马铃薯Y病毒属三种病毒通用型单克隆抗体的鉴定

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。

Identification of universal monoclonal antibodies against three viruses of Potyvirus

Prokaryotic expression vectors pET28-SMV CP, pET28-PVY CP and pET28-CLYVV CP of three Potyvirus 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.