Figure 3. Tuning TF-biosensors for different contexts.
(a) The TAD and DBD of the TF-biosensor and the corresponding binding site for the DBD in the reporter promoter can be swapped for a different application. Expression of a plasmid-borne luciferase reporter was driven by TF-biosensors containing either a LexA or Gal4 DBD and either a VP16 or B42 TAD. Promoters for the reporter contained DNA-binding sites for either Gal4 or LexA. (b) TF-biosensors were transformed into the yeast strain PJ69-4a and tested for growth on this minimal media containing 1 mM 3-aminotriazole (3-AT) and the indicated steroid. To determine the effect of including an additional destabilization domain, the degron from Matα2 was cloned into one of four positions. (c) G-DIG1-V biosensor response to digoxigenin in yEGFP reporter strain PyE1 either with or without a deletion to the ORF of PDR5. (d) Ligand and TF-biosensor-dependent growth on this media in yeast strains containing deleted ORFs for efflux-related transcription factors (PDR1 and PDR3) or ABC transporter proteins (YOR1, PDR5, SNQ2). In a and c, error bars represent s.e.m. of three biological replicates.