Fig. 4.

RYSV N specifically interacted with P in vitro. a-I Western blot results of GST pull-down assay to detect the interaction of N with P. Bait protein pGEX-3X-N or pGEX-3X was incubated with prey protein pDEST17-P. Antibody against GST was used to detect GST-tagged protein; His antibody was used to detect the bound proteins. The interaction of N with N (a-II) and P with P (a-III) was also verified by GST pull-down assay. b Yeast two-hybrid assay for the interaction of N with P, N with N and P with P. Yeasts transformed with the bait and prey constructs, were grown on SD/−Trp/−Leu/-His/−Ade medium supplemented with 40 μg/ml X-α-Gal. pGBKT7–53 and pGADT7-T co-transformant was used as the positive control. Yeast cells co-transformed with pGBKT7-Lam/ pGADT7-T, pGBKT7-N/pGADT7 and pGBKT7/pGADT7-P were used as negative control