Difference between revisions of "Part:BBa K1045011:Experience"

Revision as of 17:38, 12 October 2013

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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.

User Reviews

UNIQccba03404dbab36b-partinfo-00000000-QINU UNIQccba03404dbab36b-partinfo-00000001-QINU

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 We used this part in our DarR reporter system [[Part:BBa_K1045017|BBa_K1045017]]. [[Part:BBa_K1045011|BBa_K1045011]] was functional as our characterization experiments of [[Part:BBa_K1045017|BBa_K1045017]] suggested. In this system, ''gfp'' is placed under the control of the DarR operator [[Part:BBa_K1045000|BBa_K1045000]]. We saw that GFP expression was repressed when ''E. coli'' harbored a construct carrying DarR under the control of the promoter in [[Part:BBa_K1045011|BBa_K1045011]]. For more information, see main page of [[Part:BBa_K1045017|BBa_K1045017]] and our entry on the experience page of BBa_K1045017:
+In [[Part:BBa_K1045013|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 [[Part:BBa_K1045013|BBa_K1045013]] indicated that GFP was expressed. However, when transformed with [[Part:BBa_K1045017|BBa_K1045017]] ('''Fig. 1'''), the cells showed almost no fluorescence. In contrast to [[Part:BBa_K1045013|BBa_K1045013]], [[Part:BBa_K1045017|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.
 [[File:-DarR.jpg|420px|thumb|'''Fig. 1.''': ''Top'': ''E. coli'' transformed with a plasmid encoding [[Part:BBa_K1045013|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. [[File:+DarR.jpg|420px]]|center]]