Team:Grenoble/Projet/Intro

From 2011.igem.org

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<h2>The Project, Mercuro-Coli</h1>
 
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<h2>Introduction :</h2>
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<h1>The Project, Mercuro-Coli</h1>
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<h2>Introduction</h2>
<p>
<p>
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With industries growth, wastes are accumulating and presence of pollutant and toxic products is becoming an international concern.
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With industrial growth, wastes are accumulating and the presence of pollutant and toxic compounds is becoming an international concern.
</p>
</p>
<p>
<p>
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Our project is oriented in this problematic, Mercuro-Coli is a mercury biosensor which allows to detect and quantify mercury contained into polluted water. Intended to fieldwork studies, the device should be very easy to used :
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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:
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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" class="centerwide"/></a>
 
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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>
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Easy to use in three steps!
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<ul>
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<li>Ideally, the device should be a plate carried into a packaging.</li>
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<li>
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<li>Once unpacked, the sample just has to be added.</li>
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Unpack the plate.
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<li>Finally, the result would be given by a red strip : its presence testifying that the sample contains mercury and its position on the plate acquainting about the quantity.</li>
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</li>
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<li>
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Deposit the polluted water 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>
</ul>
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<h2>Specifications :</h2>
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<h2>Specifications</h2>
<p>
<p>
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At the beginning of the project, specifications were defined about the final device. To get a better understanding on how the device should run, those are going to be taken back step by step.
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At the beginning of the project, we defined a certain number of specifications for the device:
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<ul>
<ul>
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<li>Operating living cell : E. Coli: BW 25 113</li>
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<li>
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<li>Only one type of bacteria :</li>
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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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<p>Once our genetic circuit achieved, it will be introduce to our E. Coli strain. Newly obtained bacteria strain will be then spread on the plate forming a bacteria film on the top of it.</p>
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</li>
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<strong>Form a biofilm on the plate with <i>E. coli</i> bacteria containing the designed genetic circuit.</strong><br/>
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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.  
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<a href="https://static.igem.org/mediawiki/2011/c/c5/1_type_bacteria.png"><img src="https://static.igem.org/mediawiki/2011/c/c5/1_type_bacteria.png" class="centerwide"/></a>
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<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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Plate covered by the bacteria.
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<ul>
<ul>
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<li>Comparative measure :</li>
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<li><strong>Build a comparative measurement system:</strong></li>
</ul>
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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="280px"></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>
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<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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So, an IPTG gradient concentration will be made beforehand constituting our scale of quantification.
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<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:
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<li>
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On the left side, where the mercury concentration is the uppermost, bacteria with a <strong>secretory</strong> behavior will appear. They have the ability to release in the external medium AHL molecules thanks to the expression of 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>
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On the right, where the IPTG concentration is the uppermost, bacteria with a <strong>receiving</strong> behavior will come up. They express internal protein CinR able to receive AHL molecules.
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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>
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</td>
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<td>
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<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="280px"></a>
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<center>
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<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>
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<p>
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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.
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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>
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The key point is therefore the localization of the boundary, which we need to visualize
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<ul>
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<li><strong>A visual result:</strong></li>
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<table class="nobordure">
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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>
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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>
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</center>
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<p>
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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.
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<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>
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<div class="legend">
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Let's go further to understand how such evolution of this system is possible with only one strain ?
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document.getElementById('submenu').innerHTML = '<h3><span class="vert">Le Projet:</span> Bio</h3><ul><li><a href="https://2011.igem.org/Team:Grenoble/Projet/Intro">Introduction</a></li><li><a href="https://2011.igem.org/Team:Grenoble/Projet/Device">The device</a></li><li><a href="https://2011.igem.org/Team:Grenoble/Projet/Design">The genetic circuit:</a></li><ol><li><a href="https://2011.igem.org/Team:Grenoble/Projet/Design/toggle">The toggle switch</a></li><li><a href="https://2011.igem.org/Team:Grenoble/Projet/Design/quorum">The quorum sensing</a></li><li><a href="https://2011.igem.org/Team:Grenoble/Projet/Design/quorum#color">The coloration</a></li><li><a href="https://2011.igem.org/Team:Grenoble/Projet/regulation">Post-transcriptional regulation</a></li></ol><li><a href="https://2011.igem.org/Team:Grenoble/Projet/Modelling">Modelling</a></li><li><a href="https://2011.igem.org/Team:Grenoble/Projet/Results">Results</a></li><li><a href="https://2011.igem.org/Team:Grenoble/Projet/Biobricks">Biobricks</a></li></ul>'
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<h2> Abstract</h2>
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Only ranges of pollutant concentration have been measured in the iGEM competition until now. Our goal is to create a simple and accurate, ready-to-use bioquantifier for heavy metals. Simple enough for an easy visual checking and a routine use in the labs. Our system is inspired from Gardner's work on toggle switch biosystem and from former iGem teams' works on quorum sensing communication.
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Our purpose is to add a pollutant to a device containing a bacterial layer on an engineered medium. This engineered medium is an IPTG gradient.Add of that pollutant to our system induces a geographical division of the plate into two areas : one where the IPTG concentration prevails, and the other where the pollutant concentration prevails. Toggle switch is a biological network which allows to lock our biosystem in a specific state. We use it here in association with complementary quorum sensing genes : A sender and a receiver. Thus, at the interface between both, the reception of quorum sensing molecules by the receptors will induce the coloration of the bacteria. Finally we will obtain a colored line from which we can get the unknown concentration of pollutant.
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Up to now the quantification of heavy metals requires complex physical and chemical protocols. We propose a new way of quantifying heavy metals, much easier than these ones. A first step to a pure water for everyone!
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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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iGEM 2011 Main Page


Contact Us:

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