Part:BBa_K2235003:Experience

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Applications of BBa_K2235003

At first, we have measured the cell viability of TOP10 in different concentrations of cumate over time. Our aim was to determine whether cumate is cytotoxic or not, before we measure the fluorescence.

The growth curve clearly demonstrates that cumate does not affect the cellular viability and proliferation of our strain.

Next, we employed the same concentrations and measured the fluorescence over time. We also used a negative control (transformed bacteria which is not expressing BFP).

The fluorescence intensity is higher in all samples compared to the negative control. Therefore, the expression of BFP is not in a dose dependent manner since fluorescence can be detected even in the untreated sample. Normally, we would expect to see fluorescence only in the treated samples since the cumate binds to the repressor enabling the transcription of the downstream gene. However, we suggest that the cymR is mutated at a specific site based on the sequencing results; thus, the expressed protein is unable to bind to the T5 operon in the first place and suppress the expression of BFP.

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