番茄褪绿病毒外壳蛋白多克隆抗体的制备和应用  

Preparation and application of polyclonal antibody against tomato chlorosis virus coat protein

作　　者：伍维兰 聂三妹 张坤[1] 鲁洲 李战彪[2] 季英华[3] 郭灵芳 章松柏[1] WU Wei-lan;NIE San-mei;ZHANG Kun;LU Zhou;LI Zhan-biao;JI Ying-hua;GUO Ling-fang;ZHANG Song-bai(MARA Key Laboratory of Sustainable Crop Production in the Middle Reaches of the Yangtze River(Co-construction by Ministry and Province),Yangtze University,Jingzhou 434025,Hubei,China;MARA Key Laboratory of Green Prevention and Control on Fruits and Vegetables in South China,Guangxi Academy of Agricultural Sciences,Nanning 530007,China;Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]长江大学农业农村部长江中游作物绿色高效生产重点实验室(部省共建),湖北荆州434025 [2]广西壮族自治区农业科学院农业农村部华南果蔬绿色防控重点实验室,南宁530007 [3]江苏省农业科学院植物保护研究所,南京210014

出　　处：《湖北农业科学》2025年第2期197-201,213,共6页Hubei Agricultural Sciences

基　　金：国家自然科学基金面上项目(31972243);广西作物病虫害生物学重点实验室基金项目(22-035-31-22KF03)。

摘　　要：为建立番茄褪绿病毒(Tomato chlorosis virus,ToCV)的快速检测技术,通过RT-PCR技术克隆病毒的主要外壳蛋白(Coat protein,CP)基因,利用Gateway技术将其重组至原核表达载体中,转化至大肠杆菌(Escherichia coli)BL21菌株中诱导表达,将纯化后的CP作为抗原免疫日本大耳兔制备ToCV CP多克隆抗体。间接ELISA显示ToCV CP多克隆抗体的效价为1∶12 800;Western blot和田间试验结果显示多克隆抗体有很好的特异性和灵敏度。ToCV CP多克隆抗体可用于田间ToCV的检测。In order to establish a rapid detection technology for the tomato chlorosis virus(ToCV),the coat protein(CP)gene of the virus was cloned by RT-PCR technology,and it was recombinant into a prokaryotic expression vector using Gateway technology.It was then transformed into Escherichia coli BL21 strain for induction of expression.The purified CP protein was used as an antigen to pre⁃pare a ToCV CP polyclonal antibody by immunizing Japanese rabbits.Indirect ELISA showed that the titer of the ToCV CP polyclonal antibody was 1∶12800.Western blot and field experiments had shown that the ToCV CP polyclonal antibody had good specificity and sensitivity.The ToCV CP polyclonal antibody could be used for the detection of ToCV in the field.

关 键 词：番茄褪绿病毒(ToCV) 外壳蛋白(CP) 原核表达 多克隆抗体 制备 

分 类 号：S432.41[农业科学—植物病理学]