凋亡素原核表达及纯化

Expression and Purification of Apoptin Gene in E. coil

为进一步研究凋亡素结构与功能和制备凋亡素单克隆抗体,本试验构建了凋亡素的原核表达载体pET-28a-VP3,并将该质粒转化到大肠杆菌E.coliBL21中,以IPTG对其进行诱导表达,聚丙烯酰胺凝胶电泳分析蛋白。结果证明在E.coliBL21中正确表达了凋亡素,同时对表达蛋白进行纯化,SDS-PAGE显示在1.4 kD处出现目的蛋白带与预期相符。

A novel protein named apoptin induces aapoptosis in various human tumor cells but not in normal cells. To study construction and function of apoptin, recombinant expression plasmid pET-28a-VP3 was constructed. Then the recombinants were transfered into the host strain BL21 （DE.3） to induce apoptin fusion protein expression by IPTG when ODroo of the culture was about 0.6. The specific protein expressed was detected by SDS-PAGE. The apoptin expression system with pET-28a-VP3 can effectively express apoptin fusion protein.