PRODUCTION OF RECOMBINANT TYPE HUMAN ANGIOTENSINOGEN

摘要

PURPOSE:To enable obtaining of the subject substance in a large amount and good yield by inserting a specific vector into a cell according to a calcium phosphate method and then culturing the resultant cell in a culture medium for growing the cells. CONSTITUTION:A recombinant type human angiotensinogen cDNA is integrated into cleaved sites of an expression vector in an animal cultured cell with restriction enzymes BamHI/EcoRV to provide an expression vector pSVHagD (A), which is then inserted into an animal cell (dhfr-deficient strain, DXB-11) of a Chinese hamster ovary (CHO) cell, etc., in the presence of 0.5-1.0M calcium phosphate to afford an inserted cell (D). The resultant cell (B) is subsequently cultured in a culture medium prepared by slowly adding aminopterin so as to provide 50-1000nM concentration thereof at about 37 deg.C for a desired number of days to afford a culture supernatant (C). The ingredient (C) is then purified by a salting out, affinity chromatography, isoelectric focusing method, etc., to produce a recombinant type human angiotensinogen.