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US 2008/0176284 A1Published application
Site specific incorporation of keto amino acids into proteins
AssigneeThe Scripps Research Institute
InventorsPeter G. Schultz, Lei Wang
Life of this patent
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Abstract
Compositions and methods of producing components of protein biosynthetic machinery that include orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases, which incorporate keto amino acids into proteins are provided. Methods for identifying these orthogonal pairs are also provided along with methods of producing proteins with keto amino acids using these orthogonal pairs.
Claims7
1-22. (canceled)
A method of producing a protein in a cell with a keto amino acid at a specified position, the method comprising:
- growing, in an appropriate medium, the cell, where the cell comprises a nucleic acid that comprises at least one selector codon and encodes a protein;
- and,
- providing the keto amino acid;
- wherein the cell further comprises:
- an orthogonal tRNA (O-tRNA) that functions in the cell and recognizes the selector codon;
- and, an orthogonal aminoacyl-tRNA synthetase (O-RS) that preferentially aminoacylates the O-tRNA with an efficiency of at least 50% of the efficiency of a polypeptide comprising an amino acid sequence of SEQ ID NO.:
- 18 with the keto amino acid;
- and
- incorporating the keto amino acid into the specified position in the protein during translation of the nucleic acid with the at least two selector codons, wherein said selector codons are selected from a group consisting of UGA, UAG, and UAA stop codons, thereby producing the protein.
6 more elements
Dependencies are inferred from the claim text.