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目的:构建携带人幽门螺杆菌(H pylori)中性粒细胞激活蛋白(HP-NAP)基因(napA)的重组活减毒鼠伤寒沙门菌口服DNA 疫苗. 方法:应用基因工程技术扩增全长napA,测序并同源性分析后,将其亚克隆入真核表达载体pIRES,鉴定正确后将重组质粒转化活减毒鼠伤寒沙门菌. 结果:重组质粒经PCR及双酶切,证实成功构建了携带HP-NAP基因的重组真核表达质粒pIRES-napA,后者成功转化活减毒鼠伤寒沙门菌SL7207.所克隆435bp napA与GenBank中SS1-napA核苷酸和蛋白质的同源性均为98%. 结论:成功构建并鉴定了携带HP-NA2P基因的重组活减毒鼠伤寒沙门菌口服DNA疫苗,为多价抗H pylori口服DNA疫苗的研制奠定了基础.
Objective: To construct recombinant live attenuated Salmonella typhimurium oral DNA vaccine carrying human Helicobacter pylori (HP-NAP) gene (napA) .Methods: The full length napA was subcloned into the eukaryotic expression vector pIRES after sequencing and homology analysis.The recombinant plasmids were transformed into live-attenuated Salmonella typhimurium.Results: The recombinant plasmids were successfully constructed by PCR and double enzyme digestion The recombinant eukaryotic expression plasmid pIRES-napA carrying HP-NAP gene was successfully transformed into live-attenuated Salmonella typhimurium SL7207.The homology between the cloned 435 bp napA and SS1-napA nucleotide and protein in GenBank was 98% .Conclusion: The oral DNA vaccine of recombinant live-attenuated Salmonella typhimurium carrying HP-NA2P gene was successfully constructed and identified, which laid the foundation for the development of multivalent anti-H pylori oral DNA vaccine.