Figure 6
Verification of heterogeneously expressed putative DGCs in E. coli BL21(DE3) using Bc3-5 based dual-fluorescence c-di-GMP reporter.
(a) Verification of putative DGCs by Congo red staining. Strains BLpET and BLpleD were used as negative and positive controls, respectively. (b) Verification of putative DGCs by Bc3-5 based dual-fluorescence c-di-GMP reporter strains. Strains Bc3-5/pET and Bc3-5/pleD were used as negative and positive controls, respectively. Bacterial culture, concentration, resuspension and photography of the concentrated bacterial suspensions were carried out as described in the Methods section. (c) Verification of putative DGCs via RFI measurements. Strains Bc3-5/pET and Bc3-5/pleD were used as negative and positive controls, respectively. Cultures were induced with 1 mM IPTG at 28 °C and RFI values were measured at different induction time points (0–30 h). Significance analysis was conducted from the RFI values of three induction time points (0 h, 2.5 h and 5 h). Data were subjected to one-way analysis of variance (ANOVA) using Bonferroni test with three measurements by OriginLab 8.0 software. *P value < 0.05; **P value < 0.01; ***P value < 0.001; ****P value < 0.0001. Strains used were listed in Table 1. All Data were averages of three independent experiments (error bars were standard deviation from mean values).
