表达恶性疟原虫裂殖子表面蛋白-1片段的减毒鼠伤寒杆菌的构建

目的 检测恶性疟原虫裂殖子表面蛋白１基因在减毒鼠伤寒直蓖Ｘ４０６４株中的表达。方法 将７个克隆有恶性疟原虫裂殖子表面蛋白１基因片段的表达质粒ｐＱＥＭ用氯化钙法转化到修饰系统正常、限制系统缺陷的鼠伤寒杆菌ＬＢ５０００中，再通过专一性噬菌体Ｐ２２转导入腺苷酸环化酶基因、环腺苷酸受体基因双重突变的减毒鼠伤寒杆菌Ｘ４０６４株中，用质粒酶切、Ｗｅｓｔｅｒｎ ｂｌｏｔ鉴定构建的重组菌株 组菌株的表达。结果 成

Construction of recombinant attenuated Salmonella typhimurium vaccine strain expressing Plasmodium falciparum merozoite surface protein-1 fragments

Objective To examine the expression of merozoite surface protein-1(MSP-1)gene of Plasmodium falciparum merozoite in attenuated Salmonella typhimurium X4064 strain.Methods Sevan pQEM plasmids cloned with MSP-1 gene fragments were firstly transformed into S,typhimurium LB5000,a restriction-negative,modification.positive strain,to be modified and then transduced into△cya △crp double mutant S.typhimurium X4064 by using bacteriophage P22.Restriction enzyme analysis and Western blot were used to examine the construction and expression of these recombinant strains.Results Seven recombinant strains of S.typhimurium X4064(pQEM)were successfully constructed.The MSP-1 protein fragments were expressed in these recombinant strains.Conclusion The recombinant S.typhimurium X4064(pQEM)strains could be considered as candidates for malaria vaccine.