Fig. 7: Translation enhancement pattern is observed in E. coli.

a Schematic of the dual-color fluorescence reporter system. The two reporters, mCherry and GFP, have the same T7 promoter (yellow) and ribosome binding site (RBS in orange). The 5′UTR of the GFP contains the 10 hp hairpin and potential G-quadruplex sequence (PQS in red) that generates RNA G-quadruplex (RG4). b Fluorescence imaging of GFP expression in E. coli. The scale bar indicates 6 μm (50 px). The images shown are one representative result from n = 3 independent experiments. c, d Real-time GFP and mCherry intensities. The curves represent example traces of cMyc, where non-template (NT), control (C), and template (T) are colored in blue, gray, and purple, respectively. The data is collected after IPTG induction by plate reader. The curves shown are one representative result from n = 3 independent experiments. c The orientation-dependence is observed in GFP. d The three curves show no difference in mCherry. e Normalized translation efficiencies are defined by the ratio of GFP and mCherry signal at 210 min after induction. The PQS-NT constructs showed higher translation than T and the translation increased with bulky PQS. The calculation is described in “Method” and Supplementary Fig. 5. Data are presented as mean ± SEM of n = 3 independent experiments. Shown in (e) only represents the significance between template and non-template, where *P < 0.05 (two-sided paired t-test). Exact mean values are provided in Supplementary Table 3.10. Raw data points are provided as a Source Data file.