鸭抗病毒蛋白(viperin)在转染的BHK-21细胞表达并抑制鸭坦布苏病毒出芽  

Duck viperin can express in transfected BHK-21 cells and inhibit germination of Duck Tembusu virus

作　　者：周祺[1,2] 顾香雪 许泽军 黄荣 殷冬冬 萨达娅[1] 吕炫 王桂军 ZHOU Qi;GU Xiangxue;XU Zejun;HUANG Rong;YIN Dongdong;AYAZ Sada;LV Xuan;WANG Guijun(College of Animal Science and Technology,Anhui Agricultural University;Anhui Provincial Key Laboratory of Veterinary Pathobiology and Disease Control,Hefei 230036,China)

机构地区：[1]安徽农业大学动物科技学院 [2]兽医病理生物学与疫病防控安徽省重点实验室,安徽合肥230036

出　　处：《细胞与分子免疫学杂志》2019年第5期412-418,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：科技部国家重点研发计划(2018YFD0500100,2016YFD0500805);安徽农业大学研究生创新基金(2018yjs-22);兽医病理生物学与疫病防控安徽省重点实验室自主课题(SYZZ2017003)

摘　　要：目的探索鸭抗病毒白(viperin)对鸭坦布苏病毒(DTMUV)增殖的影响。方法使用生物学信息学方法分析鸭viperin基因,构建系统发育树。构建原核表达质粒pGEX-6P-viperin和真核表达质粒pEGFP-N1-viperin,分别转化Rosseta感受态细胞和转染BHK-21细胞。用DTMUV分别感染真核表达质粒和空载体质粒的BHK-21细胞,利用实时荧光定量PCR检测细胞沉淀和上清中DTMUV的含量,利用质谱的方法鉴定增强型绿色荧光蛋白(EGFP)单克隆抗体捕获的结合蛋白。结果鸭viperin基因与其他种属viperin基因有显著差异。鸭viperin蛋白能成功在BHK-21细胞表达,鸭viperin抑制DTMUV在BHK-21细胞中的出芽。质谱分析结果显示转染真核表达质粒的细胞中可能存在6种影响抑制DTMUV增殖、出芽的蛋白。结论鸭viperin蛋白可在BHK-21细胞表达,且能抑制鸭坦布苏病毒出芽。Objective To measure the effect of duck viperin protein on the proliferation of duck Tembusu virus(DTMUV). Methods We analyzed the duck viperin gene using a bioinformatics. Plasmids pGEX-6P-viperin and pEGFP-N1-viperin were constructed and transformed into Rosseta and transfected into BHK-21 cells, respectively. BHK-21 cells transfected with plasmid pEGFP-N1-viperin and empty vector were infected with DTMUV. The content of DTMUV in cell precipitation and supernatant was analyzed by real-time fluorescent quantitative PCR. Finally, the effect of viperin on DTMUV proliferation and the binding proteins captured by EGFP monoclonal antibody were evaluated using mass spectrometry. Results Significant differences were observed between the expression levels of viperin gene in duck compared to other species. Duck viperin was found to inhibit the budding of DTMUV in BHK-21 cells. Further, we identified six proteins that might be involved in the inhibition of the proliferation and budding of DTMUV, possibly indicative of the viperin anti-Tembusu virus pathway in ducks. Conclusion Expression patterns of duck viperin reveal how the budding of duck Tembusu virus is inhibited.

关 键 词：鸭 viperin蛋白 鸭坦布苏病毒(DTMUV) 质谱分析法 

分 类 号：R392.1[医药卫生—免疫学] R39233[医药卫生—基础医学]