表达幽门螺杆菌尿素酶B亚单位的减毒沙门氏菌工程菌株的构建

Construction of an attenuated Salmonella typhimurium vaccine strain expressing urease B subunit of helicobacter pylori

目的采用平衡致死系统构建表达幽门螺杆菌尿素酶 B亚单位的减毒沙门氏菌工程菌株。方法 PCR扩增幽门螺杆菌尿素酶 B亚单位 ,与 p YA 3149(asd+ )载体质粒重组 ,转化减毒的伤寒沙门氏菌突变株 ,SDS- PAGE,用 western- blot分析其表达 ,并观察重组菌体外传代培养的稳定性。结果利用宿主 -载体平衡致死系统构建了表达幽门螺杆菌尿素酶 B亚单位的重组减毒沙门氏菌工程菌株 ,且在有选择压力的条件下在体外能稳定地繁殖、生长和传代。

ObjectiveTo construct an attenuated salmonella typhimurium vaccine strain expressing urease B subunit of H.pylori with balanced lethal system MethodsThe gene of urease B subunit of H pylori was amplified and recombined with a vector(asd +) The recombinant was transformated into attenuated salmonella typhimurium The expression of urease B subunit was analysed by SDS PAGE and western blot, also the stability of vaccine strains was investigated by generation culture in vitro ResultsAn attenuated salmonella typhimurium vaccine strain expressing urease B subunit of H.pylori with balanced lethal system was constructed, and it was very stable ConclusionsThe attenuated Salmonella typhimurium vaccine strain could probably serve as an oral live vaccine to prevent H pylori infection