Part:BBa_K1045017:Experience

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Applications of BBa_K1045017

To characterize the DarR reporter system, E. coli was transformed either with part BBa_K1045017 or with part 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 seemed to act as a strong repressor in E. coli even in the absence of cyclic do-AMP.

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

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

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