幽门螺旋杆菌ureB、vacA基因的原核表达产物及其与感染病人血清的免疫反应性  

Recombinant expression in E.coli and immunoreactivity of ureB and vacA of Helicobacter pylori

作　　者：包士中[1] 史晶[1] 蔡昆[1] 侯晓军[1] 荫俊[1] 王慧[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《胃肠病学和肝病学杂志》2008年第7期552-554,557,共4页Chinese Journal of Gastroenterology and Hepatology

摘　　要：目的在大肠杆菌中重组表达幽门螺旋杆菌UreB、VacA蛋白,并对目的产物与感染病人血清的反应性进行验证。方法PCR扩增获得ureB、vacA基因,分子生物学方法将基因片段克隆入pET22b(+)表达载体并在大肠杆菌中诱导表达,表达产物经Ni离子亲和层析进行纯化,纯化产物采用Western Blot方法鉴定其与感染病人血清的免疫反应性。结果构建的重组大肠杆菌高效表达UreB、VacA蛋白,Ni离子亲和层析后获得90%以上纯度的目的蛋白,与感染病人血清具有良好的免疫反应性。结论重组UreB、VacA蛋白可作为幽门螺旋杆菌感染检测的候选分子。Objective To express the UreB and VacA of H. pylori in E. coli, and to identify their immunological binding activity with serum from infected patients. Methods The ureB and vacA genes were obtained by PCR from the H. pylori genome, and cloned to the expression vector pET22b（ ＋ ）. The genetically engineered bacteria pET22b（ ＋ ）- UreB/BL21 and pET22b（ ＋ ）-VacA/BL21 expressed the recombinant proteins after induced by IPTG, then the rUreB and rVacA were purified by affinity chromatography. The immunoreactivity of the purified proteins was identified by Western Blot. Results The genetically engineered bacteria expressed the rUreB and rVacA successfully. The purified proteins showed an active binding to the serum from infected patients. Conclusion The recombinant UreB and VacA can be used as candidate targets to detect the H. pylori infection.

关 键 词：尿素酶B亚单位 空泡毒素 重组表达 免疫反应性 幽门螺杆菌 

分 类 号：R378[医药卫生—病原生物学]