噬菌体显示法克隆RNA结合蛋白cDNA——表达文库的构建

Cloning cDNA of RNA-binding Proteins by Using Phage Display Method:Construction of the Phage Expression Library

试图用噬菌体显示法 (phagedisplay)克隆编码与人白细胞介素 6核转录因子的 3′非翻译区(NF IL63′UTR)特异结合的蛋白的cDNA .构建了回复系RR细胞的cDNA的噬粒 ( phagemid)表达文库 ,并设法除去了cDNA的大部分终止密码子 .鉴定表明 ,该文库中含有各种大小的cDNA插入片段 ,并能够表达外源cDNA 。

In order to clone cDNAs coding for proteins specially binding to NF-IL6's 3′UTR, a phagemid expression library was constructed from cDNAs of revertant RR cells. The termination codons of those cloned cDNAs were largely removed by restricted exoⅢ enzyme digestion, in order to facilitate the expression of cDNA as fusion proteins with the phage gene Ⅲ. Examinations of this library showed that the library includes various lengths of cDNA inserts and it can express exogenous cDNA.