Fig. 1: Activity of misfolded protein biosensor and mCherry fusion proteins.
From: Biosensor Guided Polyketide Synthases Engineering for Optimization of Domain Exchange Boundaries
a GFP fluorescence of E. coli biosensor strains expressing PKSs with variable solubilities. pET = pET28a (empty vector control). b Fluorescence of mCherry tagged PKSs (left y-axis) and SDS-PAGE quantified abundance of the proteins relative to DEBSM6 amount (right y-axis). (c) mCherry fluorescence and (d) SDS-PAGE quantification of PKS proteins in different protein fractions of lysed cells, relative to “Total protein” for each replicate. Cells were induced with 250 µM IPTG in (a–d). e Fluorescence of ΔarsB::Pibp GFP strain expressing mCherry tagged PKSs. f Same results as 1e with a simplified “solubility coefficient”: the ratio of the expressed protein (mCherry fluorescence) over activation of insolubility biosensor (GFP fluorescence). Data is presented as mean values of three biological replicates, dots are individual data points. In (a–c) fluorescence was measured using a microplate spectrophotometer, whereas in (e) fluorescence was measured using flow cytometry. Auto Fl = auto fluorescence, arb. units = arbitrary units. Source data are provided as a Source Data file.
