Difference between revisions of "Part:BBa K861060:Experience"
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<h4>Result</h4>
 
<h4>Result</h4>
 
<p align="justify"> Normalized using Fluorescence/0D600</p>
 
<p align="justify"> Normalized using Fluorescence/0D600</p>
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<p align="justify"> Blue: Constitutive promoter J23110</p>
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<p align="justify"> Green: PfadR</p>
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<p align="justify"> Red: PfadR</p>
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<p align="justify"> Glucose Concentration gradient: 1, 10 mM</p>
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<p align="justify"> Glucose Concentration gradient: 0.5, 1, 5, 10 mM</p>
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<p align="justify"> Fatty acid Concentration gradient: 0.025, 0.05, 0.1, 0.5,1.5 mM</p>
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<p align="justify"> Fatty acid Concentration gradient: 0.025, 0.05, 0.1, 0.25, 0.5, 1, 1.5 mM</p>
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<p><img src="https://static.igem.org/mediawiki/parts/0/00/PfadR.png" width="500" height="350" hspace="2" vspace="1" border="2" align="top" /></p>
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<p><img src="https://static.igem.org/mediawiki/parts/6/6f/Pfadr.png" width="500" height="350" hspace="2" vspace="1" border="2" align="top" /></p>
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<p><i> PfadR and BBa_J23110 promoter strength at different glucose and fatty acids concentration</i></p>
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<p><i> PfadR promoter strength at different glucose and fatty acids concentration</i></p>
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<p align="justify"> As can be shown from the results, the promoter shows about 3 times induction from glucose to 1.5umol/L fatty acids medium and the fluorescence of PfadR is about one sixth of <a href="https://parts.igem.org/wiki/index.php?title=Part:BBa_J23110">BBa_J23110</a>. This may be result from the tandem FadR binding site has made it more difficult for Polymerase to start gene transcription. Also, in medium that use glucose as sole carbon source, PfadR seems to be leaky. However, since our bacteria is wild type <i>E.coli</i>, Fab genes was not mutated, meaning that bacteria can synthesis fatty acids. Therefore, there may be a basal level fatty acids concentration inside the cell, making the transcription not being totally repressed. </p>
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<p align="justify"> As can be shown from the results, the promoter shows about 3 times induction from glucose to 1.5umol/L fatty acids medium and the fluorescence of PfadR. Also, in medium that use glucose as sole carbon source, PfadR seems to be leaky. However, since our bacteria is wild type <i>E.coli</i>, Fab genes was not mutated, meaning that bacteria can synthesis fatty acids. Therefore, there may be a basal level fatty acids concentration inside the cell, making the transcription not being totally repressed. </p>
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<p align="justify"> It should also be noticed that from fatty acids concentration 0.025umol/L to 1.5umol/L, the induction is not very obvious. F0.25, 0.5 and 1 seemed to have similar fluorescence strength. The promoter is not sensitive enough.  For more details, please visit our <a href="http://2012.igem.org/Team:WHU-China/Project?catalog=2">wiki</a>.</p>
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<p align="justify">For more details, please visit our <a href="http://2012.igem.org/Team:WHU-China/Project?catalog=2">wiki</a>.</p>
  
 
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Revision as of 03:55, 30 October 2012

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Applications of BBa_K861060

Result

Normalized using Fluorescence/0D600

Green: PfadR

Glucose Concentration gradient: 1, 10 mM

Fatty acid Concentration gradient: 0.025, 0.05, 0.1, 0.5,1.5 mM

PfadR promoter strength at different glucose and fatty acids concentration

As can be shown from the results, the promoter shows about 3 times induction from glucose to 1.5umol/L fatty acids medium and the fluorescence of PfadR. Also, in medium that use glucose as sole carbon source, PfadR seems to be leaky. However, since our bacteria is wild type E.coli, Fab genes was not mutated, meaning that bacteria can synthesis fatty acids. Therefore, there may be a basal level fatty acids concentration inside the cell, making the transcription not being totally repressed.

For more details, please visit our wiki.

User Reviews

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