Compositions of Aminoacyl-tRNA Synthetase and Uses Thereof
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Abstract
Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNA'"'"'s, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNA'"'"'s/synthetases are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins using these orthogonal pairs.
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Citations
45 Claims
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1-18. -18. (canceled)
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19. A cell comprising a translation system, wherein the translation system comprises an aminoacyl tRNA synthetase (RS) having the polynucleotide sequence selected from the group consisting of SEQ ID NO:
- 5, and a complementary polynucleotide sequence of SEQ ID NO;
5. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
- 5, and a complementary polynucleotide sequence of SEQ ID NO;
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31-35. -35. (canceled)
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36. A vector comprising a polynucleotide encoding an RS having polynucleotide sequence encoding the amino acid sequence of SEQ ID NO:
- 5, and a complementary polynucleotide sequence thereof.
- View Dependent Claims (37, 38, 39)
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40. A method of producing a polypeptide in a cell with a selected amino acid at a specified position, the method comprising:
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growing, in an appropriate medium, the cell, where the cell comprises a nucleic acid that comprises at least one selector codon and encodes a polypeptide; and
,providing the selected amino acid; wherein the cell further comprises; an orthogonal RS (O-RS) that functions in the cell wherein said RS has an amino acid sequence selected from the group consisting of SEQ ID NO;
5, and a complementary polynucleotide sequence thereof, and,an orthogonal tRNA (O-tRNA), that recognizes the selector codon wherein said O-RS aminoacylates the O-tRNA with the selected amino acid. - View Dependent Claims (44)
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41. The method of claim 40 wherein said selected amino acid is para-acetyl phenylalanine.
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42-41. -41. (canceled)
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45. A polynucleotide having a polynucleotide sequence encoding the amino acid sequence of SEQ ID NO:
- 5, and a complementary polynucleotide sequence thereof.
Specification