幽门螺杆菌重组Bb-hpaA-vacA疫苗的构建

目的 构建幽门螺杆菌( Hp) 重组双歧杆菌( Bb) Bb-hpaA-vacA疫苗。方法 通过 PCR 分别扩增hpaA和vacA抗原编码基因, 然后采用gene SOEing 剪接hpaA和vacA, 得到hpaA-vacA融合基因;将该融合基因定向克隆到大肠埃希菌-双歧杆菌穿梭表达载体pGEX-1λT, 构建重组质粒 pGEX-hpaA-vacA，电穿孔法将该质粒导入Bb, 构建幽门螺杆菌重组hpaA-vacA 疫苗。结果 PCR成功扩增出分子量约为1500 bp的 hpaA-vacA融合基因，双酶切证实hpaA-vacA融合基因成功插入pGEX-1λT中，并成功转化入双歧杆菌，构建rBb-hpaA-vacA疫苗。结论 成功构建螺杆菌rBb-hpaA-vacA疫苗，为该疫苗的进一步研究奠定了基础。

Construction of the recombinant Bb-hpaA-vacA vaccine of Helicobacter pylori

To construct the recombinant Bb-hpaA-vacA vaccine of Helicobacter pylori（Hp）. hpaA and vacA antigen gene were amplified by PCR and the fusion gene hpaA-vacA was obtained with gene SOEing. Then the fusion gene was cloned into Escherichia coli-Bifidobacteria shuttle plasmid pGEX-1λT to construct pGEX-hpaA-vacA. The recombinant plasmid was electroporated into Bifidobacteria bifidum(Bb) to construct rBb hpaA-vacA vaccine. A 1500bp fusion gene of hpaA-vacA was successfully amplified by PCR and cloned into pGEX-1λT by restriction analysis, and the rBb hpaA-vacA vaccine was successfully constructed by PCR and restriction analysis. In this way, the rBb hpaA-vacA vaccine of Helicobacter pylori is successfully constructed, which lays the experimental foundation of exploitation and utilization of this vaccine.