幽门螺杆菌CrdR蛋白的原核表达  

Prokaryotic expression of CrdR protein of Helicobacter pylori

作　　者：唐磊[1] 杨致邦[2] 聂佳莹[1] 黄进[1] 

机构地区：[1]重庆医科大学神经科学研究中心,重庆400016 [2]重庆医科大学基础医学实验教学中心病原生物学与免疫学实验室,重庆400016

出　　处：《中国生物制品学杂志》2013年第9期1260-1263,共4页Chinese Journal of Biologicals

摘　　要：目的原核表达幽门螺杆菌(Helicobacter pylori,H.pylori)CrdR蛋白(即HP1365),以探讨其在酸适应中的调控机制。方法从H.pylori 26695标准株基因组DNA中PCR扩增hp1365基因,克隆至表达载体pGEX-6P-1中,转化E.coli JM109,IPTG诱导表达,表达产物经SDS-PAGE及Western blot鉴定。结果重组表达质粒pGEX-hp1365经双酶切、PCR和测序鉴定构建正确;表达的重组蛋白相对分子质量约为25 000,主要以包涵体形式表达,可与鼠抗His标签单克隆抗体特异性结合。结论原核表达了H.pylori CrdR蛋白,为进一步探讨其对酸信号的感应机制及其通过阻断酸信号的感应抗H.pylori感染的途径奠定物质基础。Objective To express CrdR protein （HP1365） in prokaryotic cells and investigate the mechanism of its regu- latory effect in acidic adaptation. Methods The hp1365 gene was amplified from genomic DNA of Helicobacter pylori standard strain 26695 by PCR and cloned into expression vector pGEX-6P-1. The constructed recombinant plasmid was transformed to E. coli JM109 and induced with IPTG. The expressed product was identified by SDS-PAGE and Western blot. Results Restriction analysis, PeR and sequencing proved that recombinant plasmid pGEX-hp1365 was constructed correctly. The expressed recombinant protein, with a relative molecular mass of about 25 000, mainly existed in a form of inclusion body and showed specific binding to mouse monoclonal antibody against His tag. Conclusion The CrdR pro- tein of H. pylori was successfully expressed, which laid a foundation of further study on mechanism of inductivity of CrdR to acid signal as well as the measures of anti-H, pylori infection by blocking the inductivity.

关 键 词：幽门螺杆菌 酸适应 CrdR蛋白 原核细胞 基因表达 

分 类 号：R378.99[医药卫生—病原生物学] Q786[医药卫生—基础医学]