glu Met Arg Trp Ser Gly Val Ser Ile Glu Glu Trp Trp Arg 755
760 765Asn Glu Gln Phe Trp Val Ile Gly Gly
Val Ser Ala His Leu Phe Ala 770 775
780Val Val Gln Gly Leu Leu Lys Val Leu Ala Gly Ile Asp Thr Asn Phe785
790 795 800Thr Val Thr Ser
Lys Ala Thr Gly Asp Glu Asp Asp Glu Phe Ala Glu 805
810 815Leu Tyr Ala Phe Lys Trp Thr Thr Leu Leu
Ile Pro Pro Thr Thr Leu 820 825
830Leu Ile Ile Asn Val Ile Gly Val Val Ala Gly Ile Ser Asp Ala Ile
835 840 845Asn Asn Gly Tyr Gln Ser Trp
Gly Pro Leu Phe Gly Lys Leu Phe Phe 850 855
860Ala Phe Trp Val Ile Val His Leu Tyr Pro Phe Leu Lys Gly Leu
Met865 870 875 880Gly Arg
Gln Asn Arg Thr Pro Thr Val Val Val Ile Trp Ser Ile Leu
885 890 895Leu Ala Ser Ile Phe Ser Leu
Leu Trp Val Arg Ile Asp Pro Phe Ile 900 905
910Val Arg Thr Lys Gly Pro Asp Val Arg Gln Cys Gly Ile Asn
Cys 915 920 925
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