鼠伤寒沙门菌SL1344株侵袭性蛋白B缺失株asd平衡致死系统的构建及特性的研究  

Construction and characterization of invasion protein B gene deleted mutant asd host-vector balanced lethal system

作　　者：张春杰[1] 陈松彪[1] 程相朝[1] 廖成水[1] 李静[1] 何雷[1] 张明亮[1] 郁川[1] 余祖华[1] 贾艳艳[1] 赵战勤[1] 

机构地区：[1]河南省动物疫病与公共安全院士工作站/河南科技大学动物疫病与公共卫生重点实验室,洛阳471003

出　　处：《中国免疫学杂志》2016年第2期210-213,共4页Chinese Journal of Immunology

基　　金：河南省科技攻关项目(152102110078)资助

摘　　要：目的:为研究用于能够稳定携带外源基因的口服活疫苗载体。方法:本研究以减毒鼠伤寒沙门菌SL1344ΔsipB为研究对象,利用自杀性质粒pREΔasd介导的细菌同源重组技术,构建SL1344ΔsipBΔasd宿主-载体平衡致死系统,并进一步研究其生物学特性。结果:PCR及测序结果表明SL1344ΔsipBΔasd构建成功。生物学特性结果表明SL1344ΔsipBΔasd的血清型与SL1344ΔsipB相同,并能够稳定遗传缺失后的asd基因;其生长速度和生化特性与亲本株SL1344基本相同,并且该缺失株仍然具有运动性的能力,小鼠攻毒试验表明SL1344ΔsipBΔasd电转了携带asd基因的互补质粒pYA3493后,其毒力较亲本菌株SL1344约降低至1.4%。结论:以上结果证明,减毒鼠伤寒沙门菌SL1344ΔsipBΔasd菌株构建成功,为深入研究以鼠伤寒沙门菌为载体的口服多价疫苗奠定基础。Objective： To develop an oral live vaccine vector which stably carries exogenous genes. Methods： SL1344AsipBAasd host-vector balanced lethal system was constructed by the method of recombinant suicide plasmid-mediated allelic exchange on the basis of attenuated Salmonella typhinurium SL1344AsipB. Then, the biological characteristics of SL1344AsipBAasd was analyzed. Results： The results showed that the mutant was stabile with the Aasd gene in vitro; the serotype and growth rate of SL1344AsipBAasd strain was almost same as the parent SL1344AsipB and SL1344 strain. And the mutant strains remain swim ruing zones. Virulence test in mice showed that the virulence of SL1344AsipB△asd which carried complementary plasmid pYA3493 by electrotransformation decreased by 1.4% compared with SL1344. Conclusion： These results showed that the SL1344AsipBAasd mutant was successfully constructed. It is likely that this mutant should be used as a live vector to express foreign genes.

关 键 词：自杀性质粒 宿主-载体平衡致死系统 SL1344ΔsipBΔasd(pYA3493) 口服活疫苗载体 

分 类 号：S852.612[农业科学—基础兽医学]