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类志贺毒素Ⅱ型变异体A亚单位基因缺失株的构建及表达

徐建生， 陈雷， 成大荣， 董国雄， 李俊宝

（扬州大学 兽医学院， 江苏 扬州225009）

摘要： 采用PCR方法分别对类志贺氏菌毒素A亚单位基因第298～805序列、818～1201序列进行了扩增，使其第806～817序列的碱基（编码167～170位氨基酸）缺失，并将两片段连接后克隆至质粒载体pGEX6P1中，获得了含有重组质粒pGEXAde的重组大肠埃希氏菌；经SDSPAGE以及使用特异性抗SLTⅡeA单克隆抗体的Westernblotting，证明该重组大肠埃希氏菌在IPTG诱导条件下可表达SLTⅡeA。

关键词：水肿病； 志贺样毒素Ⅱ型变异体； A亚单位； 基因缺失

中图分类号：S 852.613：Q786  文献标识码： A  文章编号：1000-6419(2004)12-0018-04

Construction and expression of A subunit gene of Shigalike toxin typeⅡ variant

XU Jiansheng, CHEN Lei, CHENG Darong, DONG Guoxiong, LI Junbao

(College of Veterinary Medicine, Yangzhou University, Yangzhou 225009,China)

Abstract:The 298-805bp and the 818-1201bp fragments of A subunit gene of Shigalike toxin typeⅡ variant (SLTⅡe A gene) were amplified by PCR, and the 806-817bp fragment of the latter fragment was deleted by PCR. The PCR products were cloned into prokaryotic expression vector pGEX6P1 and the recombinant plasmid pGEXAde was constructed. The recombinant plasmid was transformed into E.coli BL21. SDSPAGE and Westernblotting tests confirmed that the fusion protein named GSTⅡeA had been expressed in E.coli BL21 from the SLTⅡeA gene.

Key words: edema disease; SLTⅡe; subunit A; gene deletion

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