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US 2009/0215117 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 p-acetyl-L-phenylalanine 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 p-acetyl-L-phenylalanine;
- 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 the p-acetyl-L-phenylalanine;
- and wherein the O-RS comprises an amino acid sequence of SEQ ID NO:
- 18 or a conservative variation thereof comprising at least 90% sequence identity with SEQ ID NO.:
- 18;
- and incorporating the p-acetyl-L-phenylalanine into the specified position in the protein during translation of the nucleic acid with the at least one selector codon, thereby producing the protein.
8 more elements
(canceled)
Dependencies are inferred from the claim text.