Team:LCG-UNAM-Mexico/Experiments/Design
From 2008.igem.org
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<p><span class="calHeader"><a name="Devices"></a>System</span></p> | <p><span class="calHeader"><a name="Devices"></a>System</span></p> | ||
- | <p align="justify"> First of all, we needed a system that could <span dir="ltr" id=":1s">cause a change in its medium | + | <p align="justify"> First of all, we needed a system that could <span dir="ltr" id=":1s">cause a change in its medium conductivity</span>. An extrusion pump seemed to be the best way to achieve this. Once this was devised, we needed a way to regulate the system. <span dir="ltr" id=":1s">We decided to use a negative regulator because it's the only way to transcriptionally regulate the expression of a gene in a definitive way.</span></p> |
<p align="justify"><br> | <p align="justify"><br> | ||
- | We had to be able to restart our system, so we could add a signal at anytime. This could be accomplished with an induction signal that | + | We had to be able to restart our system, so we could add a signal at anytime. This could be accomplished with an induction signal that disappears rapidly after its involvement. The need of a link between the inductor signal and the repressor, lead us to include a little regulation cascade. This cascade allows us to add new steps which might increase our system’s complexity.<br> |
</p> | </p> | ||
<p>The components selected to fulfill the system requirements are enlisted in the next table:</p> | <p>The components selected to fulfill the system requirements are enlisted in the next table:</p> | ||
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<p align="left"><span class="style3">Device <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K119009">BBa_K119009</a>: <em>The extrusion pump.</em></span></p> | <p align="left"><span class="style3">Device <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K119009">BBa_K119009</a>: <em>The extrusion pump.</em></span></p> | ||
<p align="justify"><br> | <p align="justify"><br> | ||
- | The propose of this device is to manipulate the transcription of rcnA by an inhibitory signal while maintaining the natural regulation of rcnA through RcnR. To achieve this the device contains a CI dependent promoter, RcnR binding site and the RcnA extrusion pump inserted in the vector <a href="https://static.igem.org/mediawiki/2008/9/94/ | + | The propose of this device is to manipulate the transcription of rcnA by an inhibitory signal while maintaining the natural regulation of rcnA through RcnR. To achieve this the device contains a CI dependent promoter, RcnR binding site and the RcnA extrusion pump inserted in the vector <a href="https://static.igem.org/mediawiki/2008/9/94/PBB1MCS-5.PNG">pBBR1MCS-5.</a></p> |
<p align="justify" class="style3"> </p> | <p align="justify" class="style3"> </p> | ||
<p align="justify" class="style3">Devices <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K119010">BBa_K119010</a>/<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K119010">BBa_K119011</a>: <em>The regulatory device</em></p> | <p align="justify" class="style3">Devices <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K119010">BBa_K119010</a>/<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K119010">BBa_K119011</a>: <em>The regulatory device</em></p> | ||
- | <p align="justify" class="bodyText">In order to control the RcnA activity this device includes the gene encoding LuxR under the regulation TetR constitutive promoter followed by cI, which will repress RcnA in the prescence of AHL:LuxR. The last component of the device is the gene encoding AiiA. In <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K119010">BBa_K119010</a> lacZ promoter is upstream of AiiA, while <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K119010">BBa_K119011</a> carries a mutated version of it. The plasmid carrying this device will be PRK415.</p> | + | <p align="justify" class="bodyText">In order to control the RcnA activity this device includes the gene encoding LuxR under the regulation TetR constitutive promoter followed by cI, which will repress RcnA in the prescence of AHL:LuxR. The last component of the device is the gene encoding AiiA. In <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K119010">BBa_K119010</a> lacZ promoter is upstream of AiiA, while <a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K119010">BBa_K119011</a> carries a mutated version of it. The plasmid carrying this device will be <a href="https://static.igem.org/mediawiki/2008/5/5e/PRK415.png">PRK415</a>.</p> |
<p align="left"> </p> | <p align="left"> </p> | ||
<p align="left"> </p> | <p align="left"> </p> |
Revision as of 14:40, 29 October 2008
LCG-UNAM-Mexico | ||||||||||||||||
iGEM 2008 TEAM | ||||||||||||||||
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System First of all, we needed a system that could cause a change in its medium conductivity. An extrusion pump seemed to be the best way to achieve this. Once this was devised, we needed a way to regulate the system. We decided to use a negative regulator because it's the only way to transcriptionally regulate the expression of a gene in a definitive way.
The components selected to fulfill the system requirements are enlisted in the next table:
Devices
Device BBa_K119009: The extrusion pump.
Devices BBa_K119010/BBa_K119011: The regulatory device In order to control the RcnA activity this device includes the gene encoding LuxR under the regulation TetR constitutive promoter followed by cI, which will repress RcnA in the prescence of AHL:LuxR. The last component of the device is the gene encoding AiiA. In BBa_K119010 lacZ promoter is upstream of AiiA, while BBa_K119011 carries a mutated version of it. The plasmid carrying this device will be PRK415.
References
2.-Rodrigue A. Et al.”Identification of rcnA (yohM), a Nickel and Cobalt Resistance Gene in Esherichia coli” 2005. 3.-Kovach et al.,”pBBR1MCS: a broad-host-range cloning vector”.1994 4.- 5.-link a chiba, ahorita lo pongo 6.-Whiteheada N.A., Barnada A.M.L., Slaterra H..”Quorum-sensing in Gram-negative bacteria”2001. 16.-N.T. Keen, S. Tamaki, D. Kobayashi, and D. Trollinger. 1998.
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