Plant amino acyl-tRNA synthetase
OwnerOmolayo O. Famodu, Carl R. Simmonsinventors
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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.
Claims30
An isolated nucleic acid fragment encoding an aspartyl-tRNA synthetase 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 a member selected from the group consisting of SEQ ID NO:
- 2, 4, 6 and 8;
- (b) an isolated nucleic acid fragment that is complementary to (a).
An aspartyl-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:
- 2., 4, 6 and 8
An isolated nucleic acid fragment encoding a cysteinyl-tRNA synthetase 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 a member selected from the group consisting of SEQ ID NO:
- 10, 12 and 14;
- (b) an isolated nucleic acid fragment that is complementary to (a).
A cysteinyl-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 and 14.
An isolated nucleic acid fragment encoding a tryptophanyl-tRNA synthetase 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 a member selected from the group consisting of SEQ ID NO:
- 16, 18 and 20;
- (b) an isolated nucleic acid fragment that is complementary to (a).
A tryptophanyl-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:
- 16, 18 and 20.
An isolated nucleic acid fragment encoding a tyrosyl-tRNA synthetase 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:
- 22;
- (b) an isolated nucleic acid fragment that is complementary to (a).
A tyrosyl-tRNA synthetase polypeptide comprising all or a substantial portion of the amino acid sequence set forth in SEQ ID NO:
- 22.
A method of obtaining a nucleic acid fragment encoding a substantial portion of an amino acid sequence encoding an aminoacyl-tRNA synthetase comprising:
- (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 and 21;
- 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.
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:
- (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.
Dependencies are inferred from the claim text.