Difference between revisions of "Part:BBa K1045017:Experience"

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===Applications of BBa_K1045017===
 
===Applications of BBa_K1045017===
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To characterize the DarR reporter system, ''E. coli'' was transformed either with part BBa_K1045017 or with part BBa_K1045013 as a control.
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To characterize the DarR reporter system, ''E. coli'' was transformed either with [[Part:BBa_K1045017|BBa_K1045017]] or [[Part:BBa_K1045013|BBa_K1045013]] as a control.
  
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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.<br />
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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]] (Fig. 1) might indicate that GFP was expressed.<br />
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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.<br />
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However, when transformed with [[Part:BBa_K1045017|BBa_K1045017]] (Fig. 2), 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 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.<br />
  
  
  
 
[[File:-DarR.jpg|420px|thumb|left|'''Fig. 1.''' - DarR <br />
 
[[File:-DarR.jpg|420px|thumb|left|'''Fig. 1.''' - DarR <br />
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''E. coli'' transformed with a plasmid encoding part BBa_K1045013 show a strong green fluorescence under the fluorescence microscope]]
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''E. coli'' transformed with a plasmid encoding [[Part:BBa_K1045013|BBa_K1045013]] show a strong green fluorescence under the fluorescence microscope]]
 
[[File:+DarR.jpg|420px|thumb|right|'''Fig. 2.''' + DarR <br />
 
[[File:+DarR.jpg|420px|thumb|right|'''Fig. 2.''' + DarR <br />
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''E. coli'' transformed with a plasmid harboring the DarR reporter system barely fluoresce.]]
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''E. coli'' transformed with a plasmid harboring the DarR reporter system barely fluorescence.]]
  
 
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Revision as of 16:12, 25 September 2013


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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 show a strong green fluorescence under the fluorescence microscope
Fig. 2. + DarR
E. coli transformed with a plasmid harboring the DarR reporter system barely fluorescence.





























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