【摘要】 化学合成的人尿激酶原ｃＤＮＡ克隆在表达质粒ｐＥＴ１１ｄ中，在Ｔ７启动子的作用下，经０１ｍｍｏｌ／ＬＩＰＴＧ诱导，在大肠杆菌ＢＬ２１（ＤＥ３）ｐＬｙｓＳ中获得表达。其表达量占菌体总蛋白的１５％，表达产物以无活性的包涵体形式存在。经体外变复性，Ｚｎ２＋选择性沉淀，抗体亲和柱层析，及Ｂｅｎｚａｍｉｄｉｎｅ亲和吸附，所表达的人尿激酶原被纯化为单一条带，其比活约１１００００ＩＵ／ｍｇ。更多还原

【Abstract】 A chemically synthesized human pro urokinase (pro UK) cDNA was cloned into the expression vector pET 11d,and expressed in E.coli BL21(DE3) pLysS under the control of T7 promoter.By using 0.1mmol/L IPTG induction,the expression level of the recombinant pro UK was over 15% of total bacterial proteins as inclusion bodies.After denaturation and renaturation in vitro ,the expressed pro UK was purified to Identity by Zn 2+ selective precipitation,immuno affinity chromato graphy,and Benzamidine affinity adsorption.The specific activity of the purified human pro UK was about 110000IU/mg.更多还原