Fig. 4.

Mapping of p10 interacting region on RdRp using Y2H assay. (a) RdRp and its C-terminal deletion mutants (pGAD-RdRpCΔ43 and pGAD-RdRpCΔ85) were cotransformed with pGBK p10 and plated on L⁻ and W⁻ transformation selection plates. The colonies obtained were restreaked on fresh L⁻ & W⁻ plates (column 1). The cells grown on L⁻ & W⁻ plates were replica plated on interaction selection plates [(L⁻, W⁻ & H⁻ + 5 mM 3AT (column 2); L⁻, W⁻, H⁻ & Ade⁻ (column 3)]. (b) Quantitation of interaction by β-Galactosidase assay: The β-galactosidase assay was carried out with cells grown on L⁻, W⁻, H⁻ and Ade⁻ plates. The results obtained are presented as a bar diagram (Y-axis, β-galactosidase activity units; X-axis Y2H interactions). The pGAD-T-antigen-p53-pGBK was used as positive control and pGAD-RdRp-Vector-pGBK and pGAD-Vector-p10-pGBK were used as negative control (grown on L⁻ & W⁻ galactose media). The pGAD-RdRpCΔ85-p10-pGBK was also grown in L⁻ & W⁻ galactose media.