Patent ID: 8603775
Filing Date: 2013-12-10
Classification: C07K,C12P

Abstract:
1. A method of synthesizing a protein having intermolecular or intramolecular disulfide bonds, comprising using a reaction system comprising a), b), c), and d) below: a) at least one template nucleic acid encoding a protein of interest, b) a protein synthesis reaction reagent comprising 1) synthesized or isolated ribosomes, initiation factors, elongation factors, termination factors, aminoacyl-tRNA synthetases, methionyl tRNA transformylases, tRNAs, amino acids, ribonucleoside triphospates, 10-formyl 5,6,7,8-tetrahyrofolic acid (FD), salts and water or 2) synthesized or isolated ribosomes, initiation factors, elongation factors, aminoacyl-tRNA synthetases, methionyl tRNA transformylases, tRNAs, amino acids, ribonucleoside triphosphates, 10-formyl 5,6,7,8-tetrahydrofolic acid (FD), salts, and water, c) at least one purified oxidoreductase, and d) an effective amount of a purified dithiothreitol and at least one purified redox reagent, adding c) the at least one purified oxidoreductase for catalyzing oxidation-reduction of disulfide bonds d) the purified dithiothreitol and the at least one redox reagent for conditioning a redox state of disulfide bonds to b) the protein synthesis reaction reagent; thereafter adding the template nucleic acid a) to the protein synthesis reaction reagent b), thereby causing a reaction to synthesize a protein encoded by the template nucleic acid; and allowing the synthesized protein to form intermolecular or intramolecular disulfide bonds.