Part:BBa_K2100005:Experience

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Applications of BBa_K2100005

The MIT iGEM team used the hEF1a promoter to constitutively express fluorescent proteins as transfection markers in our experiments in HEK293, tHESC, MCF-7, and ISH cells. The amount of fluorescent units observed is an indication of the number of plasmids a particular cell uptakes. The graph below shows two single color controls from one of our experiments. We transfected one well of HEK293 cells with hEF1a-mKate and another with hEF1a-eYFP. The cells that uptook the hEF1a-mKate plasmids showed an increase in only the red fluorescent output, while the cells that uptook the hEF1a-eYFP plasmids showed a clear increase in only the yellow fluorescent output.

Here is an image of HEK293 cells transfected with the eYFP florescent gene cascaded with a hEF1a constituitive promoter. We often used yellow florescence as either a reporter for how our promoter responded to induction in cells or as transfection markers for experiments.

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