表达猪Viperin蛋白重组腺病毒的构建及其对PRRSV复制的抑制作用

河南农业科学 ›› 2017, Vol. 46 ›› Issue (9): 126-131.

表达猪Viperin蛋白重组腺病毒的构建及其对PRRSV复制的抑制作用

方剑玉^1,2，朱文豪¹，郭小参³，白献晓¹，李海利¹，徐引弟¹，郎利敏¹，王治方¹，

1.河南省农业科学院畜牧兽医研究所，河南郑州 450002； 2.南京农业大学动物医学院，江苏南京 210095； 3.普莱柯生物工程股份有限公司，河南洛阳 471000

收稿日期:2017-04-06 出版日期:2017-09-15 发布日期:2017-09-15
通讯作者: 王克领（1964-），男，河南柘城人，研究员，本科，主要从事动物传染病研究。E-mail:1449578205@qq.com
作者简介:方剑玉（1982-），女，河南林州人，助理研究员，博士，主要从事兽医免疫学研究。E-mail:fangjianyu2006@126.com
基金资助:
国家自然科学基金项目（31672565）

Received:2017-04-06 Published:2017-09-15 Online:2017-09-15

摘要/Abstract

摘要： 为研究猪Viperin蛋白对猪繁殖与呼吸综合征病毒（PRRSV）复制的抑制作用，通过RT-PCR方法扩增猪Viperin基因，并克隆入腺病毒穿梭载体pAdTrack-CMV，经菌液PCR、酶切鉴定后进行测序。将PmeⅠ内切酶线性化的重组穿梭载体质粒（pAdTrack-sVIP）电转化大肠杆菌BJ5183，与腺病毒骨架载体pAdEasy-1进行同源重组，提取重组后的腺病毒质粒，经内切酶PacⅠ线性化后转染HEK-293A细胞，装配成完整的病毒粒子rAd-sVIP。重组腺病毒rAd-sVIP感染细胞后，可见绿色荧光蛋白的表达。RT-PCR、Western blot检测结果表明，重组腺病毒rAd-sVIP可正确表达猪Viperin蛋白，且表达的猪Viperin蛋白具有良好的反应原性，对PRRSV的复制具有明显的抑制作用。

方剑玉，朱文豪，郭小参，白献晓，李海利，徐引弟，郎利敏，王治方. 表达猪Viperin蛋白重组腺病毒的构建及其对PRRSV复制的抑制作用[J]. 河南农业科学, 2017, 46(9): 126-131.

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