RECOMBINANT PLASMID ENCODING FUSED PROTEIN AS PRECURSOR OF HUMAN INSULIN (VARIANTS), STRAIN OF BACTERIUM ESCHERICHIA COLI - PRODUCER OF FUSED PROTEIN AS PRECURSOR OF HUMAN INSULIN (VARIANTS), METHOD OF HUMAN INSULIN PREPARING

摘要

FIELD: biotechnology, molecular biology, genetic engineering. SUBSTANCE: invention relates to development of new recombinant plasmids expressing fused protein where amino acid sequences of lider peptide that is IgG-binding domain of protein A and human proinsulin are bound by two arginine residues (plasmid pRRproins) or lysine and arginine residues (plasmid pRRproins). Human insulin is prepared by a method that involves culturing the strain E. coli JM-109-pRRproins carrying plasmid pRRproins or the strain E. coli JM-109-pRRproins carrying plasmid pRRproins, isolation of inclusion bodies, solubilization, sulfitolysis and renaturation of fused protein by ion-exchange chromatography method, its cleavage with trypsin-like and carboxypeptidase B-like enzymes and insulin isolation. EFFECT: simplified technological process of human insulin preparing, increased yield of insulin. 6 cl, 4 dwg, 7 ex