Team:Grenoble/Projet/Intro

From 2011.igem.org

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Ne laissez pas le carré trop longtemps si vous n'éditez pas, chaque fois reprenez ce qui est sur internet plutôt que ce que vous avez sur votre PC
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<div class="body">
<div class="body">
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<h2>The Project, Mercuro-Coli</h1>
 
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<div class="blocbackground">
 
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<div class="blocbackground">
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<h1>The Project, Mercuro-Coli</h1>
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<h2>Introduction</h2>
<h2>Introduction</h2>
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<a href="https://static.igem.org/mediawiki/2011/5/58/Dispositif.png"><img src="https://static.igem.org/mediawiki/2011/5/58/Dispositif.png" width=600px/></a>
<a href="https://static.igem.org/mediawiki/2011/5/58/Dispositif.png"><img src="https://static.igem.org/mediawiki/2011/5/58/Dispositif.png" width=600px/></a>
</center>
</center>
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<div class="legend">
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Easy to use in three steps!
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</div>
<ul>
<ul>
<li>
<li>
-
The device consists of a simple packaged plate.
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Unpack the plate.
</li>
</li>
<li>
<li>
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The polluted water sample is added to the unpacked plate.
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Deposit the polluted water sample.
</li>
</li>
<li>
<li>
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The appearance of a red line on the plate signifies the presence of mercury in the sample.
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If mercury is present, a red stripe appear, its position indicates the amount of pollutant.
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</li>
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<li>
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The red line position on the plate indicates the pollutant quantity.
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</li>
</li>
</ul>
</ul>
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<p>
<p>
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At the beginning of the project, we defined a certain number of specifications for the device, which we describe below.
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At the beginning of the project, we defined a certain number of specifications for the device:
</p>
</p>
<ul>
<ul>
<li>
<li>
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<strong>Operating living cell: E. Coli: BW 25 113</strong>
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<strong>Use a single bacterial strain: <i>Escherichia coli</i> BW 25 113, containing the whole genetic network</strong>
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</li>
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<li>
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<strong>Only one type of bacteria containing the whole genetic circuit.</strong>
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</li>
</li>
<li>
<li>
<strong>Form a biofilm on the plate with <i>E. coli</i> bacteria containing the designed genetic circuit.</strong><br/>
<strong>Form a biofilm on the plate with <i>E. coli</i> bacteria containing the designed genetic circuit.</strong><br/>
-
This point was revised according to modeling results. A device with channels containing bacteria instead of a biofilm has been chosen.  
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This point was revised according to <a href="https://2011.igem.org/Team:Grenoble/Projet/Results/Quorum#Simulation" title="Optimization of the device thanks to modelling">modeling results.</a> A device with channels containing bacteria instead of a biofilm has been chosen.  
</li>
</li>
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</ul>
</ul>
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<center>
<a href="https://static.igem.org/mediawiki/2011/c/c5/1_type_bacteria.png"><img align="middle" src="https://static.igem.org/mediawiki/2011/c/c5/1_type_bacteria.png" width=600px/></a>
<a href="https://static.igem.org/mediawiki/2011/c/c5/1_type_bacteria.png"><img align="middle" src="https://static.igem.org/mediawiki/2011/c/c5/1_type_bacteria.png" width=600px/></a>
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</center>
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<div class="legend">
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Plate covered by the bacteria.
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</div>
<ul>
<ul>
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<li><strong>Comparative measure:</strong></li>
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<li><strong>Build a comparative measurement system:</strong></li>
</ul>
</ul>
<table class="nobordure">
<table class="nobordure">
<tr>
<tr>
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<td><a href="https://static.igem.org/mediawiki/2011/7/7d/Comparaison.png"><img src="https://static.igem.org/mediawiki/2011/7/7d/Comparaison.png"  height="300px"></a></td>
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<td><a href="https://static.igem.org/mediawiki/2011/0/07/Plaque_mer_iptg.png"><img src="https://static.igem.org/mediawiki/2011/0/07/Plaque_mer_iptg.png"  height="350px"></a></td>
<td>
<td>
<p>
<p>
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The comparative measure will be made by comparing mercury concentration to IPTG concentration. IPTG concentration will is the reference.
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The principle of the measurement is based on the comparison between an unknown mercury concentration and a known IPTG concentration. In practice, the quantification scale is made by the prior application of a concentration gradient of IPTG on the plate.
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</p>
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-
<p>
+
-
So, an IPTG gradient concentration will be made beforehand in the plate constituting our scale of quantification.
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</p>
</p>
</td>
</td>
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<p>
<p>
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By adding uniformly the polluted sample, two different bacteria behaviors will come up depending on the predominant concentration:
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Two different bacterial behaviors arise from the uniform addition of polluted sample on the plate. They depend on the predominant concentration:
</p>
</p>
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<ul>
<ul>
<li>
<li>
-
On the left side, where the mercury concentration is prevailing, bacteria with a <strong>sending</strong> behavior will appear. They have the ability to release in the external medium Quorum Sensing molecules, AHL, thanks to the expression of the CinI protein.
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On the left side, where the mercury concentration is prevailing, bacteria will behave as <strong>senders</strong>. They have the ability to release quorum sensing molecules (AHL) in the external medium, thanks to the expression of the CinI protein.
</li>
</li>
<li>
<li>
-
On the right, where the IPTG concentration is dominant, bacteria with a <strong>receiving</strong> behavior will come up. They express CinR protein, a cytoplasmic receptor for Quorum Sensing.
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On the right, where the IPTG concentration is dominant, bacteria will behave as <strong>receivers</strong>. They express CinR protein, a cytoplasmic receptor for quorum sensing.
</li>
</li>
</ul>
</ul>
</td>
</td>
<td>
<td>
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<a href="https://static.igem.org/mediawiki/2011/8/81/Receiving_secr_bacteria.svg.png"><img src="https://static.igem.org/mediawiki/2011/8/81/Receiving_secr_bacteria.svg.png"  width="350px"></a>
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<center>
 +
<a href="https://2011.igem.org/Team:Grenoble/Projet/Design/quorum" title="Click for more details on the relation between the two bahaviours and the quorum sensing"><img src="https://static.igem.org/mediawiki/2011/0/00/Receiving_secr_bacteria.png"  width="450px"></a>
 +
</center>
</td>
</td>
</tr>
</tr>
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<td>
<td>
<p>
<p>
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So, we will have the emergence of two distinctive areas separated by one boundary that can appear closer to one side or the other depending on the quantity of mercury. For example, if the sample contains less mercury the boundary will appear closer to the left side.
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In summary, two areas with distinctive bacterial behavior appear on the plate. They are separated by one boundary that approaches one side of the plate or the other, depending on the mercury level. For example, in the case of a sample with a low mercury concentration, the boundary will appear closer to the left side.
</p>
</p>
</td>
</td>
<td>
<td>
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<a href="https://static.igem.org/mediawiki/2011/d/d8/Border_movement.png"><img src="https://static.igem.org/mediawiki/2011/d/d8/Border_movement.png"  width="350px"></a>
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<a href="https://static.igem.org/mediawiki/2011/d/d8/Border_movement.png"><img src="https://static.igem.org/mediawiki/2011/d/d8/Border_movement.png"  width="450px"></a>
</td>
</td>
</tr>
</tr>
</table>
</table>
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<p>So, the location of that boundary represents our point of interest that we will try to visualize.</p>
 
 +
<p>
 +
The key point is therefore the localization of the boundary, which we need to visualize
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</p>
<ul>
<ul>
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</ul>
</ul>
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<p>The two different behaviors were chosen in purpose.</p>
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<center>
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<table class="nobordure">
<table class="nobordure">
<tr>
<tr>
<td>
<td>
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<a href="https://static.igem.org/mediawiki/2011/0/02/Coloration.png"><img src="https://static.igem.org/mediawiki/2011/0/02/Coloration.png"  width="300px"></a>
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<a href="https://2011.igem.org/Team:Grenoble/Projet/Design/quorum#coloration" title="click for more details on the appearance of the red stripe"><img src="https://static.igem.org/mediawiki/2011/0/02/Coloration.png"  width="400px"></a>
</td>
</td>
<td>
<td>
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<a href="https://static.igem.org/mediawiki/2011/c/c2/Complex.png"><img src="https://static.igem.org/mediawiki/2011/c/c2/Complex.png"  width="260px"></a>
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<a href="https://2011.igem.org/Team:Grenoble/Projet/Design/quorum#coloration" title="click for more details on the appearance of the red stripe"><img src="https://static.igem.org/mediawiki/2011/c/c2/Complex.png"  width="340px"></a>
</td>
</td>
</tr>
</tr>
</table>
</table>
 +
</center>
<p>
<p>
-
At the boundary,Quorum Sensing molecule, AHL, released by sending bacteria will be received by the front of receiving bacteria. The complex formed by AHL molecule and CinR protein will then induce the coloration of the front receiving bacteria.
+
At the boundary,Quorum Sensing molecule, AHL, released by sending bacteria will be uptaken by the front of receiving bacteria. The complex formed by AHL molecules and CinR proteins will then induce the coloration of the front receiving bacteria.
-
</p>
+
-
<p>
+
-
Let's go further to understand how such evolution of this system is possible with only one strain ?
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</p>
</p>
 +
 +
<center>
 +
<a href="https://2011.igem.org/Team:Grenoble/Projet/Design" title="Click here"><img src="https://static.igem.org/mediawiki/2011/1/1b/Bouton_general_biologie.png"/></a>
 +
<div class="legend">
 +
Let's go further to understand how such evolution of this system is possible with only one strain ?
 +
</div>
 +
</center>
</div>
</div>

Latest revision as of 23:03, 28 October 2011

Grenoble 2011, Mercuro-Coli iGEM


The Project, Mercuro-Coli

Introduction

With industrial growth, wastes are accumulating and the presence of pollutant and toxic compounds is becoming an international concern.

In this context, we present Mercuro-Coli, an easy-to-use biosensor for the in situ detection and quantification of mercury into polluted water. Intended to fieldwork applications, the device should be very easy to handle:

Easy to use in three steps!
  • Unpack the plate.
  • Deposit the polluted water sample.
  • If mercury is present, a red stripe appear, its position indicates the amount of pollutant.

Specifications

At the beginning of the project, we defined a certain number of specifications for the device:

  • Use a single bacterial strain: Escherichia coli BW 25 113, containing the whole genetic network
  • Form a biofilm on the plate with E. coli bacteria containing the designed genetic circuit.
    This point was revised according to modeling results. A device with channels containing bacteria instead of a biofilm has been chosen.
Plate covered by the bacteria.
  • Build a comparative measurement system:

The principle of the measurement is based on the comparison between an unknown mercury concentration and a known IPTG concentration. In practice, the quantification scale is made by the prior application of a concentration gradient of IPTG on the plate.

Two different bacterial behaviors arise from the uniform addition of polluted sample on the plate. They depend on the predominant concentration:

  • On the left side, where the mercury concentration is prevailing, bacteria will behave as senders. They have the ability to release quorum sensing molecules (AHL) in the external medium, thanks to the expression of the CinI protein.
  • On the right, where the IPTG concentration is dominant, bacteria will behave as receivers. They express CinR protein, a cytoplasmic receptor for quorum sensing.

In summary, two areas with distinctive bacterial behavior appear on the plate. They are separated by one boundary that approaches one side of the plate or the other, depending on the mercury level. For example, in the case of a sample with a low mercury concentration, the boundary will appear closer to the left side.

The key point is therefore the localization of the boundary, which we need to visualize

  • A visual result:

At the boundary,Quorum Sensing molecule, AHL, released by sending bacteria will be uptaken by the front of receiving bacteria. The complex formed by AHL molecules and CinR proteins will then induce the coloration of the front receiving bacteria.

Let's go further to understand how such evolution of this system is possible with only one strain ?

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