| 黄瓜绿斑驳花叶病毒cp基因的原核表达及抗血清制备 |
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| 引用本文： | 陈红运,赵文军,白静,李桂芬,朱水芳.黄瓜绿斑驳花叶病毒cp基因的原核表达及抗血清制备[J].植物病理学报,2007,37(5):467-471. |
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| 作者姓名： | 陈红运 赵文军 白静 李桂芬 朱水芳 |
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| 作者单位： | 1 中国检验检疫科学研究院动植物检疫研究所 北京 100029;2 沈阳农业大学植物保护学院 沈阳 110161 |
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| 基金项目： | 国家质检总局资助项目(2007IK253) |
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| 摘 要： | 采用RT-PCR方法克隆了黄瓜绿斑驳花叶病毒辽宁分离物(Cucumber green mottle mosaic virus Liaoning isolate, CGMMV-LN)的cp基因并连接到原核表达载体pGEX-4T-3和pET-22b (+)上, 将获得的重组子pGEX-4T-3-CGMMV CP和pET-22b (+)-CGMMV CP转化大肠杆菌BL21后用IPTG进行诱导表达。SDS-PAGE和W estern blot分析表明, cp基因在大肠杆菌中获得了高效表达, 融合蛋白分子量分别为43.8 kDa和17.3 kDa。将17.3 kDa融合蛋白纯化后免疫家兔, 制备了CGMMV特异性抗血清, 抗原包被间接ELISA法(ACP-ELISA)测定抗血清的效价为1/20 000。
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| 关 键 词： | 黄瓜绿斑驳花叶病毒 外壳蛋白 原核表达 抗血清 ACP-ELISA |
| 文章编号： | 0412-0914（2007）05-0467-05 |
| 修稿时间： | 2006-07-23 |
Expression of cp gene and antiserum preparation of Cucumber green mottle mosaic virus |
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| CHEN Hong-yun, ZHAO Wen-jun , BAI Jing, LI Gui-fen, ZHU Shui-fang.Expression of cp gene and antiserum preparation of Cucumber green mottle mosaic virus[J].Acta Phytopathologica Sinica,2007,37(5):467-471. |
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| Authors: | CHEN Hong-yun ZHAO Wen-jun BAI Jing LI Gui-fen ZHU Shui-fang |
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| Affiliation: | 1 Institute of Animal and Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100029, China;2 College of Plant Protection, Shenyang Agricultural University, Shenyang 110161, China |
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| Abstract: | The coat protein(CP) gene of Cucumber green mottle mosaic virus Liaoning isolate(CGMMV-LN) was amplified by RT-PCR,and ligated to the expression vectors pGEX-4T-3 and pET-22b( ).The recombinant plasmids pGEX-4T-3-CGMMV CP and pET-22b( )-CGMMV CP were transformed into E.coli BL21 respectively and then induced by IPTG.It was shown that the cp gene was highly expressed by SDS-PAGE and Western blot analysis.The molecular weights of recombinant proteins were 43.8 kDa and 17.3 kDa in pGEX-4T-3 and pET-22b( ) vectors.Antiserum with high specificity was produced after the rabbit was immunized with 17.3 kDa purified recombinant protein,and the titer was 1/20 000 detected by antigen coating plate ELISA(ACP-ELISA). |
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| Keywords: | Cucumber green mottle mosaic virus CP prokaryotic expression antiserum ACP-ELISA |
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