山羊痘病毒ORF112蛋白的原核表达及其免疫原性分析  

Prokaryotic Expression and Immunogenicity Analysis of ORF112 Protein of Goatpox Virus

作　　者：陈伯祥[1] 杨明[1] 成伟伟[1] 李杰[1] 豆林涛[1] CHEN Bo-xiang;YANG Ming;CHENG Wei-wei;LI Jie;DOU Lin＇tao(Gansu Institute of Animal and Veterinary Science,Pingliang,Gansu,744000,China)

机构地区：[1]甘肃省畜牧兽医研究所,甘肃平凉744000

出　　处：《动物医学进展》2018年第9期5-9,共5页Progress In Veterinary Medicine

基　　金：甘肃省科技支撑计划项目(1104NKCL097)

摘　　要：为表达山羊痘病毒ORF112蛋白并研究其免疫原性,构建了含有ORF112基因的重组表达质粒pET-ORF112;诱导表达和纯化了ORF112蛋白,SDS-PAGE和Western blot对该蛋白进行了分析和生物活性鉴定,并用分析鉴定后的ORF112蛋白免疫接种Balb/c小鼠,采集小鼠血清用间接ELISA进行了抗体检测。结果显示,表达和纯化了含有GST标签的GST-ORF112蛋白,大小约23ku,Western blot显示ORF112蛋白具有较好的特异性,间接ELISA抗体检测表明ORF112蛋白具有良好的免疫原性,为深入研究ORF112蛋白的生物学功能及对山羊痘的诊断与防控奠定了基础。To study the immunogenicity and expression of ORF112 protein from goatpox virus, ORF112 gene was amplified from goatpox virus and cloned into pET-28a, and the recombinant plasmid was named as pET-ORF112. The recombinant ORFt112 protein was expressed in Escherichia coli by induction of IPTG （1 mmol/L）,and the expressed ORF112 protein was purified by affinity chromatography.This protein was analyzed by SDA-PAGE,and biological activity was determined by Western blot.Balb/c mice were immu- nized with the purified ORF112 protein to produce immune serum.Polyclonal antibody from immune serum was detected by ELISA.The results indicated that the expressed ORF112 protein, with about 23 ku molecu- lar weight,showed a good antibody specificity by Western blot and a good immunogenicity by ELISA.The result laid the solid foundation for the further study on biological function of ORF112 protein and diagnosis and prevention of goatpox.

关 键 词：山羊痘病毒 ORF112蛋白 免疫原性 

分 类 号：S852.659.1[农业科学—基础兽医学] Q786[农业科学—兽医学]