Figure 3.
Differential termination strengths of rho-independent terminators (RITs) at different transcription levels. (A) Design of the dual reporter assay plasmid, pDRA2, with transcription initiation levels regulated by an isopropyl β-d-1-thiogalactopyranoside-inducible lac promoter. (B) Diverse read-throughs of RITs at different transcription initiation levels. (C) Plasmid design to measure the decay rate of messenger RNAs (mRNAs) with different 3′-untranslated regions (UTRs). (D) The fluorescence intensity of green fluorescent protein (GFP) differed depending on the 3′-UTR. Error bars indicate the standard deviation of two biological replicates. (E) Exponential decay of mRNA from pGFP. Error bars indicate the standard deviation of two biological replicates, each composed of technically triplicate qRT-PCR reactions. T1/2 indicates the mRNA half-life. (F) Fluorescence intensity was directly correlated with the half-life of mRNA encoding GFP. (G) Computational modeling of fluorescence protein expression showed a reliable prediction of the experimental measurements. Circles indicate experimental values from pGFP_empty and pGFP_rrnBT. Shades are ranges of the possible fluorescence intensity set by the standard error of the measured half-lives.