Figure 6.
Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays for Hc-Pro self-interaction. (A) Y2H assay. The yeast cells containing indicated constructs were grown on selective medium lacking leucine, tryptophan and histidine (X-α-Gal). The yeast concentration gradients were 10−1, 10−2 and 10−3, respectively. pGBKT7-p53/pGADT7-RecT (positive control) is symbolized as “+”, and pGBKT7-p53/pGADT7-lamin (negative control) is shown as “–”. (B) BiFC assay of Hc-Pro interactions in N. benthamiana. The full length Hc-Pro and truncated Hc-Pro170 were transiently expressed by co-infiltration on N. benthamiana leaves. sYFPn plus Hc-Pro-sYFPc or Hc-Pro-sYFPn plus sYFPc was used as the negative control. At 3 dpi, YFP fluorescence imaging was performed by confocal laser-scanning microscopy (488 nm). DAPI staining was performed to show the nuclei.