中国免疫学杂志Issue(3)：358-363,6.DOI:10.3969/j.issn.1000-484X.2015.03.015
减毒猪霍乱沙门菌ΔcrpΔcyaΔasdC78-1（pYA3493）宿主-载体平衡致死系统的构建及其生物学特性研究
Construction and characterization of host-vector balanced lethal system of attenuated salmonella choleraesuisΔcrpΔcyaΔasdC78-1 (pYA3493)
摘要
Abstract
Objective:In order to develop an oral live vaccine vector of swine that can stably carry exogenous genes.Methods:Mutant ΔcrpΔcyaΔasdC78-1 was constructed by the method of suicide plasmid pREasd-mediated bacteria homologous recombination on the basis of attenuated Salmonella choleraesuisΔcrpΔcyaC78-1.Complementary plasmid pYA3493 with asd was electrotransformed into the mutant,and thenΔcrpΔcyaΔasdC78-1(pYA3493) host-vector balanced lethal system was constructed.Its biological characteristics were analyzed further.Results:The results of PCR and sequencing showed thatΔcrpΔcyaΔasdC78-1(pYA3493) was constructed suc-cessfully.Biological characteristics showed that the serotype of ΔcrpΔcyaΔasdC78-1(pYA3493) was identical to ΔcyaΔasdC78-1 and vaccine strain C500 and it can stably carry theΔasd gene in vitro;its growth speed was a little slower than ΔcrpΔcyaC78-1 strain,but both of their growth speeds were significantly slower than vaccine strain C500;the biochemical characteristics of ΔcrpΔcyaΔasdC78-1 ( pYA3493 ) were basically the same with ΔcrpΔcyaC78-1 strain.Oral virulence test in mice showed that the virulence ofΔcrpΔcyaΔasdC78-1 ( pYA3493 ) was similar with ΔcrpΔcyaC78-1, but its median lethal dose is 412 times of vaccine strain C500.Conclusion:These results demonstrated that attenuated Salmonella choleraesuisΔcrpΔcyaΔasdC78-1(pYA3493) strain had the potential to be used as an oral live vaccine vector for expressing foreign genes efficiently.关键词
自杀性质粒/宿主-载体平衡致死系统/ΔcrpΔcyaΔasdC78-1(PYA3493)/口服活疫苗载体Key words
Suicide plasmid/Host-vector balanced lethal system/ΔcrpΔcyaΔasdC78-1(pYA3493)/Oral live vaccine vector分类
农业科技引用本文复制引用
尚珂,张俊峰,程相朝,张春杰,李银聚,陈桂华,颜云飞,翟崇凯,赵战勤..减毒猪霍乱沙门菌ΔcrpΔcyaΔasdC78-1（pYA3493）宿主-载体平衡致死系统的构建及其生物学特性研究[J].中国免疫学杂志,2015,(3):358-363,6.基金项目
本文为河南省科技攻关项目（112102110019）。 （）
