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目的 构建表达幽门螺杆菌 (Hp)尿素酶A亚单位 (UreA)和过氧化氢酶 (KatA)的减毒鼠伤寒沙门氏菌疫苗株 ,对研制抗Hp感染重组疫苗的可行性进行探讨。方法 PCR方法从Hp基因组中扩增出ureA和katA片段 ,将其插入pGSTag表达载体中 ,重组质粒再转入减毒鼠伤寒沙门氏菌。结果 对重组质粒进行限制酶切分析和PCR检测 ,证实两种基因已被克隆入pGSTag ,并转入减毒鼠伤寒沙门氏菌。结论 本研究成功地将表达HpureA和katA融合基因的重组质粒转入减毒鼠伤寒沙门氏菌中 ,构建了UreA/KatA双价口服活疫苗 ,为进一步研究其在预防Hp感染中的作用奠定了基础
Objective To construct an attenuated Salmonella typhimurium vaccine strain expressing Helicobacter pylori (Hp) A subunit (UreA) and catalase (KatA) and discuss the feasibility of developing a recombinant vaccine against Hp infection. Methods The ureA and katA fragments were amplified from Hp genome by PCR and inserted into pGSTag expression vector. The recombinant plasmids were transformed into attenuated Salmonella typhimurium. Results Restriction analysis and PCR analysis of recombinant plasmids confirmed that both genes had been cloned into pGSTag and transferred into attenuated Salmonella typhimurium. Conclusions This study successfully transferred the recombinant plasmid expressing HpureA and katA fusion gene into attenuated Salmonella typhimurium, and constructed the bivalent UreA / KatA oral live vaccine, which laid the foundation for further study on its role in preventing Hp infection