FIELD: biotechnology; pharmacology.
SUBSTANCE: invention relates to biotechnology and pharmacology, namely to a method of producing recombinant protein OMP25d-OMP19-OMP10His, suitable for use in the development and production of vaccines. Proposed method involves: constructing a gene for protein OMP25d-OMP19-OMP10His containing genes of three brucellosis proteins separated by two linkers; creation of recombinant plasmid pET32b sdWP_002965367.1-sdWP_002964998.1-sdWP_002966502.1-His, 6871 bp, carrying a sequence coding chimeric protein OMP25d-OMP19-OMP10His, 1524 bp, fused with a synthetic sequence coding six histidines; transformation of E. coli BL21(DE3) cells with said recombinant plasmid to obtain a strain E. coli BL21(DE3) / pET32b sdWP_002965367.1-sdWP_002964998.1-sdWP_002966502.1-His; cultivation of this strain in a medium with an antibiotic under IPTG control at 37 °C for 4 hours; obtaining a fraction of inclusion bodies by ultrasound treatment of cells, followed by centrifugation; purification by metal-chelate chromatography in denaturing conditions.
EFFECT: invention provides production of homogeneous recombinant protein with purity of 98%, bacterial endotoxin content of not more than 5 IU/mg and output of 0.2 g per litre of bacterial culture.
1 cl, 4 dwg, 2 tbl, 6 ex
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