FIELD: biotechnology.
SUBSTANCE: group of inventions refers to biotechnology and concerns production of recombinant barnase. Recombinant plasmid pET-STII-Ba is proposed that provides synthesis in Escherichia coli cells of barnase having the amino acid sequence of SEQ ID No. 3. This plasmid was obtained by cloning a recombinant DNA fragment with nucleotide sequence of SEQ ID No. 1 to the plasmid vector pET39b at the NdeI/HindIII restriction sites. Recombinant strain of Escherichia coli BL21(DE3)/pET-STII-Ba, containing recombinant plasmid pET-STII-Ba and producing barnase. Method for producing barnase with amino acid sequence of SEQ ID No. 3, from biomass strain Escherichia coli BL21(DE3)/pET-STII-Ba includes the isolation of protein from the periplasm of cells grown at a reduced temperature of 25 ºC, and subsequent purification by chromatography on phosphocellulose and a cation exchanger.
EFFECT: group of inventions allows to increase and stabilize the production of recombinant barnase upon accumulation of the latter in the periplasm of cells grown at a reduced temperature 25 °C.
3 cl, 4 dwg, 6 ex
