幽门螺杆菌Ure B串联融合表位的表面呈现表达

Surface Display Expression of Helicobacter pylori Ure B Fusion Epitope

目的:应用表达载体pHIE3N表面呈现表达幽门螺杆菌Ure B串联融合表位。方法:通过引入多克隆位点的方法改造表达呈现表达载体pHIE11,并用改造后的质粒表达幽门螺杆菌Ure B串联融合表位,用SDS-PAGE、Western blot和全菌ELISA检测所得到的重组菌。结果:改造的质粒插入序列正确,能够特异性呈现表达Ure B串联融合表位,表达产物的分子量约为60kDa,与预期大小一致。全菌ELISA结果表明Ure B串联融合表位表达于菌体表面。结论:pHIE3N载体能够表面展示呈现Ure B串联融合表位,构建的重组菌为幽门螺杆菌候选疫苗的研发奠定了基础。

Objective：To achieve surface display of H.pylori ure B fusion epitope by vector pHIE3N.Methods：Multiple cloning site was introduced into pHIE3N in order to express the H.pylori ure B fusion epitope protein.The recombinant strain was confirmed by SDS-PAGE,Western blot and ELISA.Result：The inserted sequence of the reconstructed plasmid was correct.The molecular weight of the fused Ure B protein was about 60 kDa which had proved by SDS-PAGE and Western blot analysis,ELISA results showed that the Ure B fusion epitope expressed on cell surfaces.Conclusion：The Ure B fusion epitope was successfully expressed on the sruface of E.coli DH5α using pHIE3N surface display vector.This result will contribute to the vaccine research of H.pylori.