Difference between revisions of "Part:BBa K1045000:Experience"

Revision as of 13:20, 16 October 2013

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Applications of BBa_K1045000

We used this Biobrick in our DarR reporter system (BBa_K1045017). When characterizing this system in E. coli, we noticed that the DarR operator sequence as it is in BBa_K1045000 seems to be strongly bound by DarR (BBa_K1045001) even in the absence of c-di-AMP.

The experimental setup used for characterization involved two different E. coli strains: 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 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

UNIQb2fc7ea54c452eaf-partinfo-00000000-QINU UNIQb2fc7ea54c452eaf-partinfo-00000001-QINU

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 ===Applications of BBa_K1045000===
-We used this Biobrick in our DarR reporter system ([[Part:BBa_K1045017|BBa_K1045017]]). When characterizing this system in ''E. coli'', we noticed that the DarR operator sequence as it is in '''BBa_K1045000''' seems to be strongly bound by DarR ([[Part:BBa_K1045001|BBa_K1045001]]) even in the absence of c-di-AMP. For experimental data, read our entry on [https://parts.igem.org/Part:BBa_K1045017:Experience].
+We used this Biobrick in our DarR reporter system ([[Part:BBa_K1045017|BBa_K1045017]]). When characterizing this system in ''E. coli'', we noticed that the DarR operator sequence as it is in '''BBa_K1045000''' seems to be strongly bound by DarR ([[Part:BBa_K1045001|BBa_K1045001]]) even in the absence of c-di-AMP.
+The experimental setup used for characterization involved two different ''E. coli'' strains: ''E. coli'' was transformed either with [[Part:BBa_K1045017|BBa_K1045017]] or [[Part:BBa_K1045013|BBa_K1045013]] as a control.
+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]]
 ===User Reviews===