Part:BBa_K4174001:Experience

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William and Mary iGEM 2022

To test the effectiveness of our mRFP1 construct (BBa_K4174001), our team transformed the original MIT iGEM 2006 construct (BBa_J45995), our mRFP1 construct, and our sfGFP construct (BBa_K4174001) into E. coli NEB5α cells and grew the various transformants in a plate reader. They were grown at 37°C. For red fluorescence measurements, we used an excitation value of 584 nm and an emission value of 610 nm. For green fluorescence measurements, we used an excitation value of 485 nm and an emission value of 528 nm. The values for red fluorescence are reported below. For information about green fluorescence measurements, see parts page BBa_K4174002.

Based on the graph above, the bacterial cells engineered with our mRFP1 construct appear to have entered stationary phase around 16 hours. As seen in the graph, our mRFP1 construct produces more red fluorescence than the original circuit. The other measurements taken are for our sfGFP construct and untransformed E. coli NEB5α cells, both of which serve as negative controls for red fluorescence.

As seen in the image above, qualitative results reveal that our mRFP1 construct produces more red fluorescence than the original construct. Here, our mRFP1 construct is on the far left, and is visibly more red that the original GFP construct.

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