FIG. 2.

Western blotting analysis of cells expressing PalI-GFP and PalI^G47D-GFP fusion proteins. Strains expressing wild-type and Gly47Asp PalI-GFP fusion proteins under the control of the alcA^p promoter were cultured overnight in minimal medium with 0.05% (wt/vol) glucose and shifted for 3 h to either 1% ethanol-containing medium (I, inducing conditions for alcA^p) or 1% glucose-containing medium (R, repressing conditions for alcA^p) before proceeding to membrane protein extraction. Proteins were analyzed by Western blotting, which was reacted using a cocktail of monoclonal anti-GFP antibodies (Roche). Approximately equal loading of the different lanes was confirmed after protein staining of a duplicate gel. Standards (at left) are in kDa.