STRAIN OF BACTERIA ESCHERICHIA COLI - PRODUCER heat shock protein 70 AND METHOD FOR THE PREPARATION OF HUMAN heat shock proteins

摘要

1. A strain of bacteria Escherichia coli BB 1553 28.11.2012 pMSHsp70 deposited in the Russian National Collection of Industrial Microorganisms (VKPM) State-Federal State Unitary Enterprise NIIGenetika under №V-11388 - producing HSP 70.2. A method of producing recombinant human heat shock protein, comprising culturing cells of the recombinant producer strain Escherichia coli, isolation of the bacterial biomass of the recombinant protein and its purification from impurities bacterial proteins using chromatography, characterized in that as the producer strain was cultured strain Escherichia coli BB 1553, transformed with the vector plasmid pMSHsp70, protein excretion is performed repeatedly freeze-thawing of cells and treating the resulting slurry consecutively flax solution of PS-DNA and magnesium chloride solution, followed by centrifugation and the protein containing solution supernatants chromatographic purification, which is carried out sequentially on an ion exchange sorbent DEAE-Sepharose equilibrated with a buffer solution at pH 7.5 and affinity chromatography on ATP-agarose, followed by a solution obtained after affinity chromatography was purified from the gel by incubating lipopolysaccharides Polymyxin B Agarozoy.3. A method according to claim 2, characterized in that the purification on an ion exchange sorbent DEAE-Sepharose was carried out by pre-equilibrated with five column volumes of buffer containing 20 mM Tris-HCl, 20 mM NaCl, 0,1 mM EDTA at pH 7.5 and two volumes of the same buffer containing 0.1% Triton X-100 and 0.2 mM phenyl methyl sulfonyl fluoride, and 1.5 volumes of elution 0.35 M NaCl in buffer rastvore.4. A method according to claim 2, characterized in that the purification on a column of ATP agarose