大肠杆菌K88菌毛蛋白faeG亚基基因克隆及表达  

Cloning and expression of Escherichia coli K88 fimbrial subunit faeG gene

作　　者：徐建生[1] 成大荣[1] 任士飞[1] 孙怀昌[1] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2005年第3期1-4,共4页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家高技术研究发展计划项目(8632003AA222141)

摘　　要：利用PCR技术,从致仔猪黄痢的大肠杆菌中扩增不含信号肽序列的K 88菌毛蛋白faeG亚基基因片段,将其克隆到表达载体pGEX-6P-1中,构建该基因的原核表达载体pGEX-F aeG,通过测序证明序列正确后,导入大肠杆菌BL 21,得到工程菌株PGEX-F aeG。IPTG诱导表达,SDS-PAGE分析结果表明,融合蛋白(约53 ku)在大肠杆菌BL 21中得到高效表达,表达产物约占菌体蛋白的35%,免疫印记结果表明此融合蛋白与K 88单抗反应。The DNA fragment of K88 fimbrial subunit faeG gene without signal peptide sequence was amplified by PCR from the DNA of enterotoxigenic Escherichia coli K88 strain. Seqnence cloned faeG gene was 795 bp and share 95% identity with the pulished sequence. The PCR product was digested by restriction enzymes and then subcloned into the prokaryotic expression vector pGEX-6p-1. The GST-FaeG fusion protein of about 53 ku was expressed in E. coli BL21 by IPTG induction and was revealed by SDS-PAGE analysis, which was confirmed by wester blotting.

关 键 词：大肠杆菌 K88菌毛 克隆 表达 

分 类 号：Q785[生物学—分子生物学] S852.61[农业科学—基础兽医学]