马铃薯卷叶病毒衣壳蛋白原核表达及多克隆抗体的制备  

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

作　　者：王韬远[1] 张春雨[2] 王忠伟[2] 李闯[2] 张胜利 李建平[2] 李小宇[2] 王永志[2] WANG Taoyuan;ZHANG Chunyu;WANG Zhongwei;LI Chuang;ZHANG Shengli;LI Jianping;LI Xiaoyu;WANG Yongzhi(Wuhu Institute of Technology,Wuhu 241000;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241000 [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》2021年第4期20-23,共4页Journal of Northeast Agricultural Sciences

基　　金：教育部创新发展行动计划(XM-16);芜湖职业技术学院校级重点项目(wzyzrzd201906)。

摘　　要：本研究克隆马铃薯卷叶病毒(Potato leafroll virus,PLRV)衣壳蛋白(Coat Protein, CP)基因,改造其密码子,使其偏好原核表达,构建pCzn1-PLRV CP重组表达载体,通过大肠杆菌表达纯化,经Western Blot方法鉴定为PLRV CP蛋白,纯化后的蛋白免疫日本大耳兔,成功制备PLRV CP蛋白多克隆抗体,识别重组蛋白效价为128 000倍,识别PLRV叶片的效价为32 000倍,经Western Blot方法分析,其特异性良好。本研究为PLRV检测方法的建立及脱毒种薯的检测提供了必需的生物材料。In this study, the potato leafroll virus(PLRV) Coat Protein(CP) gene was cloned, its codon was modified to favor prokaryotic expression, and the pCzn1-PLRV CP recombinant expression vector was constructed, which was expressed and purified by E. coli. The purified protein was identified as PLRV CP protein by Western Blot. The purified protein was immunized with Japanese rabbit, and PLRV CP polyclonal antibody was successfully prepared. The recombinant protein was identified as 128 000 times, and the PLRV leaf was identified as 32 000 times. The specificity was good after Western Blot analysis. This study provides necessary biomaterials for the establishment of PLRV detection method and the detection of virus-free seed potato.

关 键 词：马铃薯卷叶病毒 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]