Fig. 4: Alternative strategies to engineer the pBAD promoter.
a The pBAD promoter was mutated between the −35 and −10 boxes25. The pBAD promoter in the base strain (Fig. 1a) was replaced with the mutated pBAD promoter and cultured in 96-well plates. Growth was measured in a plate reader at different induction levels of GPD1 (0, 0.1, 0.3, 0.5, 1, and 2% ara). Glycerol in the medium was measured after 24 h. Growth rates were determined by regression analysis between 5 and 10 h. Small dots show data from n = 2 plate reader cultures and big dots are the mean. b Same as a, for strains expressing gpd1 with engineered pBAD promoters that have 0 to 3 Cra-binding sites inserted at different positions. The position of Cra-binding sites is indicated in orange. c GFP plasmids with or without Cra-binding site were expressed in the wild-type (WT) or the Δcra strain. (n = 2, pBAD, WT is the same as in Fig. 1b). Source data are provided in the Source Data file.
