携带幽门螺杆菌hpaA基因减毒鼠伤寒沙门疫苗菌的构建

构建携带幽门螺杆菌（H.pylori)hpaA基因的重组活减毒鼠伤沙门疫苗菌。方法：用分子生物学方法将hpaA基因克隆入原核表达质粒pTrc99A,并进行核苷酸测序,重组质粒经再导入活减毒鼠伤寒沙门菌SL3261,提取重组菌苗质粒,聚合酶链反应（PCR）和酶切鉴定,筛选目的克隆。结果：经PCR和酶切证实,构建了携带hpaA基因（560bp)的重组核表达质粒pTrc99A-hpaA,并将后者成功转化活减毒鼠伤寒沙门菌SL3261。结论：S我建并鉴定了携带H.pylorihpaA基因的重组活减毒鼠伤寒沙门疫苗菌,为探索制备H.ylori口服份活疫苗奠定了基础。

Construction of an Attenuated Salmonella typhimurium Vaccine Strain Carrying Helicobacter pylori hpaA Gene

Background/Aims: To construct a recombinant live attenuated Salmonella typhimurium vaccine strain carrying Helicobacter pylon (H. pylon) hpaA gene. Methods: By means of molecular biology, hpaA gene was amplified from H. pylon genome DNA and cloned into a prokaryotic expression plasmid pTrc99A. After sequence analysis, the identified recombinant plasmid was then used to transform a live attenuated Salmonella typhimurium vaccine strain SL3261, and the positive clones were screened by polymerase chain reaction (PCR) and restriction enzyme digestion. Results: Confirmed by the above methods, a recombinant procaryotic expression plasmid pTrc99A-hpaA harbouring about 560 bp hpaA gene was constructed, and the recombinant plasmid was then introduced into a live attenuated Salmonella typhimurium vaccine strain SL3261 successfully. Conclusions: A recombinant live attenuated Salmonella typhimurium vaccine strain harbouring H. Pylon hpaA gene was constructed and identified; this work will help developing an oral recombinant live vaccine against H. pylon infection.