产肠毒素大肠杆菌F5菌毛基因的克隆与表达

Clone and expression of pili subunit gene from adhesion F5 of the enterotoxigenic Escherichia coli(ETEC)

应用PCR方法从产肠毒素性大肠杆菌(ETEC)中扩增出不含信号肽序列的F5菌毛抗原基因片段,克隆测序后将其连接到大肠杆菌表达载体pET-30a(+)中,转化工程菌BL21,经IPTG诱导表达。经SDS-PAGE和Western Blot分析,重组蛋白在BL21中得到表达并与F5菌毛单因子血清发生明显反应。

A F5 fimbrial subunit gene without signal peptide sequence was amplified by PCR from enterotoxigenic E. coll. After cloning and sequencing, the fragment was inserted into an E. coli expression vector pET- 30a（＋）, then the recombinant plasmid was transferred into E. coli BL21, and E. coli BL21（＋F5） obtained. Cultured and induced with IPTG, the expression of target protein was determined by SDS-PAGE and Western Blot. It was found that the expressed proteins could obviously react with antiserum against the single factor of F5 pili subunit proteins.