Fig. 4
From: Evaluation of GFP reporter utility for analysis of transcriptional slippage during gene expression
Analysis of polyA- and polyT-based gfp mRNA levels and stability. a Top panel: total mRNA was extracted from E. coli ER2566 bearing pBADmingfpA60, pBADmingfpT60 and pETmingfpA60 plasmids after 10 min (lanes 1–3) and 30 min expression (lanes 4–6), respectively, generated by E. coli (lanes 1, 2, 4 and 5) or T7 phage RNAP (lanes 3 and 6). The membrane was probed with biotin labelled gfp DNA. Lane 7, no-gfp control bacteria. Arrow—prominent degradation product. Bottom panel: ethidium bromide-stained 16S rRNA are shown as loading control. b gfp transcripts stability. Rifampicin was added to an exponential culture of E. coli ER2655 growing in LB medium after 10 min of 0.1% L-arabinose induction. The mRNA levels were determined by RT-qPCR, using stable 16S rRNA as the internal standard. The circles represent gfpA60 and triangles represent gfpT60 transcripts. All mRNA levels were normalized to 1 at time = 0 (the points overlap). The data were fitted to an exponential decay. Standard error bars from three determinations are shown