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. Author manuscript; available in PMC: 2015 Jul 31.
Published in final edited form as: J Am Chem Soc. 2015 May 15;137(20):6432–6435. doi: 10.1021/jacs.5b00275

Figure 3.

Figure 3

An in vivo activity assay for novel diadenylatecyclases using cyclic dinucleotide-selective biosensors. (a) Schematic of the two plasmid screening system. E. coli is co-transformed with a biosensor-encoding plasmid and an enzyme-encoding plasmid. After induction of expression, cells are incubated with DFHBI-1T and fluorescence is analyzed. (b) Flow cytometry data for biosensors upon coexpression with no enzyme or a known diguanylate cyclase (WspR), cyclic AMP-GMP synthase (DncV), or diadenylate cyclase (DisA). (c) Flow cytometry data for biosensors upon coexpression with predicted diadenylatecyclases from M. jannaschii and C. difficile. Fluorescence values from (b) and (c) are the average of three independent biological replicates and are shown with standard deviations in Table S1. (d) Representative flow cytometry graph for the yuaA-Spinach2 biosensor using logicle scaling on x-axis.