CVN重组蛋白体外抗猪繁殖与呼吸综合征病毒作用的研究  

Study on effect of recombinant CVN protein agaist porcine reproductive and respiratory syndrome virus in vitro

作　　者：韦应芳 冀池海 汪志[1,2] 盛金良 孙彦阔[1,2] 刘乙兴[1,2] 李琪 蔡孟楷 王衡 张桂红 WEI Ying-fang;JI Chi-hai;WANG Zhi;SHENG Jin-liang;SUN Yan-kuo;LIU Yi-xing;LI Qi;CAI Meng-kai;WANG Heng;ZHANG Gui-hong(College of Veterinary Medicine,South China A gricultural University,Guangzhou 510642,China;Guangdoag Key Laboratory of Zoonosis and Prevention of Zoonosis,Guangzhou 510642,China;Guangdong Provincial Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases,Guangzhou 510642,China;College of Animal Science and Technology,Shihezi University,Shihezi 832003,China)

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]广东省动物源性人兽共患病预防与控制重点实验室,广东广州510642 [3]广东省兽医临床重大疾病综合防控重点实验室,广东广州510642 [4]石河子大学动物科技学院,新疆石河子832003

出　　处：《中国兽医科学》2018年第10期1242-1249,共8页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2018YFD0501200);现代农业技术产业体系基金项目(CARS-35);国家自然科学基金项目(31272564);广东省高等学校优秀青年教师培养计划项目(YQ201530)

摘　　要：本研究以Gen Bank中蓝藻抗病毒蛋白N（CVN）基因序列（DQ668406.1）为基础,合成CVN基因,构建表达质粒pMAL-c5x-CVN,通过优化诱导条件,表达出可溶性CVN重组蛋白。利用cck8试剂盒检测CVN重组蛋白对Marc-145细胞的毒性,并在安全浓度范围内进行了抗猪繁殖与呼吸综合征病毒（PRRSV）试验。在PRRSV感染Marc-145细胞的吸附、穿入和复制三个阶段进行检测,探究CVN体外抗PRRSV的作用效果。各阶段的病毒TCID50检测、间接免疫荧光试验以及qPCR测定结果表明,CVN在PRRSV感染宿主细胞过程中的入侵阶段发挥阻断作用,而在病毒吸附细胞和在细胞内复制的阶段没有抑制效果。本研究为新型抗PRRSV药物的研发奠定了试验基础。A cyanovirus antiviral protein N（CVN） gene was synthesizedbased on the gene sequence（DQ668406.1） in Gen Bank.The soluble recombinant CVN protein was expressed from the CVN expression plasmid,p MAL-c5 x-CVN,by optimizing induction conditions.The cytotoxicity of the recombinant CVN protein on Marc-145 cells was evaluated by cck8 kit.The antiviral effects of CVN on porcine reproduc tive and respiratory syndrome virus（PRRSV）,within a safety concentration range,were assessed via TCID50,indirect immunofluorescence,and q PCR assay during viral adsorption,penetration and replication in the cells.The CVN blocked PRRSV from penetrating into cells,although no inhibitory effect on viral adsorption and replication,providing experimental basis for the development of new anti-PRRSV drugs.

关 键 词：猪繁殖与呼吸综合征病毒 蓝藻抗病毒蛋白N 抗病毒效应 间接免疫荧光试验 

分 类 号：S852.659.6[农业科学—基础兽医学]