Figure 5

Ribo-attenuators allow btuB riboswitch rational re-engineering: (A) Response of each ribo-attenuator to 0, 6.25, and 50 µM 2-aminopurine as compared to the fusion construct (orange). Error bars indicate standard deviation of measurements for biological triplicates. Full single cell data is shown in Supplementary Fig. 6. (B) Fluorescence microscopy of the uninduced and induced btuB fusion and Att2 ribo-attenuated btuB riboswitch, with the latter demonstrating a greater induction difference. White bars represent 1 µm. (C) Plate-killing assay for production of bacteriocin ColE9 from the btuB riboswitch with and without fusion and attenuator domains. Clearances in the E. coli soft agar plate correlate with ColE9 production level, demonstrating a substantially increased protein yield in the ribo-attenuator system.