RECOMBINANT PLASMID DNA pBK415 CODING POLYPEPTIDE OF RECOMBINANT TISSULAR ACTIVATOR OF PLASMINOGEN OF HUMAN BEING, CELL LINE Cricetulus griseus CHO 1F8-PRODUCER OF RECOMBINANT TISSULAR ACTIVATOR OF PLASMINOGEN OF HUMAN BEING, AND METHOD FOR OBTAINING AND SEPARATION OF POLYPEPTIDE HAVING ACTIVITY OF TISSULAR ACTIVATOR OF PLASMINOGEN

摘要

FIELD: biotechnologies.;SUBSTANCE: recombinant plasmid DNA pBK415 coding polypeptide with sequence of tissular activator of human plasminogen, also including MAR - binding area to nuclear matrix of lysozyme gene of birds, virus transmission enhancer CMV, internal translation initiation site IRES of encephalomyocarditis virus, gene DHFR of a mouse, a polyadenylation signal of virus SV40, gene of aminoglycoside-3'-phosphotransferase providing stability to geneticin (Neo) and a cassette for expression in bacteria cells of gene of β-lactamase providing stability to ampicillin, cells of line Cricetulus griseus CHO DHFR(-) are obtained so that there produced is cell line Cricetulus griseus CHO 1F8 producing recombinant protein of tissular activator of plasminogen with highly stable yield at the level of up to 190 mg/l. Cultivation of cells-producers is performed under perfusion conditions in presence of a mixture consisting of additive CHO Bioreactor supplement and sodium butyrate or dimethylsulphoxide with further separation of a target product.;EFFECT: improvement of the method.;5 cl, 5 dwg, 3 tbl, 8 ex