FIGURE 2.

BdUev1s physically interact with BdUbc13s. (A) Physical interaction between BdUev1s and BdUbc13s in a yeast two-hybrid assay. PJ69-4a cells were transformed with BdUEV1s and BdUBC13s genes and the transformants carrying one Gal4_BD (pGBT9) and one Gal4_AD (pGAD424) plasmid were then selected, replicated onto various plates as indicated and incubated for 3 days before being photographed. The result is representative of at least five independent transformants from each treatment. (B) Protein interactions between BdUev1s and BdUbc13s by an affinity pull-down assay. BL21 (DE3) cells were transformed with pGEX-BdUEV1s, and target gene expression was induced by adding 0.2 mM IPTG. Crude cell extracts were loaded on Glutathione Sepharose^TM 4B beads and 10 μg of purified His₆-BdUbc13.