Fig. 2
From: Evaluation of GFP reporter utility for analysis of transcriptional slippage during gene expression
Evaluation of frameshifted (− 1) gfp fusion’s usefulness to serve as transcriptional slippage reporter. a Details of the nucleotide sequences of the proximal part of gfp fusions. The names of plasmid constructs’ and reporter gene variants, nucleotide sequences with possible sites of insertion slippage events (indicated by arrows), and primary and slippage-induced amino acid sequences are shown. Reading frame of genes (− 1 or 0) is reflected in their names as suffix 0 or − 1, respectively. Actual (black, below) and native (green, above) amino acid numbering of the GFPA60 hybrid is provided. b Relative fluorescence level of indicated GFP hybrids. All measurements were performed in three to five duplicate repetitions. Error bars represent standard deviations. c Western blotting of the total cell extracts of hybrids shown in b and GFP immunodetection with ECL chemiluminescence system. d Extraordinary slippage properties of T7 RNAP in contrast to E. coli RNAP. ER2566 cells with appropriate pET24a-(lanes 1–5) or pBAD24-derived plasmids (lanes 7–10) carrying − 1 frameshifted gfp genes were induced with 1 mM IPTG or 0.1% l-arabinose at 37 °C, respectively. Aliquot of cell extracts were western blotted and immunodetected with anti-GFP primary antibodies. Below the molecular weight of the GFP protein products in kDa are provided