FIELD: chemistry; biochemistry.
SUBSTANCE: present invention pertains to genetic engineering, specifically to microbial production of recombinant proteins, and can be used in biotechnology, medicine and veterinary medicine. Designed is a recombinant plasmid DNA pET-KSI-Buf2 with size 4066 "п.о.", coding hybrid protein KSI-Buf2, containing antimicrobial peptide buforin-2. Through transformation of cells of the parent strain of the Escherichia coli BL21(DE3) bacteria of recombinant plasmid DNA pET-KSI-Buf2, the strain Escherichia coli BL21(DE3)/pET-KSI-Buf2 - producer of hybrid protein, is obtained, containing antimicrobial peptide buforin-2. The Escherichia coli BL21(DE3)/pET-KSI-Buf2 strain is then cultured, with subsequent cell disintegration, separation and washing of inclusion bodies, containing hybrid protein KSI-Buf2, solubilisation of the hybrid protein and its splitting using bromine cyan in an acid medium on a methionine residue, dissolving of the target peptide and cleaning it using high performance liquid chromatography method.
EFFECT: obtaining identical natural buforin-2 with high output, using simple technology.
3 cl, 5 dwg, 1 tbl, 4 ex
