Figure 5.

Processing of cspA mRNA by RNase III. (A) Schematic representation of the cspA 5′UTR stem loop. Adapted from Lioliou and co-workers (30). The RNase III processing site is represented by a bold line and the resulting processed mRNA is indicated with an asterisk. (B) cspA^3xFLAG mRNA levels produced from the pCspA^3xFLAG plasmid in the WT, ΔcspA, Δrnc and ΔrncΔcspA strains. MM, Millenium Marker, band lengths are indicated. (C) CspA^3xFLAG protein levels produced from these same strains. Total RNA and protein extraction were performed after growth until mid-exponential phase at 37°C and 200 rpm. RNA samples were run into 1.25% agarose and transferred to Nitran membranes. Northern blots were then developed using a ³²P-labeled anti-FLAG oligo probe and autoradiography. Ethidium bromide staining of 16S RNA is shown as loading control (LC). Protein samples were run into 12% polyacrylamide gels and transferred to nitrocellulose membranes. Western blots were then developed using peroxidase conjugated anti-FLAG antibodies and bioluminescence kit. Coomassie stained gel portions are shown as loading controls (LC).