Team:MIT k415031

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<center><a href="http://2010.igem.org/wiki/images/5/5a/Regulatory_yo.png" class="thickbox" title="Regulatory promoter pLux/cI-OR"><img src="http://2010.igem.org/wiki/images/5/5a/Regulatory_yo.png"></a><br><br>
<center><a href="http://2010.igem.org/wiki/images/5/5a/Regulatory_yo.png" class="thickbox" title="Regulatory promoter pLux/cI-OR"><img src="http://2010.igem.org/wiki/images/5/5a/Regulatory_yo.png"></a><br><br>
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<b class="bolded"> K415031 is less leaky than R0065 </b><hr>
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<b class="bolded"> K415031 is less leaky than R0065 </b>
<a href="http://2010.igem.org/wiki/images/a/af/Leakiness.png" class="thickbox"><img src="http://2010.igem.org/wiki/images/a/af/Leakiness.png"></a><br>
<a href="http://2010.igem.org/wiki/images/a/af/Leakiness.png" class="thickbox"><img src="http://2010.igem.org/wiki/images/a/af/Leakiness.png"></a><br>

Latest revision as of 03:46, 27 October 2010

K415031

From the 2010 MIT iGEM Team.See the Parts Registry Page!! →

Improved PLux/cI-OR Promoter
This part is a promoter that was designed as a correction to the anomalous BBa_R0065 BioBrick. This part consists of a Lux Box binding site for the LuxR-C6HSL Complex; in addition, it contains two operator sites for the cI protein appropriated from the Lambda bacteriophage. Because basal expression without the LuxR-C6HSL complex present is expected to be minimal, and because the cI operator sites are positioned so as to block the critical -10 transcription initiation site on the promoter, if cI is high, the promoter is off; if AHL is low and cI is absent, the promoter is leaky; if AHL is high and cI is absent, the promoter is on.

Slightly higher binding than K415032.




K415031 is less leaky than R0065
The above images are FACS scatter plots of co-transformations of either pTSMa (BBa_K415300) and K415069 or pTSMa and BBa_K415023. The x-axis represents the amount of green fluorescence, and the y-axis represents the amount of red fluorescence. The parts K415069 and K415023 differ only in a single promoter sequence: K415069 has the K415031 promoter in place of a single R0065 promoter in the K415023 sequence. In this way, the more "leaky" the promoter, the more red fluorescing cells.
As you can see in the images, fewer cells with pTSMa+K415069 are leaky: 3% of total cells with pTSMa+K415069 fluoresce red, and 15% of total cells with pTSMa+K415023 fluoresce red.


← K415301K415032 →