| PRRSV新疆株N基因的克隆及原核表达 |
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| 引用本文： | 王传锋,马伟,张伟,薛英,吴星星,冉多良.PRRSV新疆株N基因的克隆及原核表达[J].新疆农业大学学报,2009,32(6):51-54. |
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| 作者姓名： | 王传锋 马伟 张伟 薛英 吴星星 冉多良 |
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| 作者单位： | 1. 新疆农业大学,动物医学学院,乌鲁木齐,830052
2. 新疆天康畜牧生物技术股份有限公司,乌鲁木齐,830032 |
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| 基金项目： | 新疆农业大学校内前期课题 |
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| 摘 要： | 根据GenBank上已发表的毒株，设计了一对引物扩增PRRSV新疆株N基因片段；将N基因克隆到pMD18-T，在亚克隆到pGEX-6p-1原核表达载体上，构建重组质粒pGEX-6p-N；在37℃条件下，经终浓度为1mmol／L的IPTG诱导表达4h后，获得了可溶性融合蛋白；经SDS-PAGE电泳检测和Western blotting分析，该融合蛋白分子量约为39kDa，与预期结果一致，并能与PRRSV阳性血清发生特异性反应，且非特异性背景很低。
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| 关 键 词： | PRRSV N基因 克隆 原核表达 |
Cloning of Porcine Reproductive and Respiratory Syndrom virus(PRRSV) N Gene and Prokaryotic Expression |
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| WANG Chuan-feng,MA Wei,ZHANG Wei,XUE Yin,WU Xing-xing,RAN Duo-liang.Cloning of Porcine Reproductive and Respiratory Syndrom virus(PRRSV) N Gene and Prokaryotic Expression[J].Journal of Xinjiang Agricultural University,2009,32(6):51-54. |
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| Authors: | WANG Chuan-feng MA Wei ZHANG Wei XUE Yin WU Xing-xing RAN Duo-liang |
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| Institution: | WANG Chuan-feng , MA WeiI , ZHANG WeiI, XUE Yine, WU Xing-xing , RAN Duo-liang(1. College of Animal Medicine, Xinjiang Agricultural University, Urumqi 830052, China ; 2. Xinjiang Tiankang Animal Bio-technology Co. Ltd, Urumqi 830032, China) |
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| Abstract: | The PRRSV nucleoprotein gene amplified by RT-PCR was cloned into pMD18-T vector, and then subcloned into prokaryotic expression pGEX-6p-1, named the recombination plasmids pGEX-6p-N; The IPTG induction with concentration of 1 mmol/L were induced for 4 hours,then the soluble confeitus protein was obtained on the condition of 37 ℃ ,whose molecular weight was obout 39 kDa,which were identical with the predicted results. Western blotting assays suggest that the protein could react with the porcine polyclonal antibodies against PRRSV and proved that recombination N protein has immunological activity. |
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| Keywords: | PRRSV |
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