Difference between revisions of "Part:BBa K2924016"
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===Characterization===
 
===Characterization===
 
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The promoter was tested for the sensitivity to butyric acid in the culture medium by combining the promoter to an eYFP (<a href=”https://parts.igem.org/Part:BBa_E0030”>BBa_E0030</a>) <sup>2</sup> as a reporter gene in the composite part <a href=”https://parts.igem.org/Part:BBa_K2924017”>BBa_K2924017</a>. The concentrations of butyric acid were from 0.5 mM to 20 mM.
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The promoter was tested for the sensitivity to butyric acid in the culture medium by combining the promoter to an eYFP (<a href=”https://parts.igem.org/Part:BBa_E0030”>BBa_E0030</a>) <sup>2</sup> as a reporter gene in the composite part <a href=”https://parts.igem.org/Part:BBa_K2924017”>BBa_K2924017</a>. The concentrations of butyric acid were from 0.5 mM to 20 mM.</html>
  
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[[File:Flic_c4.png|500px|thumb|left|<i><b>Fig.2:</b> Response of P<sub>fliC</sub>+eYFP (red) to different chain lengths of fatty acids compared to an empty vector control (black). The fluorescence was measured at an excitation wavelength from 497 nm and an emission wavelength from 540 nm.</i>]]
  
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The experiment showed that the fluorescence doesn't grow with higher concentrations of butyric acid. Surprisingly the fluorescence from the empty vector control rises with higher concentrations while the P<sub>fliC</sub> shows a falling tendency.
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[[File:FadR_mechanism.png|600px|thumb|left|<i><b>Fig.1:</b> Regulation of the fatty acid metabolism by the transcriptional factor FadR. FadR recognizes its cognate binding site (white), thereby repressing transcription. Upon binding of an Acyl-CoA to FadR, the promoter region is freed, enabling gene expression.</i>]]
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[[File:Flic_c4.png|300px|thumb|right|<i><b>Fig.2:</b> Response of P<sub>fliC</sub>+eYFP (red) to different chain lengths of fatty acids compared to an empty vector control (black). The fluorescence was measured at an excitation wavelength from 497 nm and an emission wavelength from 540 nm.</i>]]
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Revision as of 19:04, 19 October 2019


Promoter fliC from the Escherichia coli genome

Short-chain fatty acid sensitive promoter FliC

Usage and Biology

The promoter fliC was published as a sensitive promoter for short-chain fatty acids, especially for butyrate (C4:0). This promoter was isolated from the Escherichia coli wild type genome. In the wild type the short-chain fatty acids have an impact on the flagellar expression. The PfliC is repressed by leucine-responsive regulatory protein (Lrp). Butyrate can enhance the expression of the flagellar expression like leucine which is a ligand of Lrp. Difference between thus enhancers is that the promoter fliC is only sensitive for the butyrate and not for the leucine 1>/sup>

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Characterization

The promoter was tested for the sensitivity to butyric acid in the culture medium by combining the promoter to an eYFP (BBa_E0030) 2 as a reporter gene in the composite part BBa_K2924017. The concentrations of butyric acid were from 0.5 mM to 20 mM.

Fig.2: Response of PfliC+eYFP (red) to different chain lengths of fatty acids compared to an empty vector control (black). The fluorescence was measured at an excitation wavelength from 497 nm and an emission wavelength from 540 nm.

The experiment showed that the fluorescence doesn't grow with higher concentrations of butyric acid. Surprisingly the fluorescence from the empty vector control rises with higher concentrations while the PfliC shows a falling tendency.