Part:BBa_K1045017:Experience

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Applications of BBa_K1045017

To characterize the DarR reporter system, E. coli was transformed either with BBa_K1045017 or BBa_K1045013 as a control.

In BBa_K1045013, gfp is placed downstream of a strong promoter and the DarR operator. This vector does not encode for DarR. The strong fluorescence of the cells transformed with BBa_K1045013 (Fig. 1) might indicate that GFP was expressed.
However, when transformed with BBa_K1045017 (Fig. 2), the cells showed almost no fluorescence. In contrast to BBa_K1045013, BBa_K1045017 encodes for DarR. The low fluorescence might hint that DarR was expressed and active as a repressor down-regulating gfp transcription. Hence, DarR seems to act as a strong repressor in E. coli even in the absence of cyclic di-AMP.

Fig. 1. - DarR
E. coli transformed with a plasmid encoding BBa_K1045013 shows a strong green fluorescence under the fluorescence microscope

Fig. 2. + DarR
E. coli transformed with a plasmid harboring the DarR reporter system shows barely fluorescence.

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