幽门螺杆菌CrdS蛋白的原核表达和初步鉴定  

Prokaryotic Expression and Preliminary Identification of Protein CrdS of Helicobacter Pylori

作　　者：聂佳莹[1] 杨致邦[2] 唐磊[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：《生物医学工程学杂志》2013年第6期1298-1301,共4页Journal of Biomedical Engineering

摘　　要：原核表达幽门螺杆菌(Helicobacter pylori,H.pylori)酸适应感应蛋白CrdS,探讨其酸适应的调控机制。提取H.pylori26695标准株全基因组DNA作为模版,PCR扩增编码CrdS蛋白的基因hp1364,构建重组克隆质粒pUCm-T-hp1364和原核表达质粒pQE30-hp1364,经PCR、双酶切和测序鉴定正确后,转化大肠埃希菌XL1blue,IPTG诱导表达CrdS蛋白。结果表明:通过Western blot鉴定和SDS-PAGE分析该重组蛋白的相对分子质量约为46kDa,主要以包涵体形式存在。原核表达并成功获得的H.pylori CrdS蛋白,能够为探讨其酸适应的调控机制和新的抗H.pylori感染的途径提供实验材料。The IZrdb protein responding to the acidic adaptation was prokaryotic-expressed in our Laboratory to ex- plore the regulatory mechanism in the acidic adaptation of helicobacter p2alori （H. pylori）. The whole genomic DNA of H. pylori strain 26695 was abstracted and set as the template firstly. And then the hp1364 gene coding CrdS pro- tein was amplified via the PCR technique. Then the clonal recombinant plasmid pUCm T hp1364 and the prokaryotic expression plasmid pQg30 hp1364 were built and identified by the methods of PCR, cutting with two enzymes and sequencing. After that, the plasmid pQE30-hp1364 was transferred into the E. coli XLlblue and induced with IPTG. Using western blot and SDS-PAGE, it can be analyzed that the expressed recombinant protein existed mainly in the form of the inclusion bodies and its relative molecular mass was about 46 kDa. The successfully attained recombinant protein CrdS will provide the material to explore the regulatory mechanism in the acidic adaptation of H. pylori and the new way to resist the infection of H. pylori.

关 键 词：幽门螺杆菌 酸适应 感应蛋白 原核表达 

分 类 号：R378[医药卫生—病原生物学]