PDF《江苏农业学报（ Jiangsu J.of Agr.Sci. ） ꎬ2》

江苏农业学报(Jiangsu J.

of Agr.Sci.)?2017?33(5):1099~1104 h ttp: / / www.jsn y x b.com 韩凯凯?李文良?李银?等. 鸭viperin 基因的克隆、表达及 Viperin 蛋白的抗病毒活性[J].江苏农业学报?2017?33(5):1099~1104. doi:10.3969/ j.issn.1000~4440.2017.05.021 鸭viperin 基因的克隆、表达及 Viperin 蛋白的抗病毒 活性 韩凯凯? 李文良? 李银? 毕可然? 刘宇卓? 刘青涛? 赵冬敏? 杨婧? 黄欣梅 (江苏省农业科学院兽医研究所?农业部兽用生物制品工程技术重点实验室?国家兽用生物制品工程技术研究中心?江苏 南京210014) 收稿日期:2017~03~02 基金项目:国家自然科学基金项目(31502101)?江苏省自然科学基 金项目(BK20130710) 作者简介:韩凯凯(1983~)?男?河南新乡人?博士?副研究员?主要从 事家禽病毒分子生物学研究? ( Tel) 025~84390047?( E~ mail)hankk0917@ 126.com 通讯作者:李银?(Tel)025~84391687 摘要: Viperin 蛋白是一种由Ⅰ型和Ⅱ型干扰素诱导的宿主蛋白?广泛存在于各类动物体内?在进化上高度保 守? 本研究以鸭外周血淋巴细胞为原材料?通过 RT~PCR 扩增得到 viperin 基因?同时构建真核表达质粒 pEGFP~N1~ viperin?转染至 DF~1 细胞?用坦布苏病毒攻击后?检测该病毒在转染质粒组和转染空载体组细胞中的增殖情况? 结 果表明?鸭viperin 基因开放阅读框为1

092 bp?编码蛋白由

363 个氨基酸构成? 转染结果显示?viperin 基因在 DF~1 细胞中成功表达? 坦布苏病毒攻击后?与转染空载体组相比?转染 pEGFP~N1~viperin 组显著抑制了坦布苏病毒在 DF~1 细胞中的增殖? 说明鸭 Viperin 蛋白作为抗病毒蛋白?具有良好的抑制坦布苏病毒增殖的作用? 本试验结果 为鸭 Viperin 蛋白抗病毒生物学功能的进一步研究奠定了良好基础? 关键词: viperin 基因? 坦布苏病毒? 抗病毒活性 中图分类号: S858.32 文献标识码: A 文章编号: 1000~4440(2017)05~1099~06 Cloning and expression of duck viperin gene and preliminary study on the antiviral activity of Viperin protein HAN Kai~kai? LI Wen~liang? LI Yin? BI Ke~ran? LIU Yu~zhuo? LIU Qing~tao? ZHAO Dong~min? YANG Jing? HUANG Xin~mei (Institute of Veterinary Medicine? Jiangsu Academy of Agricultural Sciences? Key Laboratory of Veterinary Biologicals Engineering and Technology? Ministry of Agriculture? National Center for Engineering Research of Veterinary Bio~products? Nanjing 210014? China) Abstract: Viperin is a kind of host cellular protein induced by type I and type II interferon? which is found in a wide range of organisms? and is highly conserved in evolution. In this research? we had cloned a full~length open reading frame (ORF) of duck viperin gene from peripheral blood mononuclear cells using RT~PCR. The viperin gene was successful~ ly subcloned into eukaryotic expression vector pEGFP~N1~viperin and transfected into DF~1 cells. After challenged with duck Tembusu virus? the virus replication was analyzed between pEGFP~N1~viperin transfection group and empty vector transfec~ tion group. The results showed that duck viperin gene was amplified and containing

1 092 bp. Transfection results showed that eukaryotic expression vector pEGFP~N1~viperin was successfully expressed in DF~1 cells. Viperin was able to limit the infection of duck Tembusu virus and the virus infection was significant lower than that of the control group. These results indicated that the duck Viperin had well antiviral activity? and it would lay foundations for further research for antiviral biological functions of Viperin. Key words: viperin? Tembusu virus? antiviral ac~ tivity