褐飞虱flightin基因多克隆抗体的制备及应用

he brown planthopper (BPH; Nilaparvata lugens Stal), a kind of migratory insects, is the primary pests of rice in Asian countries. Previous studies on flightin gene indicated that it is playing a crucial role in the process of BPH wing dimorphism. To determine the expression difference of flightin protein in different wing forms and different instar of BPHs, a prokaryotic expression vector pDEST17-flight was constructed through the Gateway recombination technology. Anti-flightin polyclonal antibody was obtained from the antisera of New Zealand white rabbit, which was injected with IPTG-induced protein as antigen. The working concentration of IPTG was 1mmol/L. Western blotting analysis indicated that the flightin antibodies can specifically react with flightin protein in BPH. The ELISA assay showed that the potency of antiserum was 1:6400. Strong expression of flightin protein was detected in macropterous BPH adults and trace expression of flightin protein was detected in brachypterous BPH male ones, but no expression was detected in the brachypterous female ones and nymph ones.