Team:Grenoble/Projet/Intro

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* Diaporama
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<h2>The Project, Mercuro-Coli</h1>
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<h2>Introduction :</h2>
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With industries growth, wastes are accumulating and presence of pollutant and toxic components is becoming an international concern.
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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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<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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<li>Ideally, the device should be a plate carried into a packaging.</li>
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<li>Once unpacked, the sample just has to be added.</li>
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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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<h2>Specifications :</h2>
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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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<li>Operating living cell : E. Coli: BW 25 113</li>
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<li>Only one type of bacteria :</li>
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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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<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" width=600px/></a>
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<li>Comparative measure :</li>
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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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So, an IPTG gradient concentration will be made beforehand in the plate constituting our scale of quantification.
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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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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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</li>
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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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<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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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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<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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<p>So, the location of that boundary represents our point of interest that we will try to visualize.</p>
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<li>A visual result :</li>
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<p>The two different behaviors were chosen in purpose.</p>
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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://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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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.
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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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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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Revision as of 10:50, 25 October 2011

Grenoble 2011, Mercuro-Coli iGEM


The Project, Mercuro-Coli

Introduction :

With industries growth, wastes are accumulating and presence of pollutant and toxic components is becoming an international concern.

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 :

  • Ideally, the device should be a plate carried into a packaging.
  • Once unpacked, the sample just has to be added.
  • 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.

Specifications :

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.

  • Operating living cell : E. Coli: BW 25 113
  • Only one type of bacteria :
  • 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.

  • Comparative measure :

The comparative measure will be made by comparing mercury concentration to IPTG concentration. IPTG concentration will is the reference.

So, an IPTG gradient concentration will be made beforehand in the plate constituting our scale of quantification.

By adding uniformly the polluted sample, two different bacteria behaviors will come up depending on the predominant concentration :

  • On the left side, where the mercury concentration is prevailing, bacteria with a sending 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.
  • On the right, where the IPTG concentration is dominant, bacteria with a receiving behavior will come up. They express CinR protein, a cytoplasmic receptor for Quorum Sensing.

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.

So, the location of that boundary represents our point of interest that we will try to visualize.

  • A visual result :

The two different behaviors were chosen in purpose.

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.

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