Plant aminoacyl-tRNA synthetase
First Claim
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1. An isolated nucleic acid fragment encoding an isoleucyl-tRNA synthase comprising a member selected from the group consisting of:
- (a) an isolated nucleic acid fragment encoding an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO;
2, 4, 6 and 8;
(b) an isolated nucleic acid fragment that is complementary to (a).
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Abstract
This invention relates to an isolated nucleic acid fragment encoding an aminoacyl-tRNA synthetase. The invention also relates to the construction of a chimeric gene encoding all or a portion of the aminoacyl-tRNA synthetase, in sense or antisense orientation, wherein expression of the chimeric gene results in production of altered levels of the aminoacyl-tRNA synthetase in a transformed host cell.
65 Citations
30 Claims
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1. An isolated nucleic acid fragment encoding an isoleucyl-tRNA synthase comprising a member selected from the group consisting of:
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(a) an isolated nucleic acid fragment encoding an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in SEQ ID NO;
2, 4, 6 and 8;
(b) an isolated nucleic acid fragment that is complementary to (a). - View Dependent Claims (2, 3, 4, 5, 26)
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6. An isoleucyl-tRNA synthase polypeptide comprising all or a substantial portion of the amino acid sequence set forth in a member selected from the group consisting of SEQ ID NO:
- 2,4,6and 8.
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7. An isolated nucleic acid fragment encoding a lysyl-tRNA synthetase comprising a member selected from the group consisting of:
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(a) an isolated nucleic acid fragment encoding an amino acid sequence that is at least 90% identical to the amino acid sequence set forth in a member selected from the group consisting of SEQ ID NO;
10, 12, 14, and 16;
(b) an isolated nucleic acid fragment that is complementary to (a). - View Dependent Claims (8, 9, 10, 11, 14, 15, 16, 17, 20, 21, 22, 23, 25, 28, 29)
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12. A lysyl-tRNA synthetase polypeptide comprising all or a substantial portion of the amino acid sequence set forth in a member selected from the group consisting of SEQ ID NO:
- 10, 12, 14 and 16.
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13. An isolated nucleic acid fragment encoding a phenylalanyl-tRNA synthetase comprising a member selected from the group consisting of:
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(a) an isolated nucleic acid fragment encoding an amino acid sequence that is at least 80% identical to the amino acid sequence set forth in a member selected from the group consisting of SEQ ID NO;
18, 20, 22 and 24;
(b) an isolated nucleic acid fragment that is complementary to (a).
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18. A phenylalanyl-tRNA synthetase polypeptide comprising all or a substantial portion of the amino acid sequence set forth in a member selected from the group consisting of SEQ ID NO:
- 18, 20, 22 and 24.
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19. An isolated nucleic acid fragment encoding a prolyl-tRNA synthetase comprising a member selected from the group consisting of:
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(a) an isolated nucleic acid fragment encoding an amino acid sequence of at least 152 contiguous amino acids and that is at least 80% identical to the amino acid sequence set forth in a member selected from the group consisting of SEQ ID NO;
26, 28 and 30;
(b) an isolated nucleic acid fragment that is complementary to (a).
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24. A prolyl-tRNA synthetase polypeptide comprising all or a substantial portion of the amino acid sequence set forth in a member selected from the group consisting of SEQ ID NO:
- 26, 28 and 30.
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27. A method of obtaining a nucleic acid fragment encoding a substantial portion of an amino acid sequence encoding an aminoacyl-tRNA synthetase comprising:
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(a) synthesizing an oligonucleotide primer corresponding to a portion of the sequence set forth in any of SEQ ID NOs;
1, 3, 5, 7,9, 11, 13, 15, 17, 19, 21, 23, 25, 27 and 29;
and(b) amplifying a cDNA insert present in a cloning vector using the oligonucleotide primer of step (a) and a primer representing sequences of the cloning vector wherein the amplified nucleic acid fragment encodes a substantial portion of an amino acid sequence encoding an aminoacyl-tRNA synthetase.
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30. A method for evaluating at least one compound for its ability to inhibit the activity of an aminoacyl-tRNA synthetase, the method comprising the steps of:
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(a) transforming a host cell with a chimeric gene comprising a nucleic acid fragment encoding an aminoacyl-tRNA synthetase, operably linked to suitable regulatory sequences;
(b) growing the transformed host cell under conditions that are suitable for expression of the chimeric gene wherein expression of the chimeric gene results in production of the aminoacyl-tRNA synthetase encoded by the operably linked nucleic acid fragment in the transformed host cell;
(c) optionally purifying the aminoacyl-tRNA synthetase expressed by the transformed host cell;
(d) treating the aminoacyl-tRNA synthetase with a compound to be tested; and
(e) comparing the activity of the aminoacyl-tRNA synthetase that has been treated with a test compound to the activity of an untreated aminoacyl-tRNA synthetase, thereby selecting compounds with potential for inhibitory activity.
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Specification