假结核耶尔森菌侵袭素蛋白基因的克隆及原核表达Gene Cloning and Prokaryotic Expression of Invasin Protein of Yersinia pseudotuberculosis
杨春静,芦清霞,克力比努尔·热合曼,刘红春,夏雪琴,李江伟
摘要(Abstract)：
目的克隆假结核耶尔森菌侵袭素蛋白基因并在原核表达系统中表达.方法根据GenBank上登录的假结核耶尔森菌侵袭素蛋白核酸序列设计引物,用PCR方法扩增假结核耶尔森菌侵袭素蛋白基因,并将其插入克隆载体pMD18-T中,测序鉴定正确后再将其克隆到原核表达载体pGEX-4T-1上,得到重组质粒pGEX-4T-1-inv.将重组质粒转化大肠杆菌BL21(DE3)感受态细胞,经IPTG诱导,SDS-PAGE电泳检测.结果假结核耶尔森菌提取的基因组,经PCR扩增,得到一段591 bp的片段,与预期结果一致;SDS-PAGE电泳分析,所得重组蛋白的分子量约46 kDa,与预期结果相同;重组菌体超声裂解后,经12%的SDS-PAGE电泳分析,结果显示该重组蛋白为包涵体蛋白.结论克隆的侵袭素基因核心片段可以在原核细胞中高效表达.
关键词(KeyWords)： 假结核耶尔森菌;侵袭素蛋白;克隆;原核表达
基金项目(Foundation): 新疆维吾尔自治区高校科研计划重点项目(XJEDU2005105)资助
作者(Author): 杨春静,芦清霞,克力比努尔·热合曼,刘红春,夏雪琴,李江伟
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