Figure 4.
A binary vector system for the quantitative analysis of Pdr TF activation by xenobiotics in real time. a, plasmid constructions: Hybrid TFs swapping the natural DNA-binding domain of the Pdr factor with Gal4DBD are constitutively expressed from the pGBKT7 vector. The hybrid factor binds to the GAL1UAS sequence of the pAG413 reporter plasmid driving the expression of lucCP+. Dose-dependent activation of the Pdr TF of choice can be monitorized by the time-elapsed detection of light emission. b, expression of six members of the Pdr TF family of yeast. The Gal4DBD-TF fusion proteins were detected by anti-myc Western blotting in whole cell extracts. The total protein content is visualized by DB71 staining and shown below.