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. 2017 Jan 26;8(2):50. doi: 10.3390/genes8020050

Figure 9.

Figure 9

Sequences within the ryfA1 transcript do not mediate translation initiation. (a) Western blot analysis was used to detect GFP production in E. coli carrying a negative control plasmid (pXG-0), the experimental plasmid p5′UTR RyfA1, or a positive control plasmid (pXG-1). Under the conditions tested, no GFP was produced in the strain carrying p5′UTR RyfA1, indicating that the cloned sequences do not support translation initiation, and thus that the ryfA1 transcript likely does not encode a small peptide. (b) qRT-PCR was used to ensure that the same triplicate samples used to detect GFP protein were producing gfp transcript. Strains carrying either the positive control or the p5′UTR RyfA1 plasmid produce significantly more transcript than the negative control. Error bars represent one standard deviation. * indicates a significant difference (p < 0.05). Relative levels were calculated using the ΔΔCt method [25] where gfp levels are normalized to the amount of rrsA present in each sample and are expressed relative to that present in a single pXG-10 sample.