表达PRRSV GP3蛋白减毒鼠伤寒沙门氏菌疫苗株的构建  

Construction of the Attenuated Salmonella typhimurium Vaccine Strains Expressing PRRSV GP3 Antigen

作　　者：赵红妮[1] 许信刚[1] 童德文[1] 罗小毛[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100

出　　处：《西北农业学报》2011年第10期6-11,共6页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：陕西省农业科技创新项目(2010NKC-06);国家大学生创新性实验计划项目(2009);教育部新世纪优秀人才支持计划(NCET-07-0701)

摘　　要：构建表达猪PRRSV GP3蛋白的口服重组减毒鼠伤寒沙门氏菌活载体疫苗株。将去信号肽序列的PRRSV ORF3基因插入到表达载体pYA3341(Asd+)中,构建重组质粒pYA3341-ORF3。将重组质粒电转入鼠伤寒沙门氏菌疫苗株X4550(Asd-、Cya-、Crp-),获得重组疫苗菌株X4550(pYA3341-ORF3)。体外鉴定重组菌表达的GP3蛋白、重组菌特性并进行小鼠免疫试验。酶切鉴定和序列测定证实重组质粒构建成功,SDS-PAGE电泳及Western blot检测有GP3特异性蛋白条带,小鼠免疫后可检测到GP3蛋白抗体。成功构建能稳定表达PRRSV GP3蛋白的口服减毒鼠伤寒沙门氏菌疫苗株。To construct a recombinant attenuated Salmonella typhimurium oral live vector strain expressing the antigen of PRRSV GP3.The PRRSV deleting transmembrane region ORF3 gene was cloned and inserted into expression vector pYA3341（containing Asd）.The recombinant plasmid pYA3341-ORF3 was finally electro-transformed into attenuated S.typhimurium vaccine strains X4550 with delta Asd,Cya and Crp to construct the live vaccine strains X4550（pYA3341-ORF3）.The recombinant strain of X4500（pYA3341-ORF3） was identified its immunogenicity of expression protein,characteristics and immunity evaluation by mouse model.With restriction enzyme digestion and gene sequencing,it was showed that the recombinant plasmid has been constructed successfully.SDS-PAGE and Western blot were used to detect the GP3 protein band and its antigenic property in S.typhimurium X4500（pYA3341-ORF3）.The recombinant strain X4550（pYA3341-ORF3） were immune to mice by gavage,The aimed antibody was detected by ELISA.A recombinant oral attenuated S.typhimurium strain X4500（pYA3341-ORF3） was successfully constructed.

关 键 词：PRRSV GP3 减毒鼠伤寒沙门氏菌 活疫苗 

分 类 号：S858.28[农业科学—临床兽医学]