FIELD: biotechnology.
SUBSTANCE: invention relates to biotechnology, specifically to recombinant production of therapeutic proteins, and can be used to produce recombinant human proinsulin aspart. Recombinant proinsulin is obtained in a hybrid polypeptide, in which the human HGS protein fragment, the tyrosine kinase substrate, is linked through peptide linker with sequence GlyHis6GlySerArg with amino acid sequence of proinsulin of human aspart, with arginine as site of proteolysis between C-peptide and chain A. For this purpose, recombinant plasmid DNA pF646 and strain-producer Escherichia coli BL21/pF646 are used.
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 proportion of insulin aspart in the hybrid protein to 49_5 % and improve the purification efficiency of the end product by eliminating the closely related Arg0-A-aspart admixture.
3 cl, 6 dwg, 6 ex
