摘要
[目的]研究重组麻疯树核糖体失活蛋白的表达及其体外抗肿瘤活性。[方法]从麻疯树叶中克隆了curcin基因,在此基础上构建pET-22b/curcin重组表达载体,在大肠杆菌中成功表达29 KD的重组蛋白。[结果]经0.5 mM的IPTG在30℃诱导6 h,重组蛋白表达量约占菌体总蛋白的20%,重组curcin以包涵体的形式表达。经溶解、复性、纯化,获得了纯度为95.6%的重组curcin蛋白。[结论]体外研究表明,重组蛋白对A549、HepG2的肿瘤细胞有明显的增殖抑制作用,其IC50分别为92.246和691.42μg/ml,但是对HELA细胞增殖没有明显抑制作用。
[Objective] To study the expression of recombinant curcin in Jatropha curcas L.and its antitumour activity in vitro.[Method] Curcin was cloned from the leaves of J.curcas,and the recombinant vector pET-22b was established.The recombinant protein（29KD） was expressed in E.coli.[Result] Induced by 0.5 mM IPTG for 6 h at 30 ℃,expressing quantity of recombinant protein was about 20% of the total protein.Recombinant curcin was expressed in the form of inclusion body.After dissolution,renaturation and purification,purity of recombinant curcin reached 95.6%.[Conclusion] In vitro research showed that recombinant protein had significant proliferation inhibition effects on the A549 and HepG2 tumor cells.And their IC50 were 92.246 and 691.42 μg/ml,respectively.But the recombinant protein had no significant inhibitory effects on Hela.
出处
《安徽农业科学》
CAS
2012年第12期7079-7082,共4页
Journal of Anhui Agricultural Sciences
