| 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 |
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| 引用本文： | 方剑玉,朱文豪,郭小参,白献晓,李海利,徐引弟,郎利敏,王治方,王克领.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). |
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| 作者姓名： | 方剑玉 朱文豪 郭小参 白献晓 李海利 徐引弟 郎利敏 王治方 王克领 |
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| 作者单位： | 1. 河南省农业科学院 畜牧兽医研究所,河南 郑州 450002;南京农业大学 动物医学院, 江苏 南京 210095;2. 河南省农业科学院 畜牧兽医研究所,河南 郑州,450002;3. 普莱柯生物工程股份有限公司,河南 洛阳,471000 |
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| 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RTPCR方法扩增猪Viperin基因,并克隆入腺病毒穿梭载体p Ad Track-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(p Ad Track-s VIP)电转化大肠杆菌BJ5183,与腺病毒骨架载体p Ad Easy-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r Ad-s VIP。重组腺病毒r Ad-s VIP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r Ad-s VIP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。
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| 关 键 词： | 猪Viperin蛋白 重组腺病毒 抗病毒活性 猪繁殖与呼吸综合征病毒 |
Expression of Swine Viperin Protein in Recombinant Adenovirus and Study of Its Effect on PRRSV Replication |
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| FANG Jianyu,ZHU Wenhao,GUO Xiaocan,BAI Xianxiao,LI Haili,XU Yindi,LANG Limin,WANG Zhifang,WANG Keling.Expression of Swine Viperin Protein in Recombinant Adenovirus and Study of Its Effect on PRRSV Replication[J].Journal of Henan Agricultural Sciences,2017,46(9). |
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| Authors: | FANG Jianyu ZHU Wenhao GUO Xiaocan BAI Xianxiao LI Haili XU Yindi LANG Limin WANG Zhifang WANG Keling |
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| Abstract: | To study the inhibitory effect of swine Viperin on PRRSV replication,the swine Viperin gene was amplified by RT-PCR and cloned into the adenovirus shuttle vector pAdTrack-CMV.The recombinant plasmid were identified by enzyme digestion and sequencing analysis.The pAdTrack-sVIP was linearized by Pme Ⅰ and homologous recombined with pAdEasy-1 backbone vector in E.coli BJ5183.The recombinant adenoviral genome DNA was extracted and digested by Pac Ⅰ,then the DNA was transfected into the HEK-293A cells for packaging whole virus particle rAd-sVIP.The CPE and GFP fluorescence of cells were observed,the cells were harvested and adenoviral expressed recombinant swine Viperin protein were identified by RT-PCR and Western blot.The result showed that the swine Viperin was correctly expressed as expected.The expressed swine Viperin protein had good reactivity,and it could inhibit the PRRSV replication. |
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| Keywords: | swine Viperin protein recombinant adenovirus antiviral activity PRRSV |
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