马铃薯S病毒衣壳蛋白表达及多克隆抗体制备  

Prokaryotic Expression and Polyclonal Antibody Preparation of the Potato Virus S Coat Protein

作　　者：王韬远[1] 王永志[2] 张春雨[2] 王忠伟[2] 李闯[2] 张胜利 李小宇[2] WANG Taoyuan;WANG Yongzhi;ZHANG Chunyu;WANG Zhongwei;LI Chuang;ZHANG Shengli;LI Xiaoyu(Wuhu Institute of Technology,Wuhu 241003;Jilin Academy of Agricultural Sciences,Gongzhuling 136100;Jilin Academy of Vegetable and Flower Sciences,Changchun 130000,China)

机构地区：[1]芜湖职业技术学院,安徽芜湖241003 [2]吉林省农业科学院,吉林公主岭136100 [3]吉林省蔬菜花卉科学研究院,长春130000

出　　处：《东北农业科学》2022年第2期30-33,共4页Journal of Northeast Agricultural Sciences

基　　金：安徽省高校自然科学研究重点项目(KJ2019A0981);芜湖职业技术学院重点科学研究项目(Wzyzrzd201906);吉林省农业科技创新工程(CXGC2017TD007)。

摘　　要：马铃薯已成为我国第四大主粮作物,具有重要的战略意义,而马铃薯病毒病是主要的病害之一。本研究通过克隆马铃薯S病毒(potato virus S,PVS)的衣壳蛋白(Coat Protein)基因,连接表达载体pET28b,转化大肠杆菌,体外表达纯化出PVS CP。免疫日本大耳兔,制备出PVS CP多克隆抗体,其识别重组蛋白的效价为64000倍,识别病毒的效价为32000倍;抗体特异性分析表明,制备的多克隆抗体只识别PVS,不识别马铃薯M病毒、马铃薯Y病毒和马铃薯卷叶病毒。PVS多克隆抗体的制备,为马铃薯病毒检测方法提供了技术支持,为种薯质量的提高奠定了基础。The potato has become the fourth major grain crop in China,with important strategic significance,and potato virus disease is one of the main diseases.In this study,the Coat Protein gene of potato virus S(PVS)was cloned,and the expression vector pET28b was ligated into E.coli to express and purify PVS CP.The polyclonal antibody against PVS CP was prepared by immunizing Japanese rabbits.The titer of the recombinant protein was 64,000times and the titer of the recognized virus was 32,000 times.Antibody specificity analysis indicated that the prepared polyclonal antibodies were only recognized by PVS and were not recognized by potato virus M,potato virus Y or potato leaf roll virus.The preparation of PVS polyclonal antibody provides technical support for the detection of potato virus and lays a foundation for the improvement of seed potato quality.

关 键 词：马铃薯S病毒 衣壳蛋白 原核表达 多克隆抗体 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]