Fig. 1.

(a) Y2H assay to detect ancillary proteins that interact with RdRp. The pGADT7 RdRp was cotransformed in pairs with one of the following clones of pGBK p10/ΔN70pro/VPg/p8 or CP or empty vector into AH109 strain and plated on L, W SD transformation selection plates. The pGAD-RdRp-Vector-pGBK, pGAD-vec-p10-pGBK, pGAD-Vector-ΔN70pro-pGBK and pGAD-Vec-Vec-pGBK were used as negative controls. The pGAD-T antigen-p53-pGBK cotransformant was used as positive control. The summary of Y2H screen results is presented as a table. (b) β-Galactosidase assay to quantitate the strength of interactions: The cotransforments which showed positive interaction were inoculated into quadruple dropout media and grown at 28 °C until the OD₆₀₀ reached to 0.6–1. The assay was performed and β-galactosidase activity units were calculated as described in Section 2. The data is presented as a bar diagram (Y-axis, β-galactosidase activity units (miller units); X-axis Y2H interactions). The pGAD-RdRp-Vector-pGBK, pGAD-Vector-ΔN70pro-pGBK and pGAD-Vector-p10-pGBK (grown in L, W galactose carbon source) were used as negative control. The pGAD-T-antigen-pGBKp53 was used as positive control.