3种人肿瘤坏死因子衍生物的研制  

Genetic-engineered Derivatives of the Human Tumor Necrosis Factor

作　　者：何晓龙[1] 常金丽[1] 蔡武城 赵寿元[1] 路长林[1] 朱鹤年[1] 

机构地区：[1]复旦大学遗传学研究所,第二军医大学神经生物学教研室

出　　处：《生物化学与生物物理学报》1995年第1期67-73,共7页

基　　金："863"高技术计划资助;国家自然科学基金

摘　　要：本文在详细分析肿瘤坏死因子（ＴＮＦ）结构的基础上应用ＰＣＲ技术和基因工程手段改造人ＴＮＦ分子，构建了３种人ＴＮＦ的衍生物。３种人ＴＮＦ衍生物是；Ｎ端缺失７个氨基酸且８、９、１０位的ＰｒｏＳｅｒＡｓｐ改为ＡｒｇＬｙｓＡｒｇ的ＴＮＦ（简写为ｈＴＮＦＤ１）：Ｃ端１５７位Ｌｅｕ改为Ｐｈｅ的ＴＮＦ（简写为ｈＴＮＦＤ２）；Ｎ端和Ｃ端同时作上述改变的ＴＮＦ（简写为ｈＴＮＦＤ３）。这３种衍生物在大肠杆菌中均获得较高表述，它们对Ｌ９２９细胞的细胞毒活性较重组天然人ＴＮＦ有很大升高，尤其是ｈＴＮＦＤ１和ｈＴＮＦＤ３，升高达３个数量级。文中对３种衍生物活性升高的原因作了分析。Based on the detailed analysis of TNF structure,we have developed three human TNF derivatives with PCR and genetic engineering techniques.The three hTNF derivatives are:hTNFD1 in whieh seven N-terminal amino acids of hTNF are deleted and ProSerAsp at positions 8, 9, 10 are substituted by ArgLysArg;hTNFD2 in which Len at position 157 of hTNF is substituted by Phe; hTNFD3 which contains both alterations mantioned above.The three TNF derivatives are highly expressed in E. coli. Their cytotoxic activity to L929 cell line is much higher than that of hTNF, especially hTNFD1 and hTNFD3, whose eytotoxie activity has inereased 3 orders of magnitude as compared to that of hTNF.In respect of the enhancement of eytotoxic aetivity hTNFD1 and hTNFD3 greatly exceed any other TNF derivatives previously reported. The mechanism of increase in cytotoxie aetivity of TNF derivatives is discussed.

关 键 词：肿瘤坏死因子 衍生物 制备 聚合酶链反应 

分 类 号：R392－33[医药卫生—免疫学]