Patent ID: 7524647
Filing Date: 2009-04-28
Classification: C12N,C12P

Abstract:
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 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.