Part:BBa_K1045011:Experience

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Applications of BBa_K1045011

We used this part in our DarR reporter system BBa_K1045017. BBa_K1045011 was functional as our characterization experiments of BBa_K1045017 suggested. In this system, gfp is placed under the control of the DarR operator BBa_K1045000. We saw that GFP expression was repressed when E. coli harbored a construct carrying DarR under the control of the promoter in BBa_K1045011. For more information, see main page of BBa_K1045017 and our entry on the experience page of BBa_K1045017:

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 indicated that GFP was expressed. However, when transformed with BBa_K1045017 (Fig. 1), the cells showed almost no fluorescence. In contrast to BBa_K1045013, BBa_K1045017 encodes for DarR. The low fluorescence suggested 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.: Top: E. coli transformed with a plasmid encoding BBa_K1045013 shows a strong green fluorescence under the fluorescence microscope. Bottom: E. coli transformed with a plasmid harboring the DarR reporter system barely shows fluorescence.

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