Part:BBa_K3788020:Experience

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Applications of BBa_K3788020

Method

In order to study the previously reported latency period between induction of the caa-cal operon system, ColicinA production, and CaL lysis protein production, we fused the RFP coding sequence (BBa_E1010) with the Modified lysis device part (BBa_K3788015) (composite part BBa_K3788020). Induction of the SOS response was induced by mitomycin C treatments and fluorescence was recorded as a function of dose using a microplate reader.

Results

Cells carrying the BBa_K3788020 construct were cultured in the presence of various concentrations of mitomycin C, ranging from 0 to 800 ng.ml-1. The OD (600 nm) and the amount of red fluorescence were recorded over time.

We observed a strong dose-dependent synthesis of our reporter protein during treatment with mitomycin C. The signal was detected after 330 min to 420 min of induction, for the highest dose and the lowest dose, respectively.

This contrasts with what we observed when GFP was under the control of the LexA-dependent promoter (BBa_K3780817) where we could observe the accumulation of green fluorescence as early as 40 minutes after induction.

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