Fig. 2.

Design and expression of two transcription factor-based biosensors in V. natriegens A) Schematic overview of LysG and Lrp biosensor design for expression in V. natriegens. Different sensor variants were created using promoters from the Anderson promoter library to express lrp or lysG. The first part of brnF or lysG were added to the respective construct, because these genes are transcribed as a leaderless transcripts. After the first part of brnF/lysE, a stop codon (pin) and an additional RBS (bubble) were added (for further details, see supplementary data 2 and 3). B) Sensor response of the Lrp and LysG based biosensor variants. Strains were induced with 3 mM of the indicated inducer, or no inducer was added. C) Dose-reponse signal of the LysG based biosensor variants and L-lysine. A hill-curve was fitted to the measured values. For all fluorescence measurements, fluorescence per cell of 10,000 cells per culture were measured and averaged, and fold change over background was calculated. Average and standard deviation of three cultures are shown (n = 3). Promoters from the Anderson promoter library are shown as abbreviations (P100 = J23100, etc.).