减毒猪霍乱沙门菌△cya△asdC78-1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 choleraesuis △cya△asdC78-1

作　　者：张俊峰[1] 张春杰[1] 程相朝[1] 尚珂[1] 李银聚[1] 吴庭才[1] 刘一尘[1] 颜云飞 陈桂华[1] 赵战勤[1][1] 

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

出　　处：《中国预防兽医学报》2015年第5期366-370,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：河南省科技攻关项目(112102110019)

摘　　要：为研制能够稳定携带外源基因的猪霍乱沙门菌口服活疫苗载体,本研究在减毒猪霍乱沙门菌△cyaC78-1基础上,利用自杀性质粒介导的等位交换技术,构建了△cya△asdC78-1宿主-载体平衡致死系统,并对其生物学特性进行了鉴定。PCR及测序结果表明△cya△asdC78-1构建正确,其生物学特性检测结果显示△cya△asdC78-1生化特性与△cyaC78-1完全相同,血清型与△cyaC78-1和疫苗株C500相同,并且具有稳定的遗传特性;其生长速度与△cya C78-1基本相同,均明显慢于疫苗株C500。动物试验表明△cya△asdC78-1电转化携带asd基因的互补质粒pYA3493后,其毒力比△crp△asdC78-1(p YA3493)降低1.14倍,比△asdC500(pYA3493)降低了5.1倍。以上结果表明,减毒猪霍乱沙门菌△cya△asdC78-1同样可以作为口服活疫苗载体用于高效表达外源基因,为开发以C78-1为基础的口服多价疫苗载体奠定了基础。Abstract： To develop an oral live vaccine vector of Salmonella choleraesuis for swine which was able to stably carry exogenous genes, the S.choleraesuis △ cya △ asdC78-1 host-vector balanced lethal system was constructed with the method of recombinant suicide plasmid-mediated allelic exchange on the basis of attenuated S.choleraesuis △ cyaC78-1, and the biological characteristics of △ cya △asdC78-1 showed that the biochemical characteristics and the serotype of A cya A asdC78-1 were exactly the same with △cyaC78-1 strain; The growth rate of A cyaA asdC78-1 was basically the same as △cyaC78-1 strain, but both of their growth speeds were significantly slower than vaccine strain C500. Animal experiment showed that the virulence of △ cya ZkasdC78-1 carried complementary plasmid pYA3493 by electroporation was decreased by 1.14 and 5.1 times compared with △ crp △asd C78-1 （pYA3493） and △ asdC500 （pYA3493） in mice. In conclusion, these results demonstrated that attenuated S. choleraesuis △ cya △ asdC78-1 could be used as an oral live vaccine vector for expressing foreign genes efficiently and facilitate for developing an oral polyvalent vaccine vector based on the C78-1 strain.

关 键 词：自杀性质粒 宿主-载体平衡致死系统 △cya△asdC78-1(pYA3493) 口服活疫苗载体 

分 类 号：S852.61[农业科学—基础兽医学]