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小麦矮缩病毒(Wheat dwarf virus,WDV)引起的小麦矮缩病是近年来我国小麦生产中的一种重要病毒病害,急需研发快速精准的检测技术用于预测预报和病毒-介体相互作用的研究。本研究应用Gateway重组技术构建了外壳蛋白基因(Coat protein,CP)的原核表达载体,将重组表达载体转化大肠杆菌Rosetta,经IPTG诱导获得CP基因原核表达蛋白。以重组蛋白为抗原免疫新西兰大白兔制备得到了相应的抗体,Western blot检测表明制备的抗体能与CP重组蛋白、感病小麦和带毒叶蝉特异性结合,说明获得的抗体特异性高。用获得的抗体进行免疫荧光标记,观察到病毒分布在介体叶蝉的前中肠和中中肠部位,为WDV的预测预报和介体条沙叶蝉传毒机制的研究奠定了基础。
Wheat dwarf virus caused by wheat dwarf virus (WDV) is an important viral disease in wheat production in China in recent years. It is urgent to develop rapid and accurate detection techniques for prediction and virus-mediator interaction the study. In this study, we constructed a prokaryotic expression vector of Coat protein (CP) using Gateway recombination technology. The recombinant expression vector was transformed into E.coli Rosetta, and prokaryotic expression of CP gene was induced by IPTG. The recombinant antibodies were used to immunize New Zealand white rabbits to prepare the corresponding antibodies. The results of Western blot showed that the antibodies could specifically bind to CP recombinant protein, susceptible wheat and infected leafhopper, indicating that the obtained antibody is highly specific. Immunofluorescent labeling of the obtained antibodies was used to observe the distribution of the virus in the middle and mid intestine of the leafhoppers, which laid the foundation for the prediction and prediction of WDV and the mechanism of drug-mediated transmission.