After the eukaryotic source containing disulfide bond obtained by the activation method of gene technology heterologous protein,It is expressed in prokaryotes

摘要

Method includes cytolysis for activating non-glycosylated tissue plasminogen activator (t-PA) to show in prokaryotic cell at it; Under dissolution denaturation reduction situation, reactivation is under oxidative conditions in the glutathione (G5H G55G) that there is reduction and oxidation. Feature is first is that final stage is in (the counties 9. pH 9-125-11) with G5H and G55G concn. 0.1-20, county, 0.2-10 millimoles and 0.01-3, prefix 0.5-1, millimole, and it is respectively provided with the concentration of non-denaturing denaturant. Especially this method is to express E for t-PA. Coliform and P. Putidaredoxin. Denaturant is county's arginine, and guanidine hydrochloride is (both 0.1-1, especially 0.25-0.75, millimole) or urea, especially 0.5-4 ( 1-3.5) the most rear class of M.