南方水稻黑条矮缩病毒P6抗体的制备及应用

Preparation and Application of Polyclonal Antibodies Against P6 Protein of Southern Rice Black Streaked Dwarf Virus

为了建立快速准确检测田间介体白背飞虱带毒率的方法,利用Gateway重组技术将与病毒早期复制有关的P6基因的N端993 bp构建原核表达载体pDEST17-P6,并将诱导表达的目的蛋白免疫注射新西兰大白兔制备多克隆抗体。结果表明:Western blot检测制备的P6抗体可特异性检测SRBSDV侵染水稻的P6蛋白,免疫荧光标记技术可检测南方水稻黑条矮缩病毒(Southern rice black streaked dwarf virus,SRBSDV)侵染白背飞虱体内的P6蛋白,且P6抗体可直观高效准确地用于免疫荧光标记检测昆虫带毒率。以上研究表明,所制备的P6抗体可用于SRBSDV的田间介体昆虫带毒率快速检测,有助于病毒病害的监测和预报。

To establish a method that allowing fast and efficient detection of SRBSDV, a fragment comprising the N-terminal 993 bp of P6 ORF of SRBSDV was cloned into the vector pDEST17 using Gateway recombination technology. Antiserum was obtained by injecting the bacterially expressed peptide to healthy rabbits. Western blot experiments showed that the antiserum could detect P6 specifically in rice plants infected by SRBSDV. The fluorescence antibody could also be used to detect P6 in WBPHs using immunofluorescent labeling technique. These results indicated that P6 antibodies prepared here could be used for efficient and accurate detection of SRBSDV in diseased rice plants and viruliferous vectors by immunofluorescent labeling techniques, which was important for the monitoring and forecasting of SRBSDV occurrence.