| US 7,524,647 B2 | ||
| Site specific incorporation of keto amino acids into proteins | ||
| Peter G. Schultz, La Jolla, Calif. (US); and Lei Wang, San Diego, Calif. (US) | ||
| Assigned to The Scripps Research Institute, La Jolla, Calif. (US) | ||
| Filed on Oct. 26, 2007, as Appl. No. 11/978,160. | ||
| Application 11/978160 is a continuation of application No. 10/530421, previously published as PCT/US03/32576, filed on Oct. 15, 2003. | ||
| Claims priority of provisional application 60/419265, filed on Oct. 16, 2002. | ||
| Claims priority of provisional application 60/420990, filed on Oct. 23, 2002. | ||
| Prior Publication US 2008/0176284 A1, Jul. 24, 2008 | ||
| This patent is subject to a terminal disclaimer. | ||
| Int. Cl. C12P 21/06 (2006.01); C12N 9/22 (2006.01); C12N 15/87 (2006.01) | ||
| U.S. Cl. 435—69.1 [435/91.2; 435/483; 435/440; 435/419; 435/455; 530/350] | 7 Claims |
| 1. 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 keto amino acid is a derivative of a tyrosine or a phenylalanine 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) comprising an amino acid sequence selected from SEQ ID NO: 18 or a conservative
variation thereof comprising at least 90% identity with SEQ ID NO: 18;
wherein the ORS further comprises as Ala or Leu at a position corresponding to position 32 and comprises Gly at a position
corresponding to position 158 compared to wild type tyrosy 1 tRNA synthetase from Methanococcus jannaschii; and
incorporating the keto amino acid into the specified position in the protein during translation of the nucleic acid with the
at least one selector codon, thereby producing the protein.
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