肠毒素性大肠杆菌K_(99)-ST_1融合基因的克隆及原核表达  

Cloning and prokaryotic expression of the fusion gene expressing antigen K_(99) and ST_1 of enterotoxigenic Escherichia coli

作　　者：李书光[1] 管宇[1] 肖跃强[1] 王金良[1] 沈志强[1] 

机构地区：[1]山东省滨州畜牧兽医研究院山东绿都生物科技有限公司,山东滨州256600

出　　处：《中国兽医科学》2008年第2期111-115,共5页Chinese Veterinary Science

基　　金：山东省科技攻关计划项目(031020101)

摘　　要：为有效预防肠毒素性大肠杆菌(ETEC)引起的犊牛和羔羊腹泻,将PCR扩增获得的ETEC K99菌毛抗原基因和人工合成的ST1基因片段,借助pUCm-T载体,构建了重组质粒pUCm-K99-ST1,从而获得了融合基因K99-ST1;使用EcoRⅠ+SalⅠ双酶切将该融合基因定向插入表达载体pET-30a中,成功构建了表达载体pET-K99-ST1,将其转入表达菌株BL21(DE3)中,经IPTG诱导,获得31 ku的蛋白;Western-blot-ting结果显示,该融合蛋白可与K99阳性血清发生特异性反应。To prevent calf and lamb diarrhea caused by enteotoxigenic Escherichia coli （ETEC）, the K99 gene amplified by PCR and the synthesized nucleic acid fragment of ST1 were cocloned into pUCm-T vector to construct recombinant plasmid pUCm-K99-ST1 ,and then the fusion gene K99-ST1 was obtained. The fusion gene K99- ST1 was inserted into expression vector pET-30a to construct recombinant expression vector pET-K99-ST1. The pET-K99- ST1 was transformed into expression strain BL21（DE3） and induced with IPTG. A 31 ku fusion protein was expressed from BL21 （DE3, pET-K99-ST1 ）. The fusion protein specifically reacted with K99 positive sera in Western-blotting test.

关 键 词：大肠杆菌 K99-ST1融合基因 融合蛋白 

分 类 号：S852.612[农业科学—基础兽医学] Q786[农业科学—兽医学]