3′端非翻译区影响TNFα cDNA在大肠杆菌中的表达  

Effect of 3'-untranslated Region on Expression of TNFa cDNA in Escherichia coli

作　　者：常金丽[1] 蔡武城 徐立峰 李昌本[1] 赵寿元[1] 

机构地区：[1]复旦大学遗传学研究所遗传工程国家重点实验室,上海200433

出　　处：《生物工程学报》1995年第4期304-309,共6页Chinese Journal of Biotechnology

基　　金：863计划基金资助项目

摘　　要：采用PCR方法改变了表达构建物pRL-rhTNF中的结构,即去掉rhTNFαcDNA3＇端非翻译区序列110bp(命名为pRL-rhTNFα2),转入大肠杆菌后,观察数个阳性转化子的表达情况,并与原型pRL-rhTNFα的表达进行比较。结果表明,去掉3＇端非翻译区的pRL-rhTNFα2能稳定表达rhTNFα,其发酵及纯化产品的生物学特征均等同于原型的,且其表达量比原型的有提高,提示3＇端非翻译区序列对表达有影响。3＇端非翻译区内存在一个相似于TA的重复序列——TTTA TTA七聚体。这可能是本研究中影响TNFα表达量的原因之一。A construct pRL-rhTNFa2, in which the 110bp 3'-untranslated region of rhTNFa c cDNA was deleted, was transformed in E. colt. The expression level of several positive transformants was determined. The results showed that the expression of pRL-TNFa2 was stable and the biological characteristics of the protein product were as same as that of pRL-TrhTNFa. Furthermore, the expression level of pRL-rhTNFa2 was increased. It suggested that 3'-untranslated region may have some negative effects on gene expression. A TA-rich sequence, TTTATTA, contained in 3'-untranslated region of pRL-rhTNFa may be involved in the inhibitory effect on gene expression.

关 键 词：肿瘤坏死因子A CDNA 3'端非翻译区 基因工程 

分 类 号：R392－33[医药卫生—免疫学] R394.8[医药卫生—基础医学]