表达幽门螺杆菌UreA和KatA双价抗原的鼠伤寒菌疫苗株的初步构建

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

CONSTRUCTION OF SALMONELLA TYPHIMURIUM BIVALENT VACCINE STRAIN EXPRESSING UreA AND KatA

Aim To construct the recombinant attenuated Salmonella typhimurium vaccine strain expressing urease A subunit(UreA) and catalase(KatA) of Helicobacter pylori (Hp), and investigate the feasibility of developing a recombinant vaccine against Hp infection.Methods ureA and katA were amplified by PCR from Hp genome and cloned into expression vector pGSTag.This recombinant plasmid was transformed into attenuated Salmonella typhimurium. Results With restriction enzyme digestion and PCR,it was showed that ureA and katA have been inserted into pGSTag and introduced into the attenuated Salmonella typhimurium.Conclusion We were successful in transforming the recombinant plasmid expressing the ureA and katA fusion genes into attenuated Salmonella typhimurium and constructing a ureA / katA bivalent live oral vaccine strain . Our study laid foundation of further studying the effect of this vaccine on prevention and treatment of Hp infection.