FIELD: biotechnology.
SUBSTANCE: invention relates to biotechnology, specifically to recombinant production of therapeutic proteins, and can be used to produce human recombinant B30-desthreonine-insulin. Recombinant B30-desthreonine-insulin is obtained in a hybrid polypeptide, in which the human HGS protein fragment sequence, tyrosine kinase substrate is connected through a peptide linker with the sequence GlyHis6GlySerArg with amino acid sequence B30-desthreonine-insulin human, with arginine as a site of proteolysis between C-peptide and chain A. For this purpose recombinant plasmid DNA pF882 and strain-producer Escherichia coli BL21/pF882.
EFFECT: invention provides synthesis of a hybrid polypeptide with an expression level of not less than 4 % of the crude weight of the cell biomass, enables to increase the portion of B30-desthreonine insulin in the hybrid protein to 49_5 % and increases efficiency of purification of end product due to elimination of closely related admixture of Arg0-A-(B30-desthreonine-insulin).
3 cl, 6 dwg, 5 ex
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Year |
Author |
Number |
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