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.
出处
《胃肠病学和肝病学杂志》
CAS
2008年第7期552-554,557,共4页
Chinese Journal of Gastroenterology and Hepatology
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
Urease B Subunit
Vacuolating cytotoxin
Recombinant expression
Immunoreactivity
Helicobacter pylori
