3.3 PpINH1 interacts with PpVIN2 in Y2H system
Using PpVIN2 as “bait”, each inhibitor protein (“prey”) was tested for its ability to interact with PpVIN2 in the Y2H system. Briefly, a positive result occurs when the bait and prey proteins interact to bring a DNA-binding domain (BD) and activating domain (AD) into proximity, thereby activating the transcription of reporter genes. This enables the yeast host to grow on plates that lack four amino acids, exhibit strong resistance to AbA, and generate blue colonies. Only PpINH1 detectably interacted with PpVIN2 (Figure 3a ). In control experiments (data not shown), yeast cells containing either BD-PpVIN2 or AD-PpINH1 exhibited no self-activation or cell toxicity.