| 葡萄A病毒外壳蛋白原核表达及抗血清制备 |
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| 引用本文： | 任 芳,董雅凤,张尊平,范旭东,胡国君,朱红娟.葡萄A病毒外壳蛋白原核表达及抗血清制备[J].植物病理学报,2014,44(3):327-331. |
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| 作者姓名： | 任 芳 董雅凤 张尊平 范旭东 胡国君 朱红娟 |
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| 作者单位： | 中国农业科学院果树研究所，国家落叶果树脱毒中心，兴城 125100 |
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| 基金项目： | 国家葡萄产业技术体系建设项目(CARS\|30\|bc\|3) |
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| 摘 要： | 葡萄A病毒（Grapevine virus A，GVA）为线性病毒科（Flexiviridae）葡萄病毒属（Vitivirus）的代表种，是葡萄皱木复合病（rugose wood complex disease）的重要病原之一，可引起葡萄嫁接成活率下降、春季萌芽延迟、生长减弱甚至衰退死亡等危害\1，2\]。GVA为线状单链RNA病毒，基因组共编码5个开放阅读框（ORF1\|5），其中ORF4 编码外壳蛋白（coat protein， CP），是病毒粒子包裹和系统移动所必需的功能蛋白\3，4\]。GVA自然寄主为葡萄，机械摩擦可侵染本氏烟等草本寄主\2\]，由于嫁接和无性繁殖材料调运等因素造成该病毒远距离传播，目前在世界多个国家和地区均有发生。
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| 收稿时间： | 2013-08-30 |
Prokaryotic expression of Grapevine virus A coat protein and antiserum preparation |
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| REN Fang,DONG Ya\|feng,ZHANG Zun\|ping,FAN Xu\|dong,HU Guo\|jun,ZHU Hong\|juan.Prokaryotic expression of Grapevine virus A coat protein and antiserum preparation[J].Acta Phytopathologica Sinica,2014,44(3):327-331. |
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| Authors: | REN Fang DONG Ya\|feng ZHANG Zun\|ping FAN Xu\|dong HU Guo\|jun ZHU Hong\|juan |
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| Institution: | National Center for Production Virus\|free Deciduous Fruit Tree, Research Institute of Pomology, Chinese Academy of Agriculture Sciences, Xingcheng 125100, China |
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| Abstract: | In order to develop serological diagnostic tools for rapid detection of Grapevine virus A (GVA), the coat protein gene of GVA isolate LN68 (JF754571) was inserted into expression vector pET\|28a. The recombinant plasmid pET\|GVA CP was then transformed into E. coli BL21 (DE3) plysS strain. An approximately 26 kDa recombinant GVA CP protein was induced at a high level by IPTG in SDS\|PAGE analysis. The purified protein was used as antigen for raising GVA CP antiserum in rabbits. The specificity of antiserum of GVA was tested by Western blot and ELISA. It was shown that the antiserum could be successfully used to detect the recombinant protein and the purified GVA coat protein in Western blot analysis, and also in indirect\|ELISA to detect GVA in grapevine and the infected Gomphrena globos at dilutions of up to 1:2 000. Large amounts of the target protein with high antigenicity could be yielded by expression of CP gene, and antiserum obtained in the study could be used for specific detection of GVA. |
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| Keywords: | Grapevine virus A coat protein prokaryotic expression antiserum preparation |
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