马铃薯Y 病毒属三种病毒通用型单克隆抗体的鉴定

摘要

Prokaryotic expression vectors pET28-SMV CP,pET28-PVY CP and pET28-CLYVV CP of three Poty-virus viruses (Soybean mosaic virus ,SMV;Potato virus Y ,PVY;and Clover yellow vein virus ,CLYVV)were constructed.Through IPTG induction,expression and purification,we obtained the CP proteins of SMV,PVY and CLYVV.The monoclonal antibodies of 9 strains prepared before were used to detect SMV,PVY and CLYVV viral suspensions and purified CP proteins.The ELISA results showed that the 9 monoclonal antibodies recognized SMV virus;the antibodies 2D3,4F9,4G12 and 6E7 reacted with PVY;CLYVV was recognized by the antibodies 4F9 and 4G12,but the appetency between PVY or CLYVV and antibodies was lower than that of SMV.As for the recombinant expressed CP proteins,SMV and CLYVV CPs reacted with 2D3,4F9,4G12 and 6E7 strongly;PVY CP reacted with 4F9 and 4G12.In general,monoclonal antibodies 4F9 and 4G12 could recognize SMV,CLYVV and PVY universally.This study provided two universal antibodies against Potyvirus .%构建了马铃薯 Y 病毒属的大豆花叶病毒（Soybean mosaic virus ，SMV）、马铃薯 Y 病毒（Potato virus Y， PVY）、三叶草黄脉病毒（Clover yellow vein virus ，CLYVV）的原核表达载体 pET28-SMV CP、pET28-PVY CP 和pET28-CLYVV CP，经 IPTG 诱导、表达和纯化，获得 SMV、PVY 和 CLYVV 的 CP 蛋白。采用 ELISA 方法，用9株实验室制备的单克隆抗体对含 SMV、PVY、CLYVV 的病毒汁液和纯化的重组 CP 蛋白进行检测分析。结果表明，9株单克隆抗体均识别 SMV 病毒，2D3、4F9、4G12和6E7单克隆抗体能够识别 PVY 病毒，4F9和4G12能够识别 CLYVV 病毒，但 PVY、CLYVV 病毒与抗体的亲和力低于 SMV 病毒；对于重组表达的衣壳蛋白：SMV、CLYVV与抗体2D3、4F9、4G12和6E7反应强烈，PVY 与4F9和4G12抗体反应稍强。综上，本研究鉴定出能识别 SMV、PVY、CLYVV 的通用单克隆抗体4F9和4G12。