RECOMBINANT PLASMID DNA pAK380 CODING POLYPEPTIDE OF RECOMBINANT FACTOR IX OF HUMAN BLOOD COAGULABILITY, LINE OF CELLS Cricetulus griseus CHO, 1E6-PRODUCER OF RECOMBINANT FACTOR IX OF HUMAN BLOOD COAGULABILITY, AND METHOD FOR OBTAINING POLYPEPTIDE HAVING ACTIVITY OF RECOMBINANT FACTOR IX Russian patent published in 2013 - IPC C12N15/11 C12N15/63 C12N5/10 C12P21/02 

FIELD: biotechnologies.

SUBSTANCE: invention can be used for obtaining recombinant blood coagulability factor IX of human being (hFIX). Recombinant plasmid DNA pAK380 containing gene of protein rhFIX, MAR - binding area to nuclear matrix of lysozyme gene of birds, virus transcription enhancer CMV and an internal translation initiation site IRES of encephalomyocarditis virus, gene DHFR of a mouse, a polyadenylation signal of virus SV40, gene of aminoglycoside-3'-phosphotransferase for stability to geneticin (Neo), a cassette for expression in bacteria cells of gene β-lactamase for stability to ampicillin, is used for obtaining recombinant factor hFIX in cells of line Cricetulus griseus CHO 1E6. By transformation of cell line C. griseus CHO DHFR - recombinant plasmid DNA pAK380 there obtained is cell line C. griseus CHO 1E6 producing recombinant hFIX with stable high yield at the level of 50 mg/l/24 h. After cultivation of cells-producers there extracted is hFIX by pseudoaffine chromatography on Q Sepharose with elution of 10mM CaCl2; then, on Heparin-Sepharose FF with elution of 600 mM NaCl, and chromatography on hydroxyapatite of type I with elution of 600 mM K3PO3 and chromatography on Source 30Q with elution of 600 mM with ammonium acetate.

EFFECT: improvement of the method.

4 cl, 5 dwg, 7 ex, 3 tbl