Figure 2.

Characterization of 5′-OMB and 3′-OMB reporter activities in the context of the pbuE riboswitch. (A) 5′-OMB fluorescence intensity monitored as a function of time by varying the rNTP concentrations to 10, 20, 65 and 100 µM. (B) The 5′-OMB fluorescence emission recorded when transcribing the pbuE riboswitch using either T7 or E. coli RNA polymerase. The fluorescence intensity of the 5′-OMB is represented as a function of the transcription reaction time. (C) The 5′-OMB fluorescence intensity measured at various temperatures in presence of its target sequence. As a control, the 5′-OMB was incubated alone at 45°C and showed no significant fluorescence emission because of temperature destabilization. (D) Effect of DAP concentration on the pbuE riboswitch transcriptional regulation monitored using the 3′-OMB fluorescence emission. DAP was added to the transcription reaction at concentrations ranging from 0 to 25 µM, and the fluorescence emission of the 3′-OMB was measured as a function of time.