RECOMBINANT PLASMID DNA PPINS16 ENCODING FUSED POLYPEPTIDE CONTAINING HUMAN PROINSULIN, RECOMBINANT PLASMID DNA PPINS25 ENCODING FUSED POLYPEPTIDE CONTAINING HUMAN PROINSULIN AND STRAIN OF BACTERIUM ESCHERICHIA COLI - PRODUCER OF FUSED POLYPEPTIDE CONTAINING HUMAN PROINSULIN Russian patent published in 1999 - IPC

FIELD: molecular biology, biotechnology, genetic engineering. SUBSTANCE: recombinant plasmid DNA pPINS16 has an artificial gene encoding fused polypeptide consisting of a single IgG-binding domain of protein A from S. aureus, peptide linker His₆-Gly-Ser-Arg and human proinsulin, tandem of E. coli tryptophan operons tandem and a synthetic enhancer of gene 10 translation from bacteriophage T7. Recombinant plasmid DNA pPINS16 provides the constitutive biosynthesis of fused polypeptide in E. coli SG20050 cells at the level 25-30% of total cellular protein. Recombinant plasmid DNA pPINS25 has an artificial gene encoding fused polypeptide consisting of a single IgG-binding domain of protein A from S. aureus, peptide linker His₆-Gly- -Ser-Arg and human proinsulin, hybrid trc-promoter, tandem of two terminators of E. coli ribosomal operon and mutant gene encoding thermosensitive lac-repressor. The strain Escherichia coli SG20050/pPINS25 - producer of fused polypeptide containing human proinsulin. Invention ensures to induce the biosynthesis of the end fused polypeptide by shift of the culturing temperature of the strain-producer from 30 C to 42 C. The level of biosynthesis is 25% of the total cellular protein, not less. Invention can be used for producing recombinant human proinsulin. EFFECT: improved method of proinsulin producing, increased yield. 4 cl, 3 dwg, 3 ex