人幽门螺杆菌UreB与Omp11双价疫苗载体的构建与鉴定

Construction of recombinant vector of bivalent vaccine with fusion gene ureB-omp11 of human Helicobacter pylori

目的:构建含人幽门螺杆菌(H.pylori,Hp)郑州分离株MELHP27ureB-omp11融合基因的重组表达载体。方法:利用分子克隆技术,扩增HpUreB、Omp11编码基因及融合基因ureB-omp11,将融合基因ureB-omp11与载体pET30a(+)进行酶切、连接,然后转化并筛选含有目的基因的重组载体。结果:酶切、测序分析表明,插入的基因片段为HpureB-omp11融合基因,由2280bp组成。与GenBank报道的相比,核苷酸序列同源性为99.39%,氨基酸序列同源性为99.47%。结论:成功构建了MELHp27融合蛋白UreB-Omp11的双价疫苗重组载体,为Hp蛋白质疫苗和核酸疫苗的研制奠定了良好的基础。

Aim ： To construct a recombinant vector with ureB-omp1 1 fusion genc of human H, pylori MEL Hp27 isolated from a certain patient in Zhengzhou. Methods：The target genes encoding UreB and Omp1 1 were amplified by PCR. The fusion gene ureB-omp1 1 was amplified by overlap extension PCR and digested by restricted endonuclease enzyme and inserted into the corresponding endonuclease enzyme digested prokaryotic expression vector pET30a（ ＋ ）. The recombinant vector pET3Oa（ ＋ ）-ureB-omp1 1 was identified by PCR and restricted endonuclease enzyme and sequence analysis. Results： Enzyme digestion analysis and sequencing showed that the target genes were 2 280 bp and had been inserted into recombinant vector. Compared with gene reported by GenBank, the target genes had 99.39% homology in nucleotide acid sequence and 99.47% homology in amino acid sequence. Conclusion： The recombinant expression vector with ureB-omp1 1 fusion gene of H. pylori MEL Hp27 has been constructed successfully, which lays a good foundation for development of Hp protein and DNA vaccine applying to prevent Hp infection.