用一种新型的PCR方法快速制备人TNF-α缺失突变体  

Applying a New PCR Method to Quickly Prepare Human TNF-α Deletion Mutants

作　　者：张巍[1] 沈倍奋[1] 黎燕[1] 郭宁[1] 冯健男[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《中国生物化学与分子生物学报》2002年第5期600-604,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家杰出青年基金资助课题 (No .3992 5 0 19)~~

摘　　要：人肿瘤坏死因子 α(hTNF α)的 2个Loop区 2 9～ 36、141～ 146是受体的主要结合部位 .应用一种新型的PCR方法 ,快速制备了这 2个Loop区的缺失突变体 ,并对其活性进行了研究 .结果表明 :这种新型的PCR方法在制备缺失突变体中具有快速、简便、经济等优点 ,值得推广 ;缺失蛋白对L92 9细胞的毒性作用明显降低 ,表明这 2个区域为hTNFTumour necrosis factor-α(TNF-α) is a multifunctional cytokine which exerts many effects by binding to cell membrane receptors. It is documented that two loops of human tumour necrosis factor-α(hTNF-α)from 29 to 36 and from 141 to 146,respectively, are major receptor binding sites. A new PCR method was applied to quickly obtain two mutants deleted corresponding loops, respectively, and then the mutants were expressed in E. coli. The cytotoxic activities of the product were studied subsequently. The results indicate that the new method is fast,simple, cheap and deserves popularization, and the cytotoxic activity of the mutants against L929 cells falls down obviously. This demonstrates that the two loops are necessary for hTNF-α to exert its biological activities.

关 键 词：PCR方法 快速制备 人TNF-α缺失突变体 

分 类 号：R394.8[医药卫生—医学遗传学]