Gene expression can be regulated in a variety of ways, including by RNA thermometers (RNATs). RNATs are elements that act in a temperature-dependent fashion by changing their structure. They primarily affect translation, forming folded structures that block the ribosome binding site at lower temperatures. Thermometer structures include hairpins, stems, and loops which melt open at higher temperatures and allow translation. Here, reporter assays with green fluorescent protein (GFP) are investigated to study the structure-function relationship of RNATs. The thermometer melting directs the production of GFP, leading to fluorescence of <i>E. coli</i> cells. We present work looking at the behavioral response of synthetic thermometers in real time by directly monitoring fluorescence intensity and cell density of bacterial cultures. Two different fluorescent plate readers were used to examine the robustness of our reporter assay. Our results highlight overall trends of reporter gene expression with response to temperature. We found that model thermometers demonstrate greater fold increase in fluorescence between 30 and 37°C growths than a control lacking the RNAT. We are continuing to study the differences of equipment, various growth conditions, and the effect maturation rate of GFP has on real-time analysis.

Studying GFP production in <i>E. coli</i> for a more direct correlation to RNA thermometer behavior

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Studying GFP production in E. coli for a more direct correlation to RNA thermometer behavior