山羊痘病毒L1R蛋白的原核表达及其免疫原性的研究  

Prokaryotic expression and immunogenicity analysis of goatpox virus L1R protein

作　　者：牛贤云 贾怀杰[1] 陈国华[1] 何小兵[1] 张君 房永祥[1] 成温玉[1] 景志忠[1] NIU Xian-yun, JIA Huai-jie, CHEN Guo-hua, HE Xiao-bing, ZHANG Jun, FANG Yong-xiang,CHENG Wen-yu, JING Zhi-zhong(State key laboratory of veterinary Etiological Biology ,Key laboratory of Veterninary Public Health of Ministry of Agriculture ,Lanzhou Veterinary Institute, Chinese Academy of Agricultural Sciences ,Lanzhou 730046, Chin)

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室农业部兽医公共卫生重点实验室

出　　处：《中国兽医科学》2018年第4期450-455,共6页Chinese Veterinary Science

基　　金：“十三五”国家重点研发计划项目(2017YFD0500903,2016YFD0500907);甘肃省农业生物技术专项(GNSW-2012-16)

摘　　要：摘要：为探究山羊痘病毒LIR蛋白的免疫原性，将山羊痘病毒的LIR基因克隆到pET-30a（＋）原核表达载体并转化大肠杆菌BL21（DE3），挑取阳性克隆进行诱导表达。表达产物经SDS-PAGE和Western-blot分析后进行纯化．将纯化的重组L1R蛋白免疫BALB／c小鼠．通过间接ELISA、淋巴细胞增殖和微量中和试验研究重组LIR蛋白的免疫原性及其多克隆抗体的抗病毒作用。结果显示，成功表达出分子质量约为26ku的目的蛋白，其可被山羊抗绵羊痘阳性血清所特异性识别。重组LIR蛋白能够刺激小鼠产生体液免疫和细胞免疫，并且其多克隆抗体能够中和山羊痘病毒，从而证实山羊痘病毒LIR蛋白具有良好的免疫原性。To investigate the immunogenicity of goatpox virus LIR protein,the LIR gene of goatpox virus was cloned into prokaryotic expression vector pET-30a（＋） and transformed into Escherichia coli BL21（DE3） and the positive clone was selected and induced expression. The expressed product was verified by SDS-PAGE and Western-blot and then purified. The BALB/c mice were immuned with the purified recombi- nant LIR protein. The immunogenicity of the recombinant LIR protein and its polyclonal antibody on virus was studied by indirect ELISA,lyphocytes proliferation assay and micro-serum neutralization test. The results showed that the target protein with a molecular weight of about 26 ku was successfully expressed,which could be recognized specially by goat anti-sheeppox positive serum. The recombinant LIR protein could stimulate humoral and cellular immunity in BALB/c mice and its polyclonal antibody could be able to neutralize the goatpox virus. This deminstrates that the LIR protein of goatpox virus possesses good immunogenicity.

关 键 词：山羊痘病毒 LIR蛋白 免疫原性 

分 类 号：S852.659.1[农业科学—基础兽医学]