Patent ID: 7910345
Filing Date: 2011-03-22
Classification: C12N,C12P

Abstract:
1. 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 comprising at least 90% sequence identity with SEQ ID NO: 18, wherein the O—RS further comprises a Cys, Thr or Gly at a position corresponding to position 159 and comprises Arg or Leu at a position corresponding to position 162 compared to wild type tyrosyl tRNA synthetase from 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.