3种新型人肿瘤坏死因子衍生物在大肠杆菌中的表达与活性检测  

Expression and activity assay of three kinds of novel human tumor necrosis factor-α derivatives

作　　者：徐春晓[1] 倪道明[1] 张振龙[1] 王春荣[1] 赵秀芬 

机构地区：[1]卫生部北京生物制品研究所

出　　处：《中华微生物学和免疫学杂志》2000年第2期132-135,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的　研制高抗瘤活性的新型肿瘤坏死因子。方法　在总结前人有关hTNF α结构与功能关系研究的基础上 ,应用PCR技术构建了 3种多位点同时突变的hTNF α新型编码基因 ,分别在大肠杆菌中进行表达 ,并用DNA序列分析和Westernblot进行鉴定 ,L92 9细胞检测体外抗瘤活性。结果　 3种衍生物均在大肠杆菌中获得高效表达 ,活性测定结果表明 ,3种衍生物对L92 9细胞的细胞毒活性分别比原型hTNF α提高 5 76、6 41和 15 3倍。结论　 3种衍生物的抗瘤活性有了较大的提高 。Objective Studies of novel human tumor necrosis factor α(hTNF α) derivatives with increased antitumor activity. Methods Based on the pervious studies of hTNF α structure functional relationship, three mutated hTNF α genes were generated by PCR. These genes were inserted into E.coli and confirmed by DNA sequencing and Western blot. The antitumor activities were determined by mouse L929 cells. Results Three genes were highly expressed in E.coli . The in vitro cytotoxicity effects of hTNF α derivatives on L929 cells were 576, 641, 153 times higher than that of natural hTNF α. Conclusion The antitumor activities of hTNF α derivatives are highly improved. And the derivatives have displayed potential prospect for clinical application.

关 键 词：肿瘤坏死因子 聚合酶链反应 基因表达 大肠杆菌 

分 类 号：R392.11[医药卫生—免疫学]