Patent ID: 7811801
Filing Date: 2010-10-12
Classification: C12N,C12P

Abstract:
1. A method of producing a protein in a cell with a redox active 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 redox active 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 the redox active amino acid; and, incorporating the redox active 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; wherein the O-RS comprises an amino acid sequence comprising SEQ ID NO.: 1 or a conservative variation thereof comprising at least 90% identity to SEQ ID NO: 1, and wherein the O-RS further comprises a Leu amino acid in a position of the O-RS corresponding to Tyr32 of SEQ ID NO: 4, a Ser amino acid residue in a position of the O-RS corresponding to Ala67 of SEQ ID NO: 4, an Asn amino acid residue in a position of the O-RS corresponding to His70 of SEQ ID NO: 4 and a Gln residue in a position of the O-RS corresponding to Ala167 of SEQ ID NO: 4 and SEQ ID NO: 4 is the wild type sequence.