http://2014.igem.org/wiki/index.php?title=Special:Contributions&feed=atom&limit=20&target=Laureen&year=&month=2014.igem.org - User contributions [en]2024-03-28T18:09:41ZFrom 2014.igem.orgMediaWiki 1.16.5http://2014.igem.org/Team:Toulouse/Result/achievementTeam:Toulouse/Result/achievement2014-10-17T22:46:42Z<p>Laureen: </p>
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<h2>Achievement</h2><br />
<p>Bronze, Silver or Gold Medal? That is the question!</p><br />
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<p style="margin:0 auto; color:#696969; width:960px; padding-top:20px; font-size:16px;"> Results&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Achievement</p> <br />
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<p class="title1" style="text-align:left;">Let's sum up what we did during this summer!</p><br />
<p class="title2">First step...</p><br />
<p class="texte"> <br />
After a summer of hard work, our team is proud of the achieved results. The first step of our project was to have a <b>proof of concept</b> of the three functions requiered to fight against canker. The three modules tested were Chemotaxis, Binding and Fungicides. The verifications of our genetic constructions were successful (PCR, Migration on an agarose gel and Sequencing). We could then proceed to specific test of the modules: capillary test, chitin beads test or fungicide tests.</p><br />
<br />
<p class="title2">Second step...</p><br />
<p class="texte"> <br />
Once these results obtained, we decided to move on to the next step: in order to get closer to our final objective, the three genetic modules have to be assembled and to be tested in <b>model plants</b>. In the short time left that we had, these tests were only performed with the antifungal module, but the results were very encouraging (See <a href="https://2014.igem.org/Team:Toulouse/Result/experimental-results">3. In planta tests with SubtiTree</a> in the Fungicides module). <br />
<br />
<p class="title2">Third step...</p><br />
<p class="texte"> <br />
So encouraging that we moved on to the third step of our project and think about the <b>real injection of SubtiTree in diseased trees</b>. We then considered the <a href="https://2014.igem.org/Team:Toulouse/Project/Spreading"><b>spreading</b></a> problems and proposed different strategies to avoid this major issue. <a href="https://2014.igem.org/Team:Toulouse/Modelling"><b>Modeling</b></a> enables us to predict the growth and the lifespan of SubtiTree in Plane Trees. <br />
<p class="title2">And what comes next?</p><br />
<p class="texte">This project could be carried on as a thesis because the problem of canker killing plane trees is a major concern. Moreover, the support of the Ministry and VNF could help us to continue and achieve our goal: save the plane trees and preserve the beauty of the Canal du Midi.<br><br />
However, SubtiTree dissemination must be controlled, fungicide production regulated, and tests of the whole system performed. We also need to ask that the fungicides chosen could be used according to the legislation. In a nutshell, we still have a lot of things to do even if we did our best during the whole summer!</p><br />
<br />
<p class="title2">iGEM Community</p><br />
<p class="texte"><br />
Well aware that we are part of the <b>iGEM Community</b>, our team also worked for this community by improving the iGEMer's Bible: <a href="http://parts.igem.org/Catalog">the Registry of Standard Biological Parts</a>. <br><br />
We improved the EcAMP BioBrick by adding a promotor, a RBS, but mainly by the addition of a STOP codon (<a href="http://parts.igem.org/Part:BBa_K1364019">BBa_K1364019</a>).<br><br />
As <i>Bacillus subtilis</i> appears to be for us an ideal chassis, we also developed new genetic tools to facilitate its utilization. Thanks to numerous interactions with other iGEM teams, we didn't see the time flying by during this <b>amazing summer</b> !</p><br />
<br />
<center><img style="width:1000px; " src="https://static.igem.org/mediawiki/parts/e/e2/Ach.jpg"></center><br />
<br><br />
<p class="title2">It is better to have "well-made rather than a well-filled head"... (Michel de Montaigne)</p><br />
<p class="texte">We developed a deep <a href="https://2014.igem.org/Team:Toulouse/ethics">ethical questioning</a>, trying to understand if we had the right to act against canker. We strongly believe that neglecting the ethical aspect in a scientific project is a mistake. All the team was involved in this part and the reflexions made were very profitable to us.</p><br />
<br />
<p class="texte">We filled the <a href="https://igem.org/2014_Judging_Form?id=1364">2014 Judging Form</a> thanks to the following accomplishments:</p><br />
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<td style="border-bottom:4px solid #e5e6e6;"><center><br>Silver Medal <img src="https://static.igem.org/mediawiki/2013/d/dd/Medal_silver.png" class="imgcontenttable"/></center><br></td><br />
<td style="border-bottom:4px solid #e5e6e6; border-top-right-radius:9px;"><center><br>Gold Medal<img src="https://static.igem.org/mediawiki/2013/b/bd/Medal_gold_3.png" class="imgcontenttable"/></center><br></td><br />
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<td style="border-right:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Register the team, have a great summer, and plan to have fun at the Jamboree</td><br />
<td style="border-right:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Experimentally validate that at least three new BioBrick Part or Device of your own design and construction work as expected.</td><br />
<td><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Improve the function of an existing BioBrick Part or Device and improve the <i>Bacillus subtilis</i> genetic tools, enter this information in the Registry : <a href="http://parts.igem.org/Part:BBa_K1364019"target="_blank">BBa_K1364019</a>, <a href="http://parts.igem.org/Part:BBa_K1364016"target="_blank">BBa_K1364016</a>, <a href="http://parts.igem.org/Part:BBa_K1364017"target="_blank">BBa_K1364017</a>, <a href="http://parts.igem.org/Part:BBa_K1364021"target="_blank">BBa_K1364021</a>, </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Successfully complete and submit this iGEM Judging form </td><br />
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Document the characterization of these three parts in the 'Main Page' of that Part's/Device's Registry entry<br />
Submit this new part to the iGEM Parts Registry:<br> <a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>, <a href="http://parts.igem.org/Part:BBa_K1364009"target="_blank">BBa_K1364009</a>, <a href="http://parts.igem.org/Part:BBa_K1364013"target="_blank">BBa_K1364013.</a></td><br />
<td style="border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Your project may have implications for the environment, security, safety and ethics and/or ownership and sharing. Describe a novel approach that your team has used to help you and others consider these aspects of the design and outcomes of synthetic biology efforts. → <a href="https://2014.igem.org/Team:Toulouse/ethics" target="_blank">Ethic Part</a> </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Create and share a Description of the team’s project using the iGEM wiki and the team’s parts using the Registry Biological Parts </td><br />
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Submit these new part to the iGEM Parts Registry (submissions must adhere to the iGEM Registry guidelines)</td><br />
<td style="border-top:1px solid #e5e6e6;"> </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Plan to present a Poster and Talk at the iGEM Jamboree </td><br />
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Your project may have implications for the environment, security, safety and ethics and/or ownership and sharing. Describe one or more ways in which these or other broader implications have been taken into consideration in the design and execution of your project. → <a href="https://2014.igem.org/Team:Toulouse/ethics" target="_blank"> Ethical Page</a></td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Document at least one new standard BioBrick Part or Device used in your project/central to your project and submit this part to the iGEM Registry : <a href="http://parts.igem.org/Part:BBa_K1364000" target="_blank">BBa_K1364000</a>, <a href="http://parts.igem.org/Part:BBa_K1364002"target="_blank">BBa_K1364002</a>, <a href="http://parts.igem.org/Part:BBa_K1364003"target="_blank">BBa_K1364003</a>, <a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>. </td><br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/Result/achievementTeam:Toulouse/Result/achievement2014-10-17T22:43:35Z<p>Laureen: </p>
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<h2>Achievement</h2><br />
<p>Bronze, Silver or Gold Medal? That is the question!</p><br />
</div><br />
</div><br />
</div><br />
<br />
<div class="fils-ariane" style="width:100%; height:60px; background:#ededed;"><br />
<p style="margin:0 auto; color:#696969; width:960px; padding-top:20px; font-size:16px;"> Results&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Achievement</p> <br />
</div><br />
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<br />
<div id="innercontenthome"><br />
<div class="centering" style="padding-top: 85px; padding-bottom:40px;"><br />
<br />
<p class="title1" style="text-align:left;">Let's sum up what we did during this summer!</p><br />
<p class="title2">First step...</p><br />
<p class="texte"> <br />
After a summer of hard work, our team is proud of the achieved results. The first step of our project was to have a <b>proof of concept</b> of the three functions requiered to fight against canker. The three modules tested were Chemotaxis, Binding and Fungicides. The verifications of our genetic constructions were successful (PCR, Migration on an agarose gel and Sequencing). We could then proceed to specific test of the modules: capillary test, chitin beads test or fungicide tests.</p><br />
<br />
<p class="title2">Second step...</p><br />
<p class="texte"> <br />
Once these results obtained, we decided to move on to the next step: in order to get closer to our final objective, the three genetic modules have to be assembled and to be tested in <b>model plants</b>. In the short time left that we had, these tests were only performed with the antifungal module, but the results were very encouraging (See <a href="https://2014.igem.org/Team:Toulouse/Result/experimental-results">3. In planta tests with SubtiTree</a> in the Fungicides module). <br />
<br />
<p class="title2">Third step...</p><br />
<p class="texte"> <br />
So encouraging that we moved on to the third step of our project and think about the <b>real injection of SubtiTree in diseased trees</b>. We then considered the <a href="https://2014.igem.org/Team:Toulouse/Project/Spreading"><b>spreading</b></a> problems and proposed different strategies to avoid this major issue. <a href="https://2014.igem.org/Team:Toulouse/Modelling"><b>Modeling</b></a> enables us to predict the growth and the lifespan of SubtiTree in Plane Trees. <br />
<p class="title2">And what comes next?</p><br />
<p class="texte">This project could be carried on as a thesis because the problem of canker killing plane trees is a major concern. Moreover, the support of the Ministry and VNF could help us to continue and achieve our goal: save the plane trees and preserve the beauty of the Canal du Midi.<br><br />
However, SubtiTree dissemination must be controlled, fungicide production regulated, and tests of the whole system performed. We also need to ask that the fungicides chosen could be used according to the legislation. In a nutshell, we still have a lot of things to do even if we did our best during the whole summer!</p><br />
<br />
<p class="title2">iGEM Community</p><br />
<p class="texte"><br />
Well aware that we are part of the <b>iGEM Community</b>, our team also worked for this community by improving the iGEMer's Bible: <a href="http://parts.igem.org/Catalog">the Registry of Standard Biological Parts</a>. <br><br />
We improved the EcAMP BioBrick by adding a promotor, a RBS, but mainly by the addition of a STOP codon (<a href="http://parts.igem.org/Part:BBa_K1364019">BBa_K1364019</a>).<br><br />
As <i>Bacillus subtilis</i> appears to be for us an ideal chassis, we also developed new genetic tools to facilitate its utilization. Thanks to numerous interactions with other iGEM teams, we didn't see the time flying by during this <b>amazing summer</b> !</p><br />
<br />
<center><img style="width:1000px; " src="https://static.igem.org/mediawiki/parts/e/e2/Ach.jpg"></center><br />
<br><br />
<p class="title2">It is better to have "well-made rather than a well-filled head"... (Michel de Montaigne)</p><br />
<p class="texte">We developed a deep <a href="https://2014.igem.org/Team:Toulouse/ethics">ethical questioning</a>, trying to understand if we had the right to act against canker. We strongly believe that neglecting the ethical aspect in a scientific project is a mistake. All the team was involved in this part and the reflexions made were very profitable to us.</p><br />
<br />
<p class="texte">We filled the <a href="https://igem.org/2014_Judging_Form?id=1364">2014 Judging Form</a> with the following achievements:</p><br />
<br />
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<td style="border-bottom:4px solid #e5e6e6; border-top-left-radius:9px;"><center><br>Bronze Medal <img src="https://static.igem.org/mediawiki/2013/8/86/Medal_bronze.png" class="imgcontenttable"/></center><br></td><br />
<td style="border-bottom:4px solid #e5e6e6;"><center><br>Silver Medal <img src="https://static.igem.org/mediawiki/2013/d/dd/Medal_silver.png" class="imgcontenttable"/></center><br></td><br />
<td style="border-bottom:4px solid #e5e6e6; border-top-right-radius:9px;"><center><br>Gold Medal<img src="https://static.igem.org/mediawiki/2013/b/bd/Medal_gold_3.png" class="imgcontenttable"/></center><br></td><br />
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<td style="border-right:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Register the team, have a great summer, and plan to have fun at the Jamboree</td><br />
<td style="border-right:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Experimentally validate that at least three new BioBrick Part or Device of your own design and construction work as expected.</td><br />
<td><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Improve the function of an existing BioBrick Part or Device and improve the <i>Bacillus subtilis</i> genetic tools, enter this information in the Registry : <a href="http://parts.igem.org/Part:BBa_K1364019"target="_blank">BBa_K1364019</a>, <a href="http://parts.igem.org/Part:BBa_K1364016"target="_blank">BBa_K1364016</a>, <a href="http://parts.igem.org/Part:BBa_K1364017"target="_blank">BBa_K1364017</a>, <a href="http://parts.igem.org/Part:BBa_K1364021"target="_blank">BBa_K1364021</a>, </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Document the characterization of these three parts in the 'Main Page' of that Part's/Device's Registry entry<br />
Submit this new part to the iGEM Parts Registry:<br> <a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>, <a href="http://parts.igem.org/Part:BBa_K1364009"target="_blank">BBa_K1364009</a>, <a href="http://parts.igem.org/Part:BBa_K1364013"target="_blank">BBa_K1364013.</a></td><br />
<td style="border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Your project may have implications for the environment, security, safety and ethics and/or ownership and sharing. Describe a novel approach that your team has used to help you and others consider these aspects of the design and outcomes of synthetic biology efforts. → <a href="https://2014.igem.org/Team:Toulouse/ethics" target="_blank">Ethic Part</a> </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Create and share a Description of the team’s project using the iGEM wiki and the team’s parts using the Registry Biological Parts </td><br />
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Submit these new part to the iGEM Parts Registry (submissions must adhere to the iGEM Registry guidelines)</td><br />
<td style="border-top:1px solid #e5e6e6;"> </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Plan to present a Poster and Talk at the iGEM Jamboree </td><br />
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Your project may have implications for the environment, security, safety and ethics and/or ownership and sharing. Describe one or more ways in which these or other broader implications have been taken into consideration in the design and execution of your project. → <a href="https://2014.igem.org/Team:Toulouse/ethics" target="_blank"> Ethical Page</a></td><br />
<td style="border-top:1px solid #e5e6e6;"></td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Document at least one new standard BioBrick Part or Device used in your project/central to your project and submit this part to the iGEM Registry : <a href="http://parts.igem.org/Part:BBa_K1364000" target="_blank">BBa_K1364000</a>, <a href="http://parts.igem.org/Part:BBa_K1364002"target="_blank">BBa_K1364002</a>, <a href="http://parts.igem.org/Part:BBa_K1364003"target="_blank">BBa_K1364003</a>, <a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>. </td><br />
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<!-------------------------------- FOOTER ---------------------------------><br />
{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/Result/achievementTeam:Toulouse/Result/achievement2014-10-17T22:38:20Z<p>Laureen: </p>
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<h2>Achievement</h2><br />
<p>Bronze, Silver or Gold Medal? That is the question!</p><br />
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<p style="margin:0 auto; color:#696969; width:960px; padding-top:20px; font-size:16px;"> Results&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Achievement</p> <br />
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<p class="title1" style="text-align:left;">Let's sum up what we did during this summer!</p><br />
<p class="title2">First step...</p><br />
<p class="texte"> <br />
After a summer of hard work, our team is proud of the achieved results. The first step of our project was to have a <b>proof of concept</b> of the three functions requiered to fight against canker. The three modules tested were Chemotaxis, Binding and Fungicides. The verifications of our genetic constructions were successful (PCR, Migration on an agarose gel and Sequencing). We could then proceed to specific test of the modules: capillary test, chitin beads test or fungicide tests.</p><br />
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<p class="title2">Second step...</p><br />
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Once these results obtained, we decided to move on to the next step: in order to get closer to our final objective, the three genetic modules have to be assembled and to be tested in <b>model plants</b>. In the short time left that we had, these tests were only performed with the antifungal module, but the results were very encouraging (See <a href="https://2014.igem.org/Team:Toulouse/Result/experimental-results">3. In planta tests with SubtiTree</a> in the Fungicides module). <br />
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<p class="title2">Third step...</p><br />
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So encouraging that we moved on to the third step of our project and think about the <b>real injection of SubtiTree in diseased trees</b>. We then considered the <a href="https://2014.igem.org/Team:Toulouse/Project/Spreading"><b>spreading</b></a> problems and proposed different strategies to avoid this major issue. <a href="https://2014.igem.org/Team:Toulouse/Modelling"><b>Modeling</b></a> enables us to predict the growth and the lifespan of SubtiTree in Plane Trees. <br />
<p class="title2">And what comes next?</p><br />
<p class="texte">This project could be carried on as a thesis because the problem of canker killing plane trees is a major concern. Moreover, the support of the Ministry and VNF could help us to continue and achieve our goal: save the plane trees and preserve the beauty of the Canal du Midi.<br><br />
However, SubtiTree dissemination must be controlled, fungicide production regulated, and tests of the whole system performed. We also need to ask that the fungicides chosen could be used according to the legislation. In a nutshell, we still have a lot of things to do even if we did our best during the whole summer!</p><br />
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<p class="title2">iGEM Community</p><br />
<p class="texte"><br />
Well aware that we are part of the <b>iGEM Community</b>, our team also worked for this community by improving the iGEMer's Bible: <a href="http://parts.igem.org/Catalog">the Registry of Standard Biological Parts</a>. <br><br />
We improved the EcAMP BioBrick by adding a promotor, a RBS, but mainly by the addition of a STOP codon (<a href="http://parts.igem.org/Part:BBa_K1364019">BBa_K1364019</a>).<br><br />
As <i>Bacillus subtilis</i> appears to be for us an ideal chassis, we also developed new genetic tools to facilitate its utilization. Thanks to numerous interactions with other iGEM teams, we didn't see the time flying by during this <b>amazing summer</b> !</p><br />
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<center><img style="width:1000px; " src="https://static.igem.org/mediawiki/parts/e/e2/Ach.jpg"></center><br />
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<p class="title2">It is better to have "well-made rather than a well-filled head"... (Michel de Montaigne)</p><br />
<p class="texte">We developed a deep <a href="https://2014.igem.org/Team:Toulouse/ethics">ethical questioning</a>, trying to understand if we had the right to act against canker. We strongly believe that neglecting the ethical aspect in a scientific project is a mistake. All the team was involved in this part and the reflexions made were very profitable to us.</p><br />
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<p class="title1" style="text-align:left;">Medals Fulfillment</p> <br />
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<td style="border-bottom:4px solid #e5e6e6; border-top-left-radius:9px;"><center><br>Bronze Medal <img src="https://static.igem.org/mediawiki/2013/8/86/Medal_bronze.png" class="imgcontenttable"/></center><br></td><br />
<td style="border-bottom:4px solid #e5e6e6;"><center><br>Silver Medal <img src="https://static.igem.org/mediawiki/2013/d/dd/Medal_silver.png" class="imgcontenttable"/></center><br></td><br />
<td style="border-bottom:4px solid #e5e6e6; border-top-right-radius:9px;"><center><br>Gold Medal<img src="https://static.igem.org/mediawiki/2013/b/bd/Medal_gold_3.png" class="imgcontenttable"/></center><br></td><br />
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<td style="border-right:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Register the team, have a great summer, and plan to have fun at the Jamboree</td><br />
<td style="border-right:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Experimentally validate that at least three new BioBrick Part or Device of your own design and construction work as expected.</td><br />
<td><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Improve the function of an existing BioBrick Part or Device and improve the <i>Bacillus subtilis</i> genetic tools, enter this information in the Registry : <a href="http://parts.igem.org/Part:BBa_K1364019"target="_blank">BBa_K1364019</a>, <a href="http://parts.igem.org/Part:BBa_K1364016"target="_blank">BBa_K1364016</a>, <a href="http://parts.igem.org/Part:BBa_K1364017"target="_blank">BBa_K1364017</a>, <a href="http://parts.igem.org/Part:BBa_K1364021"target="_blank">BBa_K1364021</a>, </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Successfully complete and submit this iGEM Judging form </td><br />
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Document the characterization of these three parts in the 'Main Page' of that Part's/Device's Registry entry<br />
Submit this new part to the iGEM Parts Registry:<br> <a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>, <a href="http://parts.igem.org/Part:BBa_K1364009"target="_blank">BBa_K1364009</a>, <a href="http://parts.igem.org/Part:BBa_K1364013"target="_blank">BBa_K1364013.</a></td><br />
<td style="border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Your project may have implications for the environment, security, safety and ethics and/or ownership and sharing. Describe a novel approach that your team has used to help you and others consider these aspects of the design and outcomes of synthetic biology efforts. → <a href="https://2014.igem.org/Team:Toulouse/ethics" target="_blank">Ethic Part</a> </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Create and share a Description of the team’s project using the iGEM wiki and the team’s parts using the Registry Biological Parts </td><br />
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Submit these new part to the iGEM Parts Registry (submissions must adhere to the iGEM Registry guidelines)</td><br />
<td style="border-top:1px solid #e5e6e6;"> </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Plan to present a Poster and Talk at the iGEM Jamboree </td><br />
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Your project may have implications for the environment, security, safety and ethics and/or ownership and sharing. Describe one or more ways in which these or other broader implications have been taken into consideration in the design and execution of your project. → <a href="https://2014.igem.org/Team:Toulouse/ethics" target="_blank"> Ethical Page</a></td><br />
<td style="border-top:1px solid #e5e6e6;"></td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Document at least one new standard BioBrick Part or Device used in your project/central to your project and submit this part to the iGEM Registry : <a href="http://parts.igem.org/Part:BBa_K1364000" target="_blank">BBa_K1364000</a>, <a href="http://parts.igem.org/Part:BBa_K1364002"target="_blank">BBa_K1364002</a>, <a href="http://parts.igem.org/Part:BBa_K1364003"target="_blank">BBa_K1364003</a>, <a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>. </td><br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/Result/achievementTeam:Toulouse/Result/achievement2014-10-17T22:34:20Z<p>Laureen: </p>
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<h2>Achievement</h2><br />
<p>Bronze, Silver or Gold Medal? That is the question!</p><br />
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<p style="margin:0 auto; color:#696969; width:960px; padding-top:20px; font-size:16px;"> Results&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Achievement</p> <br />
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<p class="title1" style="text-align:left;">Let's sum up what we did during this summer!</p><br />
<p class="title2">First step...</p><br />
<p class="texte"> <br />
After a summer of hard work, our team is proud of the achieved results. The first step of our project was to have a <b>proof of concept</b> of the three functions requiered to fight against canker. The three modules tested were Chemotaxis, Binding and Fungicides. The verifications of our genetic constructions were successful (PCR, Migration on an agarose gel and Sequencing). We could then proceed to specific test of the modules: capillary test, chitin beads test or fungicide tests.</p><br />
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<p class="title2">Second step...</p><br />
<p class="texte"> <br />
Once these results obtained, we decided to move on to the next step: in order to get closer to our final objective, the three genetic modules have to be assembled and to be tested in <b>model plants</b>. In the short time left that we had, these tests were only performed with the antifungal module, but the results were very encouraging (See <a href="https://2014.igem.org/Team:Toulouse/Result/experimental-results">3. In planta tests with SubtiTree</a> in the Fungicides module). <br />
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<p class="title2">Third step...</p><br />
<p class="texte"> <br />
So encouraging that we moved on to the third step of our project and think about the <b>real injection of SubtiTree in diseased trees</b>. We then considered the <a href="https://2014.igem.org/Team:Toulouse/Project/Spreading"><b>spreading</b></a> problems and proposed different strategies to avoid this major issue. <a href="https://2014.igem.org/Team:Toulouse/Modelling"><b>Modeling</b></a> enables us to predict the growth and the lifespan of SubtiTree in Plane Trees. <br />
<p class="title2">And what comes next?</p><br />
<p class="texte">This project could be continued thanks to a thesis as the problem of canker killing plane trees is a major concern. Moreover, the support of the Ministry and VNF could help us to continue and achieve our goal: save the plane trees and preserve the beauty of the Canal du Midi.<br><br />
However, SubtiTree dissemination must be controlled, fungicide production regulated, and tests of the whole system performed. We also need to ask that the fungicides chosen could be used according to the legislation. In a nutshell, we still have a lot of things to do even if we did our best during all the summer!</p><br />
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<p class="title2">iGEM Community</p><br />
<p class="texte"><br />
Well aware that we are part of the <b>iGEM Community</b>, our team also worked for this community by improving the iGEMer's Bible: <a href="http://parts.igem.org/Catalog">the Registry of Standard Biological Parts</a>. <br><br />
We improved the EcAMP BioBrick by adding a promotor, a RBS, but mainly by the addition of a STOP codon (<a href="http://parts.igem.org/Part:BBa_K1364019">BBa_K1364019</a>).<br><br />
As <i>Bacillus subtilis</i> appears to be for us an ideal chassis, we also developed new genetic tools to facilitate its utilization. Thanks to numerous interactions with other iGEM teams, we didn't see the time flying by during this <b>amazing summer</b> !</p><br />
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<p class="title2">It is better to have "well-made rather than a well-filled head"... (Michel de Montaigne)</p><br />
<p class="texte">We developed a deep <a href="https://2014.igem.org/Team:Toulouse/ethics">ethical questioning</a>, trying to understand if we had the right to act against canker. We strongly believe that neglecting the ethical aspect in a scientific project is a mistake. All the team was involved in this part and the reflexions made were very profitable to us.</p><br />
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<td style="border-bottom:4px solid #e5e6e6;"><center><br>Silver Medal <img src="https://static.igem.org/mediawiki/2013/d/dd/Medal_silver.png" class="imgcontenttable"/></center><br></td><br />
<td style="border-bottom:4px solid #e5e6e6; border-top-right-radius:9px;"><center><br>Gold Medal<img src="https://static.igem.org/mediawiki/2013/b/bd/Medal_gold_3.png" class="imgcontenttable"/></center><br></td><br />
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<td style="border-right:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Register the team, have a great summer, and plan to have fun at the Jamboree</td><br />
<td style="border-right:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Experimentally validate that at least three new BioBrick Part or Device of your own design and construction work as expected.</td><br />
<td><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Improve the function of an existing BioBrick Part or Device and improve the <i>Bacillus subtilis</i> genetic tools, enter this information in the Registry : <a href="http://parts.igem.org/Part:BBa_K1364019"target="_blank">BBa_K1364019</a>, <a href="http://parts.igem.org/Part:BBa_K1364016"target="_blank">BBa_K1364016</a>, <a href="http://parts.igem.org/Part:BBa_K1364017"target="_blank">BBa_K1364017</a>, <a href="http://parts.igem.org/Part:BBa_K1364021"target="_blank">BBa_K1364021</a>, </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/>Document the characterization of these three parts in the 'Main Page' of that Part's/Device's Registry entry<br />
Submit this new part to the iGEM Parts Registry:<br> <a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>, <a href="http://parts.igem.org/Part:BBa_K1364009"target="_blank">BBa_K1364009</a>, <a href="http://parts.igem.org/Part:BBa_K1364013"target="_blank">BBa_K1364013.</a></td><br />
<td style="border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Your project may have implications for the environment, security, safety and ethics and/or ownership and sharing. Describe a novel approach that your team has used to help you and others consider these aspects of the design and outcomes of synthetic biology efforts. → <a href="https://2014.igem.org/Team:Toulouse/ethics" target="_blank">Ethic Part</a> </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Create and share a Description of the team’s project using the iGEM wiki and the team’s parts using the Registry Biological Parts </td><br />
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Submit these new part to the iGEM Parts Registry (submissions must adhere to the iGEM Registry guidelines)</td><br />
<td style="border-top:1px solid #e5e6e6;"> </td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Plan to present a Poster and Talk at the iGEM Jamboree </td><br />
<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Your project may have implications for the environment, security, safety and ethics and/or ownership and sharing. Describe one or more ways in which these or other broader implications have been taken into consideration in the design and execution of your project. → <a href="https://2014.igem.org/Team:Toulouse/ethics" target="_blank"> Ethical Page</a></td><br />
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<td style="border-right:1px solid #e5e6e6; border-top:1px solid #e5e6e6;"><img style="width:20px;" src="https://static.igem.org/mediawiki/2014/3/34/Career-check.jpg" class="imgcontenttableleft"/> Document at least one new standard BioBrick Part or Device used in your project/central to your project and submit this part to the iGEM Registry : <a href="http://parts.igem.org/Part:BBa_K1364000" target="_blank">BBa_K1364000</a>, <a href="http://parts.igem.org/Part:BBa_K1364002"target="_blank">BBa_K1364002</a>, <a href="http://parts.igem.org/Part:BBa_K1364003"target="_blank">BBa_K1364003</a>, <a href="http://parts.igem.org/Part:BBa_K1364005"target="_blank">BBa_K1364005</a>. </td><br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/ethicsTeam:Toulouse/ethics2014-10-17T22:06:59Z<p>Laureen: </p>
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<p style="color:#696969; padding-top:20px; font-size:16px; float:left;"> Human practice&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Ethics</p> <br />
</div> <br />
</div><br />
<br />
<div id="innercontenthome"><br />
<div class="centering" style="padding-top: 85px; padding-bottom:40px;"><br />
<br />
<div id="column-left"><br />
<h3 class="title2" style="margin-top:10px; color:#333;">Summary :</h3><br />
<ul class="menuleft"><br />
<li style="margin-top:25px;"><a href="#select1">Protection of the beauty</a></li><br />
<li><a href="#select2">Human intervention in the nature</a></li><br />
<li><a href="#select3">SubtiTree</a></li><br />
</ul><br />
</div><br />
<br />
<div class="column-right" style="width:75%; float:right;"><br />
<br />
<!--CITATION--><br />
<p class="citation"><br />
"Ethics is as important as laws."<br />
</p><br />
<br />
<p class="texte">The ethical<br />
questioning turned out to be one of the major starting points of our project.<br />
Acting on an established environment and modifying it is by no mean trivial and<br />
our combined technical and philosophical points of view. The actual purpose of our project also leads us to undertake an<br />
ethical questioning about the role of the scientist regarding “useless” things<br />
such as the trees lining along the Canal du Midi. </p><br />
<br />
<p class="title1" id="select1">Protection of the beauty</p><br />
<p class="title2">Is it the scientists’ role to protect beauty?<br />
</p><br />
<br />
<p class="texte"> Beauty is a<br />
feeling of satisfaction and is selfless. It is more a feeling than the property<br />
of a thing, this is not a notion we can clearly understand. Indeed, we can find<br />
something beautiful even when we don’t know the purpose of the object...</p><br />
<center><br />
<img src="https://static.igem.org/mediawiki/2014/f/fa/Fontaine_Duchamp.jpg" width="400px"><br />
<p class="legend">Figure 1: Fontaine (Marcel Duchamp). Yes this is also art...</p></center><br />
<br />
<p class="texte">There is always<br />
a distinction between natural beauty and artistic beauty according to Hegel,<br />
the famous philosopher. The artistic beauty is born from our mind and our<br />
spirit: it is an element of signification of the work of art whereas the<br />
natural beauty of the object is external. In a way, the Canal du Midi combines<br />
both types of beauty: a natural one regarding the Nature, the centenary plane<br />
trees but also an artistic one since the Canal was built by the human hands.<br />
Usually, science judges beauty as a superficial feature not deserving to<br />
undertake any kind of scientific efforts to maintain it. The traditional role<br />
of science is to solve global issues and to elaborate complex strategies in<br />
order to find useful solutions for everyone’s life. Once made this observation,<br />
one may wonder why synthetic biology would be used only to protect the useless<br />
beauty of a local heritage such as the trees lining the Canal du Midi. <br />
</p><br />
<br />
<p class="texte"><br />
This crucial<br />
interrogation leads us to consider science and synthetic biology from a<br />
different point of view. <b>What if the role of scientists was also to make<br />
people rediscovering the beauty of Nature? What if the bases of new scientific<br />
challenges resulted from a more local scale? </b>Science does not have to be it has so much to gain opening itself to these<br />
challenges. First scientifically, as research is never<br />
useless and as we never know the impact and the scope of our results.<br />
Then socially, as we could measure the deep interest raised by our<br />
project within the population and the media. Adopting a new vision of synthetic<br />
biology, we will probably make people change their mind about this innovative<br />
discipline. <br><br />
The traditional cold objectivity of science distances itself from the society.<br />
However, scientists are also being capable of feeling the beauty, sensitive to<br />
the charm of landscapes and <b>able to understand the usefulness of<br />
"useless" trees</b>…<br><br />
The design of a strategy to protect useless beauty may seem senseless but we<br />
believe that it is also the scientist’s duty. We have to remember that thinking<br />
is what distinguish <i>Homo sapiens</i> from other species on earth and this<br />
"thinking" feature allows us to understand the world and be conscious<br />
of our human Nature (Descartes: <i>Cogito ergo sum</i>). The art is an object<br />
of philosophical thought. Consciousness raises humans above all others living<br />
creatures. Thus, it is necessary to respect and protect art. And thus, it<br />
becomes essential to preserve the beauty of the Canal du Midi. <br />
</p><br />
<br />
<br />
<p class="title1" id="select2">Human<br />
intervention on Nature</p><br />
<p class="texte">Our main<br />
question is to understand the complicated relationship between man and Nature.<br />
Does mankind have the proper right to change the Nature? Is modified Nature<br />
considered as artificial?</p><br />
<p class="title2"> Mankind & Nature</p><br />
<br />
<p class="texte"> Nature deserves<br />
to be respected and loved. Mankind has always been linked to Nature as its<br />
survival depends on what comes out of the ground, the trees, the oceans… The<br />
Nature is a source of wealth for mankind. It ensures survival and development<br />
by giving men the wood, the rocks, the soil to build shelters. Being in contact<br />
with Nature can allow men to feel strong emotion, as describe by poets like<br />
Hugo and Lamartine.</p><br />
<br />
<br />
<p class="texte">Since the birth<br />
of humanity, man himself understood the importance of studying and mastering<br />
Nature to develop the civilization. Still today the most advanced technologies<br />
often try to mimic natural phenomena. With the development of civilization, men<br />
modified their environment, changing it for their own comfort depending on<br />
their own desire. With the increase of human activity, the natural environment<br />
is modified profundly. With industrialization, the<br />
natural environment suffered from waste discharges, oil slicks, intensive<br />
fishing (and many others...) but also the introduction of devastating species<br />
such as the pathogen,<i> Ceratocystis<br />
platani</i>. However, despite these negative aspects, men<br />
are capable of favorable actions to help the environment and fix their<br />
mistakes. The current trend is to limit the impact of human interventions on<br />
Nature, and hopefully this trend is not transient and will not vanish. A new<br />
desire is born, a wish to protect Nature and wilderness. Humanity can adhere to<br />
this position: human take advantage of the<br />
environment and the environment takes advantage of the reasoned human<br />
interventions. There is an adaptation of mankind to Nature. Moreover, humans<br />
can have empathy: people are capable of understanding emotions and cognitive<br />
states of other organisms. To respond to these feelings, humans have<br />
technological tools allowing them to fight against enemies. This is the case<br />
with our project: fighting <i>Ceratocystis<br />
platani</i>. <br />
</p><br />
<br />
<p class="texte">In conclusion,<br />
by destroying and hammering the Nature, we jeopardize our lives. We need<br />
Nature, we come from Nature and we depend on Nature for survival, food,<br />
discoveries and civilisation. Respecting, loving and<br />
preserving the beauty of it is also a question of<br />
survival.</p><br />
<br />
<p class="title2">Nature and artifice<br />
</p><br />
<br />
<p class="texte">Talking about<br />
the Nature refers to the whole world with an exception: all the transformations<br />
made by mankind. Nature exists regardless of men and his interventions whereas<br />
artificial is everything that exists because to humans.<br />
</p><br />
<br />
<p class="texte">However,<br />
pretending that natural and artificial are opposite does not seem to be true.<br />
Man cannot create without the various elements provided by Nature, he then justs transform Nature. Thus we may wonder if there is a<br />
true difference between natural and artificial. The border between these two<br />
notions is not as obvious as it seems. The landscapes are shaped by the hand of<br />
man, animals are domesticated, and now bacteria are considered as cell<br />
factories. A natural reserve is artificially preserved as a result of human<br />
actions. Is there still something natural since the birth of mankind? Actually,<br />
the artifice is a slight modification of Nature and couldn’t exist by itself.<br />
The distinction between natural and artificial seems sterile and we clearly<br />
understand that these notions are inextricably linked and need each other to<br />
exist. </p><br />
<br />
<p class="texte">In conclusion,<br />
isn't it our duty to use our unique position in the history of life and our human<br />
approach to try to replace the evolutive processes?</p><br />
<br />
<br />
<p class="title2">Back to our project</p><br />
<br />
<br />
<p class="texte">These<br />
inextricable links are obviously the basis of our project. We aim to<br />
artificially preserve a natural heritage shaped by Pierre Paul Riquet hundreds years ago.Fighting a<br />
naturally occurring form of life that threatens it maybe just an imitation of<br />
the natural evolution process. What is considered today as ‘non-natural’<br />
may be one day regarded differently. To the extent that everything is done not<br />
to unbalance the ecosystem, our intervention can be judged rightful, even more<br />
than the use of chemicals.</p><br />
<br />
<br />
<p class="title1" id="select3">SubtiTree</p><br />
<br />
<p class="title2"> Potential strategies discussed<br />
<br> (See more details in the <a href="https://2014.igem.org/Team:Toulouse/Project/Spreading">Spreading</a> dedicated page)<br />
</p><br />
<br />
<p class="texte">To be sure that<br />
SubtiTree will not survive and spread in the<br />
environment, many strategies were discussed to improve our bacterium: <br />
<br />
<br>- Avoid the survival in the natural environment (outside the tree) thanks to a proline auxotrophy system <br />
<br>- Prevent the sporulation of <i>B. subtilis</i> to make it annual <br />
<br>- Avoid gene transfers between SubtiTree and a wild<br />
type bacterium thanks to a toxin-antitoxin system <br />
<br>- Use an integrative plasmid to improve the genetic stability<br />
</p><br />
<br />
<br />
<br />
<p class="title2">Public perception<br />
</p><br />
<br />
<p class="texte"><I><CENTER>Political and public adhesion</I></CENTER></p><br />
<br />
<p class="texte">Due to our<br />
strong implication in preserving this magnificent work of art, our project<br />
interested several governmental services. Indeed some municipalities and<br />
regional councils supported our local engagement. Beyond that, our project<br />
interests the highest level of the “Canal du Midi” administration: the national<br />
navigation authority (VNF) and the Ministry of agriculture. Both of them funded<br />
this project. They are now looking for the continuation of the project after<br />
the iGEM competition. This is clearly a sign that we targeted the right<br />
question. </p><br />
<br />
<p class="texte">This project<br />
also received the attention of the public through several articles in<br />
newspapers, television, radio and internet. First we had just a local coverage,<br />
but days after days there were more and more media interested in SubtiTree. This mediatic coverage<br />
allowed us to contact concerned citizens who participated to the development of<br />
this project. This interaction with the public allowed us to explain and<br />
promote public knowledge of synthetic biology. </p><br />
<br />
<br />
<p class="texte"><I><CENTER>Safety principle</I></CENTER></p><br />
<br />
<p class="texte">One single tree<br />
infected by Canker, and all the trees located in an area of a couple of hundred<br />
meters around are included in the prophylactic cut. We acted to preserve the<br />
surrounding trees. The modification of the endophytic<br />
microbial fauna generated by the introduction of the engineered bacterium has<br />
to be compared to the introduction of chemicals. They contain chlorine atom and<br />
aromatic hydrocarbon, so their remediation is complicated and they represent a<br />
source of pollution. By shortening the lifespan to one season and minimizing<br />
the risks of spreading, we plan a safe and environmental-friendly way to fight<br />
Canker. <br />
<br />
<br />
<p class="title2">Feasability<br />
</p><br />
<br />
<p class="texte">We wonder about<br />
the feasibility of tree’s treatment. As we used endophytic<br />
bacteria, we can count on the natural growth of SubtiTree<br />
inside the sap. So we can inject few bacteria to be sure to have enough<br />
bacteria to protect the tree. Some researchers (Xianling<br />
Ji<sup>1</sup> et al) already injected <i>Bacillus subtilis</i> in plants and<br />
observe an increase of bacteria concentration to a maximum of 10<sup>5</sup><br />
bacteria/mL</p><br />
<br />
<p class="texte">As we aim to<br />
inject a small quantity of bacteria, this treatment remains cheaper than the<br />
injection of several liters of chemical fungicides. In addition, this injection<br />
prevents the preventive tree cutting, which is very expensive. Cutting one tree<br />
cost around € 3000. The administration in charge of the protection of the<br />
“Canal du Midi” already plans to spend 220 million euros to cut and replant all<br />
trees along the Canal. Besides the important cost of cutting trees, it will<br />
destroy one of the symbols of south-western France. </p><br />
<br />
<p class="texte">We know that SubtiTree could be improved in many ways, but in the <br />
iGEM’s circumstances we could not have the time to go<br />
deeper. First, we can improve the fixation module. Using chitin as fixation<br />
anchor is simple but not enough specific to fix just one fungus type. That’s<br />
why we first think to fix SubtiTree to one protein<br />
included in the <i>Ceratocystis<br />
platani</i>’s<br />
membrane: CP. The bacterial prototype designed this summer can be optimized to<br />
trigger the fungicides production when the binding is completed, and to be more<br />
specific changing the peptides produced.</p><br />
<br />
<p class="title1">References</p><br />
<br />
<li class="tree"><p class="texte"> Xianling Ji, Guobing Lu, Yingping Gai, Chengchao Zheng & Zhimei Mu.<b> Biological control against bacterialwilt and colonization of<br />
mulberry byan endophyticBacillus subtilis strain </b>. FEMS Microbiol Ecol. 65 (2008) 565–573. </p></li><br />
</div><br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/ModellingTeam:Toulouse/Modelling2014-10-17T21:53:30Z<p>Laureen: </p>
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<h2>Modeling</h2><br />
<p>To develop a predictive model</p><br />
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<div class="fils-ariane" style="width:100%; height:60px; background:#ededed;"><br />
<p style="margin:0 auto; color:#696969; width:960px; padding-top:20px; font-size:16px;"> Project&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Modeling</p> <br />
</div><br />
<br />
<br />
<div id="innercontenthome"><br />
<div class="centering" style="padding-top: 85px; padding-bottom:40px;"><br />
<br />
<p class="texte"><br />
<br />
Modeling is a tool used to simplify and study systems. It helps us to predict behavior thanks to bibliographic or experimental informations.</br><br />
<br />
The following modelisation focuses on the development of our engineered bacterium (called SubtiTree) in trees. The bacterial growth in trees seems to be unknown, thus we must infer <i>Bacillus subtilis'</i> behavior.</p><br />
<br />
<p class="title1"><br />
Bacterial Growth<br />
</p><br />
<br />
<p class="title2"><br />
Aim<br />
</p><br />
<br />
<p class="texte"><br />
<i>Bacillus subtilis</i> is a tree endophyte strain. A study showed that <i>Bacillus subtilis</i> could develop and fully colonize a tree, reaching a concentration of 10⁵ cells per gram of fresh plant. We need to know in which conditions the growth of <i>B. subtilis</i> is optimum in a tree and if the weather can stop its development during winter. Therefore we decided to work on the <i>Bacillus subtilis'</i> growth in function of the temperature during the year. <br />
<br>Modeling bacterial growth in a tree section generates some difficulties. We need to know the distance between two tree extremities (treetops and root) or the speed sap flow. However the speed sap flow can vary with temperature during the day and seasons cause of the type of sap (phloem, xylem). Furthermore a tree is not an homogeneous system: its roots, trunk and branches do not contain the same amount of sap and wood. <br>The average speed of the plane tree sap is 2.4m/h, which means that in a day the sap of a 30m tree will flow from one extremity to the other. Tree is reduced to a bioreactor.<br />
</p><br />
<p class="texte"><br />
We make the following hypothesis:</p><br />
<ol class="list1"><br />
<li><br />
According to the publication of <b>Xianling Ji</b> (See References), after six months of <i>Bacillus subtilis</i> growth in a tree, bacteria cells reach a concentration of 10⁵ cells per gram of fresh plant. We assume that 10⁵ cells/g is the maximum concentration.<br />
</li><br />
<li><br />
The composition of the phloem is stable. There is no effect of depletion of the medium.<br />
</li><br />
<li><br />
Only temperature impacts on bacterial growth.<br />
</li><br />
<li><br />
It is assumed that there is no leakage of cells.<br />
</li><br />
</ol><br />
<br />
<br />
<p class="title2"><br />
Method<br />
</p><br />
<br />
<p class="texte"><br />
An assessment of the <i>Bacillus subtilis</i> growth in a similar sap was performed in laboratory conditions with optimum growth medium for <i>Bacillus subtilis</i>. The composition sap used was the one from birch sap.<br><br />
In these conditions, the growth rate μ is optimal. From this value we can extrapolate a growth curve as a function of temperature. We used the <b>cardinal temperature model</b>: </p><br />
<br />
<center style="margin-bottom:50px;"><img style="" src="https://static.igem.org/mediawiki/2014/8/85/Formules_Rosso.png" alt="cardinal temperature model"></center><br />
<br />
<br />
<p class="texte"><br />
T: Temperature</br><br />
µ<sub>opt</sub>: Optimal growth rate</br><br />
µ: growth rate at temperature T</br><br />
T<sub>max</sub>: Maximum temperature supported by bacteria</br><br />
T<sub>min</sub>: Minimum temperature supported by bacteria</br><br />
T<sub>opt</sub>: Optimum temperature for the growth</br></br><br />
<br />
Necessary parameters for this function are minimun temperature T<sub>min</sub> and maximum temperature T<sub>max</sub>, optimal temperature for the growth T<sub>opt</sub> and optimal growth rate µ<sub>opt</sub>.</br><br />
</br><br />
T<sub>min</sub>: 10°C</br><br />
T<sub>max</sub>: 52°C</br><br />
T<sub>opt</sub>: 37°C</br><br />
µ<sub>opt</sub>: 8.5968 cfu/d</br></br><br />
<br />
The optimal growth rate (µ<sub>opt</sub>) is obtained experimentally with a similar birch sap environment.</br><br />
The growth rate is negative below 10°C (according to growth tests performed at 10°C and 4°C under similar conditions for the measurement of μ<sub>opt</sub>), survival rate after 24h was 0.3 % at 10°C and null at 4°C.<br><br />
Conditions apply:</p><br />
<br />
<p class="texte"> <br />
If<span style="color:#FFFFFF; font-family:'Open Sans'; font-size:14px;">__</span>| T<= 4°C -> µ = -1</br><br />
<span style="color:#FFFFFF; font-family:'Open Sans'; font-size:14px;">____</span>| 4°C<T<= 10°C -> µ = -0.97</br><br />
<span style="color:#FFFFFF; font-family:'Open Sans'; font-size:14px;">____</span>| T > 10°C -> µ = f(T) with f(T) egal to cardinal temperature model.</p><br />
<br />
<center style="margin-top: -52px;"><img style="" src="https://static.igem.org/mediawiki/2014/b/b1/Plot_growth_rate.png" alt="Figure1"></center><br />
<p class="legend">Figure 1: bacterial growth (µ) as a function of temperature</p><br />
<br />
<p class="texte"> A logistic model developed by <b>Hiroshi Fujikawa</b> (See References) is used to study bacterial growth.</p><br />
<br />
<p class="legend">General logistics formulas:</p><br />
<center style="margin:-44px 0 65px;"><img style="" src="https://static.igem.org/mediawiki/2014/c/c3/Form_general_fonction.png" alt="General logistics formulas"></center><br />
<br />
<br />
<p class="texte"><br />
In our case, the growth rate µ depends on the temperature. <br />
<br>N corresponds to the bacterial population, N<sub>min</sub> and N<sub>max</sub> are two asymptotes. <br />
<br>The parameter m is a curvature parameter. Larger m is, smaller is the curvature of the deceleration phase with the model. <br />
<br>The parameter n is a parameter related to the period lag. Larger n is, shorter is the period of lag. <br />
<br><br />
<br>N<sub>min</sub> is slightly lower than N<sub>0</sub>. When N is small at the initial state (N = N<sub>0</sub>) <i>i.e.</i> N is close to N<sub>min</sub>, N<sub>min</sub>/N is almost equal to 1. Therefore the term (1-(N<sub>min</sub>/N)) is nearly 0 and the growth is very slow. <br />
<br>If N decreases until it reaches N<sub>min</sub>, the term (1-(N<sub>min</sub>/N)) is equal to 0. Therefore the growth is null.<br />
<br> Similarly when N is equal to N<sub>max</sub>, the term (1-(N/N<sub>max</sub>)) is equal to 0 and the growth is blocked.</br><br />
<br />
To overcome this, we worked under two conditions: positive and negative growth. Theses conditions can be translated in two equations. This leads to the writing of this model:</p><br />
<center style="margin: 65px 0;"><img style="" src="https://static.igem.org/mediawiki/2014/f/f8/Form_part.png" alt="model"></center><br />
<br />
<br />
<br />
<br />
<p class="texte"><br />
with n = 1 and m = 0.5</br></br><br />
<br />
The term (1-(Nmin/N)) is not taken into account when there is growth. <br>The term (1-(N/Nmax)) is not taken into account when there is bacterial decay.</br><br />
Meteorological records of the Toulouse region during the years 2011-2013 are used to do average daily temperatures. Thus we can determine <i>B.subtilis</i> growth in a tree located in Toulouse during a year. This values are obtained for each day by the average on the highest and the lowest temperature.<br />
</br><br />
<br />
The density of green wood plane is about 650kg/m³. The average diameter of the trunks of the concerned trees is about 0.80m and 15m high. This represents a volume of 30m³. Therefore the weight of the trunk is 19.604kg.<br />
We need to add to this weight the weight of branches, twigs, about 25% of leaves and about 15% of roots (<a href="http://www.guichetdusavoir.org/viewtopic.php?t=25895">source-FR</a>).<br />
</br><br />
<!--pas compris ces deux dernières phrases--><br />
The average weight of a tree plane is 27,446kg. We inoculated 10mL of bacterial culture at 10⁹cfu/mL, <i>i.e.</i> 10^10 bacterial cells. This represents 3.64x10²cfu/g of fresh plant (N0).<br />
</p><br />
<br />
<center><img style="" src="https://static.igem.org/mediawiki/2014/5/53/Bacterial_growth.png" alt="Figure2"></center><br />
<p class="legend">Figure 2: (<span style="color:#000000; font-family:'Open Sans'; font-size:14px;">black</span>) <i>Bacillus subtilis</i> growth curve during one year (N is cell quantity by g of fresh plant). (<span style="color:#FF0040; font-family:'Open Sans'; font-size:14px;">red</span>) average temperature. (<span style="color:#0101DF; font-family:'Open Sans'; font-size:14px;">blue</span>) threshold at 10°C.</p><br />
<br />
<p class="texte"><br />
In our model, growth starts only from 10°C, which happens between March and April. This period seems to be suitable to put the strain in the tree. From December the temperature decreases below 4°C corresponding to the threshold below which bacteria die. <br />
</p><br />
<br />
<p class="title2"><br />
Discussion<br />
</p><br />
<br />
<p class="texte"><br />
In practice, temperature variations are certainly lower in trees than outside, especially if roots extend very deep. Composition of the tree sap must also intervene in the growth rate and nutrient content of sap is also temperature dependent. The effects of the decrease of the temperature in winter also induces a fall of the sap and this must also be involved in the disappearance of our strain in the tree. The period of <i>Bacillus subtilis</i> growth is certainly affected by the change of temperature, the rise of sap ans sap composition variations. All these parameters can consequently slow or fast the growth rate.<br />
<br />
The modeling work is done with the programming language 'R' script attached (See Annexe).<br />
</p><br />
<br />
<p class="title2"><br />
References<br />
</p><br />
<br />
<br />
<ul><br />
<li class="tree"><p class="texte"><br />
Xianling Ji, Guobing Lu, Yingping Gai, Chengchao Zheng & Zhimei Mu (2008) <b>Biological control against bacterial wilt and colonization of mulberry by an endophytic Bacillus subtilis strain.</b> FEMS Microbiol Ecol 65: 565–573<br />
</li></p><br />
<li class="tree"><p class="texte"><br />
A. Garnier(1977) <b>Transfert de sève brute dans le tronc des arbres aspects méthodologiques et physiologiques.</b> Ann. Sci. Foresi. 34 (1): 17-45<br />
</li></p><br />
<li class="tree"><p class="texte"><br />
Heikki Kallio , Tuija Teerinen , Seija Ahtonen , Meri Suihko , Reino R. Linko (1989) <b>Composition and properties of birch syrup (Betula pubescens).</b> J. Agric. Food Chem 37 (1): 51–54<br />
</li></p><br />
<li class="tree"><p class="texte"><br />
L. Rosso, J. R. Lobry & J. P. Flandrois (1992) AN <b>Unexpected Correlation between Cardinal Temperatures of Microbial Growth Highlighted by a New Model.</b> J. theor. Biol. 162 : 447-463<br />
</li></p><br />
<li class="tree"><p class="texte"><br />
Hiroshi Fujikawa (2010), <b>Development of a New Logistic Model for Microbial Growth in Foods.</b> Biocontrol of Science Vol 15: 75-80<br />
</li></p><br />
</ul><br />
<br />
<p class="title2"><br />
Annexe<br />
</p><br />
<br />
<p class="texte"> To upload the script and the table <a href="https://static.igem.org/mediawiki/2014/0/01/Annexes.zip">Click Here</a></p><br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/ModellingTeam:Toulouse/Modelling2014-10-17T21:39:06Z<p>Laureen: </p>
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<h2>Modeling</h2><br />
<p>To develop a predictive model</p><br />
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<p style="margin:0 auto; color:#696969; width:960px; padding-top:20px; font-size:16px;"> Project&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Modeling</p> <br />
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<br />
<p class="texte"><br />
<br />
Modeling is a tool used to simplify and study systems. It helps us to predict behavior thanks to bibliographic or experimental informations.</br><br />
<br />
The following modelisation focuses on the development of our engineered bacterium (called SubtiTree) in trees. The bacterial growth in trees seems to be unknown, thus we must infer <i>Bacillus subtilis'</i> behavior.</p><br />
<br />
<p class="title1"><br />
Bacterial Growth<br />
</p><br />
<br />
<p class="title2"><br />
Aim<br />
</p><br />
<br />
<p class="texte"><br />
<i>Bacillus subtilis</i> is a tree endophyte strain. A study showed that <i>Bacillus subtilis</i> could develop and fully colonize a tree, reaching a concentration of 10⁵ cells per gram of fresh plant. We need to know in which conditions the growth of <i>B. subtilis</i> is optimum in a tree and if the weather can stop its development during winter. Therefore we decided to work on the <i>Bacillus subtilis'</i> growth in function of the temperature during the year. <br />
<br>Modeling bacterial growth in a tree section generates some difficulties. We need to know the distance between two tree extremities (treetops and root) or the speed sap flow. However the speed sap flow can vary with temperature during the day and seasons cause of the type of sap (phloem, xylem). Furthermore a tree is not an homogeneous system: its roots, trunk and branches do not contain the same amount of sap and wood. <br>The average speed of the plane tree sap is 2.4m/h, which means that in a day the sap of a 30m tree will flow from one extremity to the other. Tree is reduced to a bioreactor.<br />
</p><br />
<p class="texte"><br />
We make the following hypothesis:</p><br />
<ol class="list1"><br />
<li><br />
According to the publication of <b>Xianling Ji</b> (See References), after six months of <i>Bacillus subtilis'</i> growth in a tree, bacteria cells reach a concentration of 10⁵ cells per gram of fresh plant. We assume that 10⁵ cells/g is the maximum concentration.<br />
</li><br />
<li><br />
The composition of the phloem is stable. There is no effect of depletion of the medium.<br />
</li><br />
<li><br />
Only temperature impacts on bacterial growth.<br />
</li><br />
<li><br />
It is believed that there is no leakage of cells.<br />
</li><br />
</ol><br />
<br />
<br />
<p class="title2"><br />
Method<br />
</p><br />
<br />
<p class="texte"><br />
An assessment of the <i>Bacillus subtilis</i> growth in a similar sap was performed in laboratory conditions with optimum growth medium for <i>Bacillus subtilis</i>. The composition sap used was the one from birch sap.<br><br />
In these conditions, the growth rate μ is optimal. From this value we can extrapolate a growth curve as a function of temperature. We used the <b>cardinal temperature model</b>: </p><br />
<br />
<center style="margin-bottom:50px;"><img style="" src="https://static.igem.org/mediawiki/2014/8/85/Formules_Rosso.png" alt="cardinal temperature model"></center><br />
<br />
<br />
<p class="texte"><br />
T: Temperature</br><br />
µ<sub>opt</sub>: Optimal growth rate</br><br />
µ: growth rate at temperature T</br><br />
T<sub>max</sub>: Maximum temperature supported by bacteria</br><br />
T<sub>min</sub>: Minimum temperature supported by bacteria</br><br />
T<sub>opt</sub>: Optimum temperature for the growth</br></br><br />
<br />
Necessary parameters for this function are minimun temperature T<sub>min</sub> and maximum temperature T<sub>max</sub>, optimal temperature for the growth T<sub>opt</sub> and optimal growth rate µ<sub>opt</sub>.</br><br />
</br><br />
T<sub>min</sub>: 10°C</br><br />
T<sub>max</sub>: 52°C</br><br />
T<sub>opt</sub>: 37°C</br><br />
µ<sub>opt</sub>: 8.5968 cfu/d</br></br><br />
<br />
The optimal growth rate (µ<sub>opt</sub>) is obtained experimentally with a similar birch sap environment.</br><br />
The growth rate is negative below 10°C (according to growth tests performed at 10°C and 4°C under similar conditions for the measurement of μ<sub>opt</sub>), survival rate after 24h was 0.3 % at 10°C and null at 4°C.<br><br />
Conditions apply:</p><br />
<br />
<p class="texte"> <br />
If<span style="color:#FFFFFF; font-family:'Open Sans'; font-size:14px;">__</span>| T<= 4°C -> µ = -1</br><br />
<span style="color:#FFFFFF; font-family:'Open Sans'; font-size:14px;">____</span>| 4°C<T<= 10°C -> µ = -0.97</br><br />
<span style="color:#FFFFFF; font-family:'Open Sans'; font-size:14px;">____</span>| T > 10°C -> µ = f(T) with f(T) egal to cardinal temperature model.</p><br />
<br />
<center style="margin-top: -52px;"><img style="" src="https://static.igem.org/mediawiki/2014/b/b1/Plot_growth_rate.png" alt="Figure1"></center><br />
<p class="legend">Figure 1: bacterial growth (µ) as a function of temperature</p><br />
<br />
<p class="texte"> A logistic model developed by <b>Hiroshi Fujikawa</b> (See References) is used to study bacterial growth.</p><br />
<br />
<p class="legend">General logistics formulas:</p><br />
<center style="margin:-44px 0 65px;"><img style="" src="https://static.igem.org/mediawiki/2014/c/c3/Form_general_fonction.png" alt="General logistics formulas"></center><br />
<br />
<br />
<p class="texte"><br />
In our case, the growth rate µ depends on the temperature. <br />
<br>N corresponds to the bacterial population, N<sub>min</sub> and N<sub>max</sub> are two asymptotes. <br />
<br>The parameter m is a curvature parameter. Larger m is, smaller is the curvature of the deceleration phase with the model. <br />
<br>The parameter n is a parameter related to the period lag. Larger n is, shorter is the period of lag. <br />
<br><br />
<br>N<sub>min</sub> is slightly lower than N<sub>0</sub>. When N is small at the initial state (N = N<sub>0</sub>) <i>i.e.</i> N is close to N<sub>min</sub>, N<sub>min</sub>/N is almost equal to 1. Therefore the term (1-(N<sub>min</sub>/N)) is nearly 0 and the growth is very slow. <br />
<br>If N decrease until reach N<sub>min</sub>, the term (1-(N<sub>min</sub>/N)) is equal to 0. Therefore the growth is null.<br />
<br> Similarly when N is equal to N<sub>max</sub> the term (1-(N/N<sub>max</sub>)) is equal to 0 and the growth is blocked.</br><br />
<br />
To overcome this we worked under two conditions: positive and negative growth. Theses conditions can be translated in two equations. This lead to the writing of this model:</p><br />
<center style="margin: 65px 0;"><img style="" src="https://static.igem.org/mediawiki/2014/f/f8/Form_part.png" alt="model"></center><br />
<br />
<br />
<br />
<br />
<p class="texte"><br />
with n = 1 and m = 0.5</br></br><br />
<br />
The term (1-(Nmin/N)) is not taken into account when there is growth. <br>The term (1-(N/Nmax)) is not taken into account when there is bacterial decay.</br><br />
Meteorological records of the Toulouse region during the years 2011-2013 are used to do average daily temperatures. Thus we can determine <i>B.subtilis</i> growth in a tree located in Toulouse during a year. This values are obtained for each day by the average on the highest and the lowest temperature.<br />
</br><br />
<br />
The density of green wood plane is about 650kg/m³. The average diameter of the trunks of the concerned trees is about 0.80m and 15m high. This represents a volume of 30m³. Therefore the weight of the trunk is 19,604kg.<br />
We need to add to this weight the weight of branches, twigs, about 25% of leaves and about 15% of roots (<a href="http://www.guichetdusavoir.org/viewtopic.php?t=25895">source-FR</a>).<br />
</br><br />
<!--pas compris ces deux dernières phrases--><br />
The average weight of a tree plane is 27,446kg. We inoculated 10mL of bacterial culture at 10⁹cfu/mL, <i>i.e.</i> 10^10 bacterial cells. This represents 3.64x10² cfu/g of fresh plant (N0).<br />
</p><br />
<br />
<center><img style="" src="https://static.igem.org/mediawiki/2014/5/53/Bacterial_growth.png" alt="Figure2"></center><br />
<p class="legend">Figure 2: (<span style="color:#000000; font-family:'Open Sans'; font-size:14px;">black</span>) <i>Bacillus subtilis</i> growth curve during one year (N is cell quantity by g of fresh plant). (<span style="color:#FF0040; font-family:'Open Sans'; font-size:14px;">red</span>) average temperature. (<span style="color:#0101DF; font-family:'Open Sans'; font-size:14px;">blue</span>) threshold at 10°C.</p><br />
<br />
<p class="texte"><br />
In our model, growth starts only from 10°C, which happens between March and April. This period seems to be suitable to put the strain in the tree. From December the temperature decreased below 4°C, threshold below which bacteria die. <br />
</p><br />
<br />
<p class="title2"><br />
Discussion<br />
</p><br />
<br />
<p class="texte"><br />
In practice, temperature variations are certainly lower in tree than outside, especially if roots extend very deep. Composition of the tree sap must also intervene in the growth rate and nutrient content of sap is also temperature dependent. The effects of the decrease of the temperature in winter also induces a fall of the sap and this must also be involved in the disappearance of our strain in the tree. The period of <i>Bacillus subtilis</i> growth is certainly affected by the change in temperature, the rise of sap ans sap composition variations. All these parameters can consequently slow or fast the growth rate.<br />
<br />
The modeling work is done with the programming language 'R' script attached (See Annexe).<br />
</p><br />
<br />
<p class="title2"><br />
References<br />
</p><br />
<br />
<br />
<ul><br />
<li class="tree"><p class="texte"><br />
Xianling Ji, Guobing Lu, Yingping Gai, Chengchao Zheng & Zhimei Mu (2008) <b>Biological control against bacterial wilt and colonization of mulberry by an endophytic Bacillus subtilis strain.</b> FEMS Microbiol Ecol 65: 565–573<br />
</li></p><br />
<li class="tree"><p class="texte"><br />
A. Garnier(1977) <b>Transfert de sève brute dans le tronc des arbres aspects méthodologiques et physiologiques.</b> Ann. Sci. Foresi. 34 (1): 17-45<br />
</li></p><br />
<li class="tree"><p class="texte"><br />
Heikki Kallio , Tuija Teerinen , Seija Ahtonen , Meri Suihko , Reino R. Linko (1989) <b>Composition and properties of birch syrup (Betula pubescens).</b> J. Agric. Food Chem 37 (1): 51–54<br />
</li></p><br />
<li class="tree"><p class="texte"><br />
L. Rosso, J. R. Lobry & J. P. Flandrois (1992) AN <b>Unexpected Correlation between Cardinal Temperatures of Microbial Growth Highlighted by a New Model.</b> J. theor. Biol. 162 : 447-463<br />
</li></p><br />
<li class="tree"><p class="texte"><br />
Hiroshi Fujikawa (2010), <b>Development of a New Logistic Model for Microbial Growth in Foods.</b> Biocontrol of Science Vol 15: 75-80<br />
</li></p><br />
</ul><br />
<br />
<p class="title2"><br />
Annexe<br />
</p><br />
<br />
<p class="texte"> To upload the script and the table <a href="https://static.igem.org/mediawiki/2014/0/01/Annexes.zip">Click Here</a></p><br />
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<p style="color:#696969; padding-top:20px; font-size:16px; float:left;"> Acknowlegdements&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Attributions</p> <br />
<ul class="topnav" id="topnav" style="top:15px;"><br />
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<h3 class="title2" style="margin-top:10px; color:#333;">Summary :</h3><br />
<ul class="menuleft"><br />
<li style="margin-top:25px;"><a href="#select1">Ulule</a></li><br />
<li><a href="#select2">Scientists</a></li><br />
<li><a href="#select3">Press</a></li><br />
<li><a href="#select4">Ethics</a></li><br />
<li><a href="#select5">Design and Communication</a></li><br />
<li><a href="#select6">Special thanks</a></li><br />
<li><a href="#select7">Premises</a></li><br />
<li><a href="#select8">Us</a></li><br />
</ul><br />
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<div class="column-right" style="width:75%; float:right;"><br />
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<p class="texte">Our team is proud to say that we almost did all the work by ourselves: finding sponsors, making communication tools about our project, finding the protocols to use and test them. However, we needed occasional assistance during our project, therefore we required help from different people: scientists, administrators, specialists and even lay persons. We would like to warmly thank all these persons for their contribution and their support. Our project could move forward because they offered their help.</p><br />
<br />
<p class="title1" id="select1">Ulule: a crowdfunding website to help us finding funds</p><br />
<br />
<table><br />
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<td><br />
<img src="https://static.igem.org/mediawiki/2014/2/23/LogoUlule.jpg" alt="Logo Ulule"><br />
</td><br />
<td valign="bottom"><p class="texte">Ulule is the first European crowdfunding website. We have posted our SubtiTree project on this site to collect funds for paying the Giant Jamboree fee. The collect was successful: we had 53 supporters and € 1,787collected!</br></br><br />
We warmly thank all the supporters, it was very encouraging to see that people have mobilized for us!<br />
</p><br />
</td><br />
</table><br />
</br><br />
<p class="texte">We would like to thank:</p><br />
<br />
<p class="nomsd">Debra Baker, François Barbeau, Valérie Chataigner, Brune Couffignal, Serge Couffignal, Jacques de Grenier, Guillaume Debar, Colas Decloitre, Caroline Delabre, Charlene Douard, Laurence Fournié, Rose Fournié, Chrystelle Gaucher, Nicolas Gilet, Aurélie Gosseau, Sarah Guiziou, Carole Jouve, Damien Kanitzer, Pierre Lagoutte, Anne-Marie Lang, Myriam Le Nours, Julie Mabille, Mylène Manière, Eddy Manière, Stéphanie Marty, Nathan Mirassou, Pierre Mirassou, Sophie Molina, Raymond Molina, Eric Molina, Ngoc Thu Hang Pham, Bruno Paci, Frédéric Paletou, Nicolas Perez, Jean Perron, Pierre Pettera, Lucas Picault, Nathalie Pineau, Elodie Prattico, Florence Rigaud, Marianne et Dominique Serres, Gilles Truan, Clémence Witzmann.</p><br />
<br />
<p class="title1" id="select2">Scientists<br />
</p><br />
<p class="title2">iGEM instructors<br />
</p><br />
<p class="texte">First of all, we warmly thank our amazing instructor: <b>Gilles Truan</b>. He was with us from the beginning to the end and even more! He is the master of cloning. He is the captain of PCR. Moreover, he knows how to grill sausages better than anyone!<br />
We kindly thank all the other instructors that helped us when we were lost with the scientific part or stressed by the lack of money. We especially thank <b>Brice Enjalbert</b>, <b>Florence Bordes</b>, <b>Kaymeuang Cam</b>, <b>Philippe Soucaille</b>, <b>Stéphane Guillouet</b>, <b>Isabelle Meynial-Salles</b> and <b>Alain Marty</b>. Your advice and encouragements were very precious to us, and you deserve these thanks for all the energy you spent answering our questions (and we know that we are very energy-consuming!).<br />
To finish, a special thanks to <b>Claude Maranges</b>, who helped us a lot with the sponsoring aspect and all the administrative problems that we encountered. Indeed, if you had an administrative issue, he is able to solve it in less than 2 minutes (5 minutes during the holidays)! He knows how to make everything all right in record time.<br />
</p><br />
<br />
<p class="title2"><i>Bacillus subtilis</i> specialists<br />
</p><br />
<p class="texte">Even if <i>Bacillus subtilis</i> is a gram-positive bacterium model, none of our instructors worked on it. It is why we asked other people's help when we had questions about <i>B. subtilis</i>.<br />
We would like to thank <b>Nathalie Campo</b> for her advice about <i>B. subtilis</i> culture, her encouragements and her kindness.<br />
Her friend <b>Thierry Doan</b> helped us too: he brilliantly answered our questions about <i>B. subtilis</i>.<br />
We also thank <b>Sarah Guiziou</b>, ex-iGEMer, for her advice and her protocols.<br />
</p><br />
<br />
<p class="title2">Fungi specialists<br />
</p><br />
<p class="texte"><br />
We are grateful to <b>Christophe Roux</b> who gave us two non-pathogenic strains of fungi: <i>Aspergillus brasiliensis</i> and <i>Chaetomium globusum</i>. We used them to test our SubtiTree system.<br />
</p><br />
<br />
<p class="title2">Other iGEM teams<br />
</p><br />
<p class="texte"><br />
Since several parts that we wanted to use were not in the iGEM kit plates, we asked iGEM teams to send us some parts:</br><br />
- Thanks to the <b>iGEM Warsaw team</b>, especially Radoslaw Stachowiak, for sending us the biobricks <br />
<a href="http://parts.igem.org/Part:BBa_K780000" target="_blank">BBa_K780000</a>, <a href="http://parts.igem.org/Part:BBa_K780001" target="_blank">BBa_K780001</a>, <a href="http://parts.igem.org/Part:BBa_K780002" target="_blank">BBa_K780002</a> and <a href="http://parts.igem.org/Part:BBa_K780003" target="_blank">BBa_K780003</a>.</br><br />
- Thanks to the <b>iGEM Utah State team</b>, especially Charles Miller, for sending us the biobrick <a href="http://parts.igem.org/Part:BBa_K1162001" target="_blank">BBa_K1162001</a>.</br><br />
- Thanks to the <b>iGEM Munich team</b>, especially Jara Radek, for sending us the biobrick <a href="http://parts.igem.org/Part:BBa_K823021" target="_blank">BBa_K823021</a>, <a href="http://parts.igem.org/Part:BBa_K823022" target="_blank">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K823023" target="_blank">BBa_K823023</a>.<br />
</p><br />
<br />
<p class="title2">Lab support<br />
</p><br />
<p class="texte"><br />
Thanks to <b>Sylvie Cancel</b> and <b>Yves Dutruy</b>, lab technicians, for helping us when we did not know where to find Yeast Extract or how to use the new refrigerated centrifuge. They were with us as soon as we began the manipulations and they answer to all our questions. </br><br />
A special thank to <b>Patrick Chekroun</b>, talended glass blower, for making the tubes system used in Chemotaxis tests.<br><br />
To finish, thanks to <b>Fabien Albert</b>, storeman of the LISBP, for his sympathy and his availability.<br />
</p><br />
<br />
<p class="title1" id="select3">Press<br />
</p><br />
<p class="texte">Since our project was very regional, a lot of different local newspapers were interested in SubtiTree.<br />
In this part, we would like to thank all the journalists that came to meet us:</br><br />
- <b>Hélène Ménal</b> for her article in the newspaper <br />
<a href="http://www.20minutes.fr/toulouse/1416215-20140708-guerisseurs-platanes" target="_blank">20 Minutes</a></br><br />
- <b>Philippe Font</b> for his article in the newspaper <br />
<a href="http://www.metronews.fr/toulouse/ils-etudient-une-bacterie-pour-soigner-les-platanes-du-canal-du-midi/mngj!FOMnxoEasL2s/" target="_blank">Metronews</a></br><br />
- <b>Bernard Davodeau</b> for his article in the newspaper <br />
<a href="http://www.ladepeche.fr/article/2014/07/15/1918689-un-espoir-de-traitement-pour-les-platanes-du-canal.html" target="_blank">La Dépêche</a></br><br />
- <b>Delphine Russeil</b> for her article in the newspaper La Voix du Midi</br><br />
- <b>Angélique Mangon</b> and <b>Jack Levé</b> for their television report on the channel <br />
<a href="http://france3-regions.francetvinfo.fr/midi-pyrenees/2014/07/09/des-etudiants-toulousains-developpent-une-bacterie-pour-sauver-les-platanes-514659.html" target="_blank">France 3 Midi-Pyrénées</a></br><br />
- <b>Philip Hemme</b> for his article about the French iGEM teams on the website <br />
<a href="http://labiotech.fr/les-equipes-francaise-lassaut-digem-2014-competition-mondiale-biologie-synthetique/" target="_blank">LaBiotech.fr</a></br><br />
- <b>Olivier Schlama</b> for his article in the newspaper Midi Libre</br><br />
- <b>Virginie Brancotte</b> for her article in the newspaper <br />
<a href="http://www.fluvialnet.com/murmures-actualites-des-etudiants-toulousains-inventent-une-methode-de-lutte-contre-le-chancre-colore/9576" target="_blank">Fluvial</a><br />
</p><br />
<br />
<p class="title1" id="select4">Ethics<br />
</p><br />
<p class="texte">We thank <b>Vincent Grégoire-Delory</b>, responsible of the ethics platform at Toulouse White Biotechnology, for coming to our lab and sharing his huge knowledge with us around pizzas and a good bottle of wine! He suggested us several points to consider and to deepen our ethical reflection.<br />
</p><br />
<br />
<p class="title1" id="select5">Design and Communication<br />
</p><br />
<p class="texte">We thank the <b>INSA Toulouse Communication Department</b> for giving us INSA Toulouse goodies and helping us with the organization of our acknowledgment day.<br><br />
Thanks to <b>Hélène Cabanac</b>, graphic designer, for her help with the design of our presentation and our poster.<br><br />
Thanks to the web designer <b>Adrien Nicod</b> for accepting to work with us and being patient with the remarks that each team member was saying!</br><br />
We are very proud of our logo and our wiki design, very professional but also modern and nice.<br />
<br />
</p><br />
<br />
<p class="title1" id="select6">Special thanks<br />
</p><br />
<p class="texte">We warmly thank <b>Didier Combes</b>, <b>Pierre Monsan</b> and <b>Nicolas Combébiac</b> for their support and their efficiency in the face of our administrative problems.</br><br />
A special thanks to <b>Gilbert Chauvel</b> for his interest into our project, his support, and his intervention with the Ministry.</br><br />
A special thanks to <b>Matthieu Arlat</b> for giving us a lot of good advice, helping us with the sponsoring part and his enthusiasm.</br><br />
To finish, thanks to the God of Synthetic Biology for giving us such results!<br />
</p><br />
<br />
<p class="title1" id="select7">Premises<br />
</p><br />
<p class="texte">We would like to thank the <b>INSA de Toulouse</b> for hosting us during the whole summer.<br />
</p><br />
<br />
<p class="title1" id="select8">Us<br />
</p><br />
<p class="texte">And finally, we thank... <b>us</b>. For being in the lab all the summer, for our devotion to the plane trees, for the perseverance in the bad days and for putting up each other.</br><br />
Therefore thanks to the entire team for this wonderful and exciting project!</br><br />
We would like to thank also our mums and dads because without them we would not be there and thus the project would not have been done! And also the mum and dad of our mums and dads and the mum and dad of the mum and dad of our mums and dads and... you should get it!!<br />
</p><br />
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<h3 class="title2" style="margin-top:10px; color:#333;">Summary :</h3><br />
<ul class="menuleft"><br />
<li style="margin-top:25px;"><a href="#select1">Ulule</a></li><br />
<li><a href="#select2">Scientists</a></li><br />
<li><a href="#select3">Press</a></li><br />
<li><a href="#select4">Ethics</a></li><br />
<li><a href="#select5">Design and Communication</a></li><br />
<li><a href="#select6">Special thanks</a></li><br />
<li><a href="#select7">Premises</a></li><br />
<li><a href="#select8">Us</a></li><br />
</ul><br />
</div><br />
<br />
<div class="column-right" style="width:75%; float:right;"><br />
<br />
<p class="texte">Our team is proud to say that we almost did all the work by ourselves: finding sponsors, making communication tools about our project, finding the protocols to use and test them. However, we needed occasional assistance during our project, therefore we required help from different people: scientists, administrators, specialists and even lay persons. We would like to warmly thank all these persons for their contribution and their support. Our project could move forward because they offered their help.</p><br />
<br />
<p class="title1" id="select1">Ulule: a crowdfunding website to help us finding funds</p><br />
<br />
<table><br />
<tr><br />
<td><br />
<img src="https://static.igem.org/mediawiki/2014/2/23/LogoUlule.jpg" alt="Logo Ulule"><br />
</td><br />
<td valign="bottom"><p class="texte">Ulule is the first European crowdfunding website. We have posted our SubtiTree project on this site to collect funds for paying the Giant Jamboree fee. The collect was successful: we had 53 supporters and € 1,787collected!</br></br><br />
We warmly thank all the supporters, it was very encouraging to see that people have mobilized for us!<br />
</p><br />
</td><br />
</table><br />
</br><br />
<p class="texte">We would like to thank:</p><br />
<br />
<p class="nomsd">Debra Baker, François Barbeau, Valérie Chataigner, Brune Couffignal, Serge Couffignal, Jacques de Grenier, Guillaume Debar, Colas Decloitre, Caroline Delabre, Charlene Douard, Laurence Fournié, Rose Fournié, Chrystelle Gaucher, Nicolas Gilet, Aurélie Gosseau, Sarah Guiziou, Carole Jouve, Damien Kanitzer, Pierre Lagoutte, Anne-Marie Lang, Myriam Le Nours, Julie Mabille, Mylène Manière, Eddy Manière, Stéphanie Marty, Nathan Mirassou, Pierre Mirassou, Sophie Molina, Raymond Molina, Eric Molina, Ngoc Thu Hang Pham, Bruno Paci, Frédéric Paletou, Nicolas Perez, Jean Perron, Pierre Pettera, Lucas Picault, Nathalie Pineau, Elodie Prattico, Florence Rigaud, Marianne et Dominique Serres, Gilles Truan, Clémence Witzmann.</p><br />
<br />
<p class="title1" id="select2">Scientists<br />
</p><br />
<p class="title2">iGEM instructors<br />
</p><br />
<p class="texte">First of all, we warmly thank our amazing instructor: <b>Gilles Truan</b>. He was with us from the beginning to the end and even more! He is the master of cloning. He is the captain of PCR. Moreover, he knows how to grill sausages better than anyone!<br />
We kindly thank all the other instructors that helped us when we were lost with the scientific part or stressed by the lack of money. We especially thank <b>Brice Enjalbert</b>, <b>Florence Bordes</b>, <b>Kaymeuang Cam</b>, <b>Philippe Soucaille</b>, <b>Stéphane Guillouet</b>, <b>Isabelle Meynial-Salles</b> and <b>Alain Marty</b>. Your advice and encouragements were very precious to us, and you deserve these thanks for all the energy you spent answering our questions (and we know that we are very energy-consuming!).<br />
To finish, a special thanks to <b>Claude Maranges</b>, who helped us a lot with the sponsoring aspect and all the administrative problems that we encountered. Indeed, if you had an administrative issue, he is able to solve it in less than 2 minutes (5 minutes during the holidays)! He knows how to make everything all right in record time.<br />
</p><br />
<br />
<p class="title2"><i>Bacillus subtilis</i> specialists<br />
</p><br />
<p class="texte">Even if <i>Bacillus subtilis</i> is a gram-positive bacterium model, none of our instructors worked on it. It is why we asked other people's help when we had questions about B. subtilis.<br />
We would like to thank <b>Nathalie Campo</b> for her advice about <i>B. subtilis</i> culture, her encouragements and her kindness.<br />
Her friend <b>Thierry Doan</b> helped us too: he brilliantly answered our questions about <i>B. subtilis</i>.<br />
We also thank <b>Sarah Guiziou</b>, ex-iGEMer, for her advice and her protocols<br />
</p><br />
<br />
<p class="title2">Fungi specialists<br />
</p><br />
<p class="texte"><br />
We are grateful to <b>Christophe Roux</b> who gave us two non-pathogenic strains of fungi: <i>Aspergillus brasiliensis</i> and <i>Chaetomium globusum</i>. We used them to test our SubtiTree system.<br />
</p><br />
<br />
<p class="title2">Other iGEM teams<br />
</p><br />
<p class="texte"><br />
Since several parts that we wanted to use were not in the iGEM kit plates, we asked iGEM teams to send us some parts:</br><br />
- Thanks to the <b>iGEM Warsaw team</b>, especially Radoslaw Stachowiak, for sending us the biobricks <br />
<a href="http://parts.igem.org/Part:BBa_K780000" target="_blank">BBa_K780000</a>, <a href="http://parts.igem.org/Part:BBa_K780001" target="_blank">BBa_K780001</a>, <a href="http://parts.igem.org/Part:BBa_K780002" target="_blank">BBa_K780002</a> and <a href="http://parts.igem.org/Part:BBa_K780003" target="_blank">BBa_K780003</a>.</br><br />
- Thanks to the <b>iGEM Utah State team</b>, especially Charles Miller, for sending us the biobrick <a href="http://parts.igem.org/Part:BBa_K1162001" target="_blank">BBa_K1162001</a>.</br><br />
- Thanks to the <b>iGEM Munich team</b>, especially Jara Radek, for sending us the biobrick <a href="http://parts.igem.org/Part:BBa_K823021" target="_blank">BBa_K823021</a>, <a href="http://parts.igem.org/Part:BBa_K823022" target="_blank">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K823023" target="_blank">BBa_K823023</a>.<br />
</p><br />
<br />
<p class="title2">Lab support<br />
</p><br />
<p class="texte"><br />
Thanks to <b>Sylvie Cancel</b> and <b>Yves Dutruy</b>, lab technicians, for helping us when we did not know where to find Yeast Extract or how to use the new refrigerated centrifuge. They were with us as soon as we began the manipulations and they answer to all our questions. </br><br />
A special thank to <b>Patrick Chekroun</b>, talended glass blower, for making the tubes system used in Chemotaxis tests.<br><br />
To finish, thanks to <b>Fabien Albert</b>, storeman of the LISBP, for his sympathy and his availability.<br />
</p><br />
<br />
<p class="title1" id="select3">Press<br />
</p><br />
<p class="texte">Since our project was very regional, a lot of different local newspapers were interested in SubtiTree.<br />
In this part, we would like to thank all the journalists that came to meet us:</br><br />
- <b>Hélène Ménal</b> for her article in the newspaper <br />
<a href="http://www.20minutes.fr/toulouse/1416215-20140708-guerisseurs-platanes" target="_blank">20 Minutes</a></br><br />
- <b>Philippe Font</b> for his article in the newspaper <br />
<a href="http://www.metronews.fr/toulouse/ils-etudient-une-bacterie-pour-soigner-les-platanes-du-canal-du-midi/mngj!FOMnxoEasL2s/" target="_blank">Metronews</a></br><br />
- <b>Bernard Davodeau</b> for his article in the newspaper <br />
<a href="http://www.ladepeche.fr/article/2014/07/15/1918689-un-espoir-de-traitement-pour-les-platanes-du-canal.html" target="_blank">La Dépêche</a></br><br />
- <b>Delphine Russeil</b> for her article in the newspaper La Voix du Midi</br><br />
- <b>Angélique Mangon</b> and <b>Jack Levé</b> for their television report on the channel <br />
<a href="http://france3-regions.francetvinfo.fr/midi-pyrenees/2014/07/09/des-etudiants-toulousains-developpent-une-bacterie-pour-sauver-les-platanes-514659.html" target="_blank">France 3 Midi-Pyrénées</a></br><br />
- <b>Philip Hemme</b> for his article about the French iGEM teams on the website <br />
<a href="http://labiotech.fr/les-equipes-francaise-lassaut-digem-2014-competition-mondiale-biologie-synthetique/" target="_blank">LaBiotech.fr</a></br><br />
- <b>Olivier Schlama</b> for his article in the newspaper Midi Libre</br><br />
- <b>Virginie Brancotte</b> for her article in the newspaper <br />
<a href="http://www.fluvialnet.com/murmures-actualites-des-etudiants-toulousains-inventent-une-methode-de-lutte-contre-le-chancre-colore/9576" target="_blank">Fluvial</a><br />
</p><br />
<br />
<p class="title1" id="select4">Ethics<br />
</p><br />
<p class="texte">We thank <b>Vincent Grégoire-Delory</b>, responsible of the ethics platform at Toulouse White Biotechnology, for coming to our lab and sharing his huge knowledge with us around pizzas and a good bottle of wine! He suggested us several points to consider and to deepen our ethical reflection.<br />
</p><br />
<br />
<p class="title1" id="select5">Design and Communication<br />
</p><br />
<p class="texte">We thank the <b>INSA Toulouse Communication Department</b> for giving us INSA Toulouse goodies and helping us with the organization of our acknowledgment day.<br><br />
Thanks to <b>Hélène Cabanac</b>, graphic designer, for her help with the design of our presentation and our poster.<br><br />
Thanks to the web designer <b>Adrien Nicod</b> for accepting to work with us and being patient with the remarks that each team member was saying!</br><br />
We are very proud of our logo and our wiki design, very professional but also modern and nice.<br />
<br />
</p><br />
<br />
<p class="title1" id="select6">Special thanks<br />
</p><br />
<p class="texte">We warmly thank <b>Didier Combes</b>, <b>Pierre Monsan</b> and <b>Nicolas Combébiac</b> for their support and their efficiency in the face of our administrative problems.</br><br />
A special thanks to <b>Gilbert Chauvel</b> for his interest into our project, his support, and his intervention with the Ministry.</br><br />
A special thanks to <b>Matthieu Arlat</b> for giving us a lot of good advice, helping us with the sponsoring part and his enthusiasm.</br><br />
To finish, thanks to the God of Synthetic Biology for giving us such results!<br />
</p><br />
<br />
<p class="title1" id="select7">Premises<br />
</p><br />
<p class="texte">We would like to thank the <b>INSA de Toulouse</b> for hosting us during the whole summer.<br />
</p><br />
<br />
<p class="title1" id="select8">Us<br />
</p><br />
<p class="texte">And finally, we thank... <b>us</b>. For being in the lab all the summer, for our devotion to the plane trees, for the perseverance in the bad days and for putting up each other.</br><br />
Therefore thanks to the entire team for this wonderful and exciting project!</br><br />
We would like to thank also our mums and dads because without them we would not be there and thus the project would not have been done! And also the mum and dad of our mums and dads and the mum and dad of the mum and dad of our mums and dads and... you should get it!!<br />
</p><br />
<br />
</div><br />
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<!-------------------------------- FOOTER ---------------------------------><br />
{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/Acknowledgements/AttributionsTeam:Toulouse/Acknowledgements/Attributions2014-10-16T20:28:03Z<p>Laureen: </p>
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<p style="color:#696969; padding-top:20px; font-size:16px; float:left;"> Acknowlegdements&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Attributions</p> <br />
<ul class="topnav" id="topnav" style="top:15px;"><br />
<br />
</ul><br />
</div><br />
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<br />
<div id="column-left"><br />
<h3 class="title2" style="margin-top:10px; color:#333;">Summary :</h3><br />
<ul class="menuleft"><br />
<li style="margin-top:25px;"><a href="#select1">Ulule</a></li><br />
<li><a href="#select2">Scientists</a></li><br />
<li><a href="#select3">Press</a></li><br />
<li><a href="#select4">Ethics</a></li><br />
<li><a href="#select5">Design and Communication</a></li><br />
<li><a href="#select6">Special thanks</a></li><br />
<li><a href="#select7">Premises</a></li><br />
<li><a href="#select8">Us</a></li><br />
</ul><br />
</div><br />
<br />
<div class="column-right" style="width:75%; float:right;"><br />
<br />
<p class="texte">Our team is proud to say that we almost did all the work by ourselves: finding sponsors, making communication tools about our project, finding the protocols to use and test them. However, we needed occasional assistance during our project, therefore we required help from different people: scientists, administrators, specialists and even lay persons. We would like to warmly thank all these persons for their contribution and their support. Our project could move forward because they offered their help.</p><br />
<br />
<p class="title1" id="select1">Ulule: a crowdfunding website to help us finding funds</p><br />
<br />
<table><br />
<tr><br />
<td><br />
<img src="https://static.igem.org/mediawiki/2014/2/23/LogoUlule.jpg" alt="Logo Ulule"><br />
</td><br />
<td valign="bottom"><p class="texte">Ulule is the first European crowdfunding website. We have posted our SubtiTree project on this site to collect funds for paying the Giant Jamboree fee. The collect was successful: we had 53 supporters and 1,787€ collected!</br></br><br />
We warmly thank all the supporters, it was very encouraging to see that people have mobilized for us!<br />
</p><br />
</td><br />
</table><br />
</br><br />
<p class="texte">We would like to thank:</p><br />
<br />
<p class="nomsd">Debra Baker, François Barbeau, Valérie Chataigner, Brune Couffignal, Serge Couffignal, Jacques de Grenier, Guillaume Debar, Colas Decloitre, Caroline Delabre, Charlene Douard, Laurence Fournié, Rose Fournié, Chrystelle Gaucher, Nicolas Gilet, Aurélie Gosseau, Sarah Guiziou, Carole Jouve, Damien Kanitzer, Pierre Lagoutte, Anne-Marie Lang, Myriam Le Nours, Julie Mabille, Mylène Manière, Eddy Manière, Stéphanie Marty, Nathan Mirassou, Pierre Mirassou, Sophie Molina, Raymond Molina, Eric Molina, Ngoc Thu Hang Pham, Bruno Paci, Frédéric Paletou, Nicolas Perez, Jean Perron, Pierre Pettera, Lucas Picault, Nathalie Pineau, Elodie Prattico, Florence Rigaud, Marianne et Dominique Serres, Gilles Truan, Clémence Witzmann.</p><br />
<br />
<p class="title1" id="select2">Scientists<br />
</p><br />
<p class="title2">iGEM instructors<br />
</p><br />
<p class="texte">First of all, we warmly thank our amazing instructor: <b>Gilles Truan</b>. He was with us from the beginning to the end and even more! He is the master of cloning. He is the captain of PCR. Moreover, he knows how to grill sausages better than anyone!<br />
We kindly thank all the other instructors that helped us when we were lost with the scientific part or stressed by the lack of money. We especially thank <b>Brice Enjalbert</b>, <b>Florence Bordes</b>, <b>Kaymeuang Cam</b>, <b>Philippe Soucaille</b>, <b>Stéphane Guillouet</b>, <b>Isabelle Meynial-Salles</b> and <b>Alain Marty</b>. Your advice and encouragements were very precious to us, and you deserve these thanks for all the energy you spent answering our questions (and we know that we are very energy-consuming!).<br />
To finish, a special thanks to <b>Claude Maranges</b>, who helped us a lot with the sponsoring aspect and all the administrative problems that we encountered. Indeed, if you had an administrative issue, he is able to solve it in less than 2 minutes (5 minutes during the holidays)! He knows how to make everything all right in record time.<br />
</p><br />
<br />
<p class="title2"><i>Bacillus subtilis</i> specialists<br />
</p><br />
<p class="texte">Even if <i>Bacillus subtilis</i> is a gram-positive bacterium model, none of our instructors worked on it. It is why we asked other people's help when we had questions about B. subtilis.<br />
We would like to thank <b>Nathalie Campo</b> for her advice about <i>B. subtilis</i> culture, her encouragements and her kindness.<br />
Her friend <b>Thierry Doan</b> helped us too: he brilliantly answered our questions about <i>B. subtilis</i>.<br />
We also thank <b>Sarah Guiziou</b>, ex-iGEMer, for her advice and her protocols<br />
</p><br />
<br />
<p class="title2">Fungi specialists<br />
</p><br />
<p class="texte"><br />
We are grateful to <b>Christophe Roux</b> who gave us two non-pathogenic strains of fungi: <i>Aspergillus brasiliensis</i> and <i>Chaetomium globusum</i>. We used them to test our SubtiTree system.<br />
</p><br />
<br />
<p class="title2">Other iGEM teams<br />
</p><br />
<p class="texte"><br />
Since several parts that we wanted to use were not in the iGEM kit plates, we asked iGEM teams to send us some parts:</br><br />
- Thanks to the <b>iGEM Warsaw team</b>, especially Radoslaw Stachowiak, for sending us the biobricks <br />
<a href="http://parts.igem.org/Part:BBa_K780000" target="_blank">BBa_K780000</a>, <a href="http://parts.igem.org/Part:BBa_K780001" target="_blank">BBa_K780001</a>, <a href="http://parts.igem.org/Part:BBa_K780002" target="_blank">BBa_K780002</a> and <a href="http://parts.igem.org/Part:BBa_K780003" target="_blank">BBa_K780003</a>.</br><br />
- Thanks to the <b>iGEM Utah State team</b>, especially Charles Miller, for sending us the biobrick <a href="http://parts.igem.org/Part:BBa_K1162001" target="_blank">BBa_K1162001</a>.</br><br />
- Thanks to the <b>iGEM Munich team</b>, especially Jara Radek, for sending us the biobrick <a href="http://parts.igem.org/Part:BBa_K823021" target="_blank">BBa_K823021</a>, <a href="http://parts.igem.org/Part:BBa_K823022" target="_blank">BBa_K823022</a> and <a href="http://parts.igem.org/Part:BBa_K823023" target="_blank">BBa_K823023</a>.<br />
</p><br />
<br />
<p class="title2">Lab support<br />
</p><br />
<p class="texte"><br />
Thanks to <b>Sylvie Cancel</b> and <b>Yves Dutruy</b>, lab technicians, for helping us when we did not know where to find Yeast Extract or how to use the new refrigerated centrifuge. They were with us as soon as we began the manipulations and they answer to all our questions. </br><br />
A special thank to <b>Patrick Chekroun</b>, talended glass blower, for making the tubes system used in Chemotaxis tests.<br><br />
To finish, thanks to <b>Fabien Albert</b>, storeman of the LISBP, for his sympathy and his availability.<br />
</p><br />
<br />
<p class="title1" id="select3">Press<br />
</p><br />
<p class="texte">Since our project was very regional, a lot of different local newspapers were interested in SubtiTree.<br />
In this part, we would like to thank all the journalists that came to meet us:</br><br />
- <b>Hélène Ménal</b> for her article in the newspaper <br />
<a href="http://www.20minutes.fr/toulouse/1416215-20140708-guerisseurs-platanes" target="_blank">20 Minutes</a></br><br />
- <b>Philippe Font</b> for his article in the newspaper <br />
<a href="http://www.metronews.fr/toulouse/ils-etudient-une-bacterie-pour-soigner-les-platanes-du-canal-du-midi/mngj!FOMnxoEasL2s/" target="_blank">Metronews</a></br><br />
- <b>Bernard Davodeau</b> for his article in the newspaper <br />
<a href="http://www.ladepeche.fr/article/2014/07/15/1918689-un-espoir-de-traitement-pour-les-platanes-du-canal.html" target="_blank">La Dépêche</a></br><br />
- <b>Delphine Russeil</b> for her article in the newspaper La Voix du Midi</br><br />
- <b>Angélique Mangon</b> and <b>Jack Levé</b> for their television report on the channel <br />
<a href="http://france3-regions.francetvinfo.fr/midi-pyrenees/2014/07/09/des-etudiants-toulousains-developpent-une-bacterie-pour-sauver-les-platanes-514659.html" target="_blank">France 3 Midi-Pyrénées</a></br><br />
- <b>Philip Hemme</b> for his article about the French iGEM teams on the website <br />
<a href="http://labiotech.fr/les-equipes-francaise-lassaut-digem-2014-competition-mondiale-biologie-synthetique/" target="_blank">LaBiotech.fr</a></br><br />
- <b>Olivier Schlama</b> for his article in the newspaper Midi Libre</br><br />
- <b>Virginie Brancotte</b> for her article in the newspaper <br />
<a href="http://www.fluvialnet.com/murmures-actualites-des-etudiants-toulousains-inventent-une-methode-de-lutte-contre-le-chancre-colore/9576" target="_blank">Fluvial</a><br />
</p><br />
<br />
<p class="title1" id="select4">Ethics<br />
</p><br />
<p class="texte">We thank <b>Vincent Grégoire-Delory</b>, responsible of the ethics platform at Toulouse White Biotechnology, for coming to our lab and sharing his huge knowledge with us around pizzas and a good bottle of wine! He suggested us several points to consider and to deepen our ethical reflection.<br />
</p><br />
<br />
<p class="title1" id="select5">Design and Communication<br />
</p><br />
<p class="texte">We thank the <b>INSA Toulouse Communication Department</b> for giving us INSA Toulouse goodies and helping us with the organization of our acknowledgment day.<br><br />
Thanks to <b>Hélène Cabanac</b>, graphic designer, for her help with the design of our presentation and our poster.<br><br />
Thanks to the web designer <b>Adrien Nicod</b> for accepting to work with us and being patient with the remarks that each team member was saying!</br><br />
We are very proud of our logo and our wiki design, very professional but also modern and nice.<br />
<br />
</p><br />
<br />
<p class="title1" id="select6">Special thanks<br />
</p><br />
<p class="texte">We warmly thank <b>Didier Combes</b>, <b>Pierre Monsan</b> and <b>Nicolas Combébiac</b> for their support and their efficiency in the face of our administrative problems.</br><br />
A special thanks to <b>Gilbert Chauvel</b> for his interest into our project, his support, and his intervention with the Ministry.</br><br />
A special thanks to <b>Matthieu Arlat</b> for giving us a lot of good advice, helping us with the sponsoring part and his enthusiasm.</br><br />
To finish, thanks to the God of Synthetic Biology for giving us such results!<br />
</p><br />
<br />
<p class="title1" id="select7">Premises<br />
</p><br />
<p class="texte">We would like to thank the <b>INSA de Toulouse</b> for hosting us during the whole summer.<br />
</p><br />
<br />
<p class="title1" id="select8">Us<br />
</p><br />
<p class="texte">And finally, we thank... <b>us</b>. For being in the lab all the summer, for our devotion to the plane trees, for the perseverance in the bad days and for putting up each other.</br><br />
Therefore thanks to the entire team for this wonderful and exciting project!</br><br />
We would like to thank also our mums and dads because without them we would not be there and thus the project would not have been done! And also the mum and dad of our mums and dads and the mum and dad of the mum and dad of our mums and dads and... you should get it!!<br />
</p><br />
<br />
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<br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/Project/project-contextTeam:Toulouse/Project/project-context2014-10-16T20:18:14Z<p>Laureen: </p>
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<div class="banniere-content"><br />
<h2>Project context</h2><br />
<p>A threatened heritage</p><br />
</div><br />
</div><br />
</div><br />
<br />
<div class="fils-ariane" style="width:100%; height:60px; background:#ededed;"><br />
<p style="margin:0 auto; color:#696969; width:960px; padding-top:20px; font-size:16px;"> Project&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Context project</p> <br />
</div><br />
<br />
<br />
<div id="innercontenthome"><br />
<div class="centering" style="padding-top: 85px; padding-bottom:40px;"><br />
<br />
<!--Short description : à changer!!!--><br />
<br />
<p class="texte">One of Europe Wonders is without a doubt the Toulouse Canal du Midi. Situated in the heart of our city and going through Southern France, our lab is only 5 minutes walk from the Canal du Midi. Everyone in the team could enjoy the quietness and loveliness of this charming place. Unfortunately, this heritage is threatened: a phytopathogenic fungus, <i>Ceratocystis platani</i>, is devastating the trees lining the Canal.<br />
</p><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2014/1/14/Overview_context.jpg"/></center><br />
<br></br><br />
<br />
<p class="texte">The situation is alarming: today the only solution is preventive tree cutting to stem the fungal epidemic. The consequences are disastrous. Not only for the beauty of the landscapes but it has also huge environmental, social and economic costs. Today, 42,000 plane trees are threatened, knowing that cutting and planting a new resistant tree is about €4,000.<br />
</p><br />
<br />
<p class="title1">The Canal du midi</p> <br />
<p class="texte">This ingenious masterpiece respects the environment in which it is harmoniously integrated since 1681 by Pierre-Paul Riquet.<br />
Characterized as a summit level canal, it culminates at 189 meters of altitude and permits the achievement of water through the sea.<br />
In December 7th 1996, the UNESCO registers Le Canal du Midi on the list of world’s legacy. The first plane trees bordering the channel appeared lately after its creation in 1776. Resolving the erosion of riverbanks and the evaporation of the water, the plane imposes itself as a dominant and emblematic marrow. The wooded legacy is estimated to about 42,000 trees, it contributes to the landscape and aesthetic.<br />
Today, the canal du midi is primarily used for tourism, recreation and housing. Busier than the Seine (Paris river), it accounts for one fifth of French river tourism and 80 % tourists are foreigners. Boaters seeking for tranquility, quietness and a unique environment mainly navigate over it.<br />
</p><br />
<br />
<p class="title1">Canker Stain</p> <br />
<div style="float:left; width:215px; margin-right:40px"><img style="width:200px; float:left;" src="https://static.igem.org/mediawiki/2014/5/5e/Ceratocystis.jpg"></div><br />
<p class="texte">The canker stain is a disease caused by <i>Ceratocystis platani</i>, a microscopic fungus that exclusively attacks Plane Trees.<br />
Probably introduced in France by infected wooden munitions cases coming from United States of America in 1945, the fungus introduces itself inside the sain tree’s heart, blocks the sap flow and kills it in only 2 to 5 years.<br />
On the regulatory side, on July 31th 2000 a ministerial order has classed <I>Ceratocystis platani</I> as an harmful organism for plants. This order leads to a mandatory fight against the fungus.<br />
</p><br />
<br />
<br></br><br />
<br />
<p class="title1">Fighting the plane trees pathogen</p> <br />
<p class="texte">Regarding this imminent threat, some solutions emerged.<br />
</br>First of all, chemical fungicides were used. However, the community realized it was impossible to eradicate the whole fungus. <br />
</br>Therefore, the French Institut National de Recherche Agronomique (INRA) created a new type of plane trees named Platanor which were resistant to the fungus infection. Unfortunately, this extreme measure could not save the already contaminated trees nearby the Canal du Midi.<br />
</br>Today, the only option remains in the tree-cutting of the contaminated plane trees. Nevertheless, this previous solution is not optimal because of the high cost and the major ecological, touristic and social impact for our heritage.<br />
</p><br />
<br />
<br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/Project/project-contextTeam:Toulouse/Project/project-context2014-10-16T20:13:21Z<p>Laureen: </p>
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<p>A threatened heritage</p><br />
</div><br />
</div><br />
</div><br />
<br />
<div class="fils-ariane" style="width:100%; height:60px; background:#ededed;"><br />
<p style="margin:0 auto; color:#696969; width:960px; padding-top:20px; font-size:16px;"> Project&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Context project</p> <br />
</div><br />
<br />
<br />
<div id="innercontenthome"><br />
<div class="centering" style="padding-top: 85px; padding-bottom:40px;"><br />
<br />
<!--Short description : à changer!!!--><br />
<br />
<p class="texte">One of Europe Wonders is without a doubt the Toulouse Canal du Midi. Situated in the heart of our city and going through Southern France, our lab is only 5 minutes walk from the Canal du Midi. Everyone in the team could enjoy the quietness and loveliness of this charming place. Unfortunately, this heritage is threatened: a phytopathogenic fungus, <i>Ceratocystis platani</i>, is devastating the trees lining the Canal.<br />
</p><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2014/1/14/Overview_context.jpg"/></center><br />
<br></br><br />
<br />
<p class="texte">The situation is alarming: today the only solution is preventive tree cutting to stem the fungal epidemic. The consequences are disastrous. Not only for the beauty of the landscapes but it has also huge environmental, social and economic costs. Today, 42,000 plane trees are threatened, knowing that cutting and planting a new resistant tree is about €4,000.<br />
</p><br />
<br />
<p class="title1">The Canal du midi</p> <br />
<p class="texte">This ingenious masterpiece respects the environment in which it is harmoniously integrated since 1681 by Pierre-Paul Riquet.<br />
Characterized as a summit level canal, it culminates at 189 meters of altitude and permits the achievement of water through the sea.<br />
In December 7th 1996, the UNESCO registers Le Canal du Midi on the list of world’s legacy. The first plane trees bordering the channel appeared lately after its creation in 1776. Resolving the erosion of riverbanks and the evaporation of the water, the plane imposes itself as a dominant and emblematic marrow. The wooded legacy is estimated to about 42,000 trees, it contributes to the landscape and aesthetic.<br />
Today, the canal du midi is primarily used for tourism, recreation and housing. Busier than the Seine (Paris river), it accounts for one fifth of French river tourism and 80 % tourists are foreigners. Boaters seeking for tranquility, quietness and a unique environment mainly navigate over it.<br />
</p><br />
<br />
<p class="title1">Canker Stain</p> <br />
<div style="float:left; width:215px; margin-right:40px"><img style="width:200px; float:left;" src="https://static.igem.org/mediawiki/2014/5/5e/Ceratocystis.jpg"></div><br />
<p class="texte">The canker stain is a disease caused by <i>Ceratocystis platani</i>, a microscopic fungus that exclusively attacks Plane Trees.<br />
Probably introduced in France by infected wooden munitions cases coming from United States of America in 1945, the fungus introduces itself inside the sain tree’s heart, blocks the sap flow and kills it in only 2 to 5 years.<br />
On the regulatory side, the July 31th 2000 modified ministerial order classes <i>Ceratocystis</i> as a harmful organism for plants. This order leads to a mandatory fight against the fungus.<br />
</p><br />
<br />
<br></br><br />
<br />
<p class="title1">Fighting the plane trees pathogen</p> <br />
<p class="texte">Regarding this imminent threat, some solutions emerged.<br />
</br>First of all, chemical fungicides were used. However, the community realized it was impossible to eradicate the whole fungus. <br />
</br>Therefore, the French Institut National de Recherche Agronomique (INRA) created a new type of plane trees named Platanor which were resistant to the fungus infection. Unfortunately, this extreme measure could not save the already contaminated trees nearby the Canal du Midi.<br />
</br>Today, the only option remains in the tree-cutting of the contaminated plane trees. Nevertheless, this previous solution is not optimal because of the high cost and the major ecological, touristic and social impact for our heritage.<br />
</p><br />
<br />
<br />
<!-- Navigation section --> <br />
<br />
<div class="page-nav" style="border-top:1px solid #cccccc; padding-top:40px; margin-top:40px;"><br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/Project/project-contextTeam:Toulouse/Project/project-context2014-10-16T20:11:18Z<p>Laureen: </p>
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<p class="texte">One of Europe Wonders is without a doubt the Toulouse Canal du Midi. Situated in the heart of our city and going through Southern France, our lab is only 5 minutes walk from the Canal du Midi. Everyone in the team could enjoy the quietness and loveliness of this charming place. Unfortunately, this heritage is threatened: a phytopathogenic fungus, <i>Ceratocystis platani</i>, is devastating the trees lining the Canal.<br />
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<p class="texte">The situation is alarming: today the only solution is preventive tree cutting to stem the fungal epidemic. The consequences are disastrous. Not only for the beauty of the landscapes but it has also huge environmental, social and economic costs. Today, 42,000 plane trees are threatened, knowing that cutting and planting a new resistant tree is about €4,000.<br />
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<p class="title1">The Canal du midi</p> <br />
<p class="texte">This ingenious masterpiece respects the environment in which it is harmoniously integrated since 1681 by Pierre-Paul Riquet.<br />
Characterized as a summit level canal, it culminates at 189 meters of altitude and permits the achievement of water through the sea.<br />
In December 7th, 1996, the UNESCO registers Le Canal du Midi on the list of world’s legacy. The first plane trees bordering the channel appeared lately after its creation in 1776. Resolving the erosion of riverbanks and the evaporation of the water, the plane imposes itself as a dominant and emblematic marrow. The wooded legacy is estimated to about 42.000 trees, it contributes to the landscape and aesthetic.<br />
Today, the canal du midi is primarily used for tourism, recreation and housing. Busier than the Seine (Paris river), it accounts for one fifth of French river tourism and 80 % tourists are foreigners. Boaters seeking for tranquility, quietness and a unique environment mainly navigate over it.<br />
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<p class="title1">Canker Stain</p> <br />
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<p class="texte">The canker stain is a disease caused by <i>Ceratocystis platani</i>, a microscopic fungus that exclusively attacks Plane Trees.<br />
Probably introduced in France by infected wooden munitions cases coming from United States of America in 1945, the fungus introduces itself inside the sain tree’s heart, blocks the sap flow and kills it in only 2 to 5 years.<br />
On the regulatory side, the July 31th 2000 modified ministerial order classes <i>Ceratocystis</i> as a harmful organism for plants. This order leads to a mandatory fight against the fungus.<br />
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<p class="title1">Fighting the plane trees pathogen</p> <br />
<p class="texte">Regarding this imminent threat, some solutions emerged.<br />
</br>First of all, chemical fungicides were used. However, the community realized it was impossible to eradicate the whole fungus. <br />
</br>Therefore, the French Institut National de Recherche Agronomique (INRA) created a new type of plane trees named Platanor which were resistant to the fungus infection. Unfortunately, this extreme measure could not save the already contaminated trees nearby the Canal du Midi.<br />
</br>Today, the only option remains in the tree-cutting of the contaminated plane trees. Nevertheless, this previous solution is not optimal because of the high cost and the major ecological, touristic and social impact for our heritage.<br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/TeamTeam:Toulouse/Team2014-10-16T20:08:44Z<p>Laureen: </p>
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<p style="color:#696969; padding-top:20px; font-size:16px; float:left;"> Team&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Our Team</p> <br />
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<p class="texte">Let us introduce you to our fabulous tree-friendly team , 100% biodegradable and certified pesticide-free.</p><br />
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<p class="title1">Students</p><br />
<p class="title3"> <i>(Put the mouse over the pictures to extend them!)</i> </p><br />
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<p class="title2"><b>Diane Barbay</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Diligent (Wait a minute, I have to write down these results in my notebook! ㅋㅋㅋ)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About her:</b> Hardworking, Diane is the most conscientious in her work: her notebook is so clean and organized! She is persevering and always tries to find the answer to the problem (except when plasmids shortens between two digestions…). She talks to bacteria to encourage them taking up plasmid during transformation. She is like a babysitter for our cells :-)<br />
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<p class="title2"><b>Emeline Flajollet</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Expert in competent cells (Oh no, I have to make competent cells again…)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Master of Microbiology at Paul Sabatier University of Toulouse</p> <br />
<p class="textesimple"><b>About her:</b> Emeline looks discreet at first, but when you meet her, you discover a frank person who knows how to express her opinions and how to be understood, which is a good thing for team work. She is diligent and involved in her work, always trying to do her best, and providing advice to others. She spends a lot of time on social networks, allowing us to keep in touch with other teams.<br />
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<p class="title2"><b>Mathieu Fournié</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Expert in communication (I found someone else who wants to interview us!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Master of Microbiology at Paul Sabatier University of Toulouse</p> <br />
<p class="textesimple"><b>About him:</b> Initial founder of the project, Mathieu is a fan of Midi Pyrenées's region and is eager to protect its natural environment and resources. He is devoted to the project and does his best to succeed. His leadership skills help us focus on the main goals and deadlines. Even if it makes him forget to do his own tasks! His good interpersonal skills and numerous contacts helped us to capture media attention. <br />
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<p class="title2"><b>Florie Gosseau</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our IT expert (Modelling? Ok let’s do it!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Bioinformatics and Biological Systems Master at Paul Sabatier University of Toulouse</p> <br />
<p class="textesimple"><b>About her:</b> Florie is a smiling and spontaneous girl. She is this kind of person capable of giving a true smile when nothing works, and she can disentangle conflicts thanks to her patience and her kindness. She is really helpful and never says no when you ask for service. She is also frank and reports problems when there are some. Finally, she is the only one good at informatics, which makes her indispensable for us!<br />
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<p class="title2"><b>Camille Jourdan</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Expert in cloning (Camille, which enzyme should I take?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About him:</b> Jovial and funny, Camille has sometimes eruptions of madness which makes him so special. Conversely, he is capable of incredible moments of intelligence and concentration (primers and plasmids hold no secret for him) and he is really diligent. He is sarcastic but definitely not nasty. He is the athlete of the team and never misses an opportunity to make gymnastics exercises or Plasmid Dance!<br />
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<p class="title2"><b>Aurélie Kanitzer</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Distracted (Oh I forgot my keys! I have to go back home, see you in 15 minutes!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About her:</b> Aurélie is a joyful and spontaneous person, which makes you laugh and smile. Often distracted, she can make blunders… She is natural and honest and does not try to play a role. She is open-minded and helpful: you can count on her. Strong and sensitive at the same time, she is able to face difficulties. Don’t go by appearances, this tiny girl has personality and she does not let others walk over her!<br />
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<p class="title2"><b>Laureen Mirassou</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our best negotiator (Don’t worry about prices for flight tickets, I deal with that)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About her:</b> Laureen is a mix of kindness, helpfulness and generosity. Always trying to help people, she is essential to ease tensions and to solve social issues: she says what is wrong gently and calmly. Her good interpersonal abilities helped us to make good business and to solve many administrative problems. Her high level of English proficiency is also something precious for the team.<br />
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<p class="title2"><b>Manon Molina</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Organized (Let’s make a to-do-list!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school </p> <br />
<p class="textesimple"><b>About her:</b> Manon is organized: she always makes to-do-lists to not forget anything and do tasks in time. Thank goodness, she is here to monitor the budget and do the accounting! She makes her work conscientiously and meticulously, not allowing mistakes. She is kind and lenient but can also be frank when she does not endorse your methods. In short: serious and devoted to the project.<br />
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<p class="title2"><b>Fanny Pineau</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Absent-minded (which cloning are we doing?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school </p> <br />
<p class="textesimple"><b>About her:</b> In the lab, Fanny is often lost in her works: too many cloning at the same time, you should not ask too much to her blond brain! Not enough concentrated, she can sometimes make blunders. Besides that, she is applied and perfectionist when she plunges into her work. Finally, she is a joyful and smiling person; also honest and frank, telling it like it is.<br />
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<p class="title2"><b>Pierre Reitzer</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Relaxed (Make a presentation barefoot, why not?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school </p> <br />
<p class="textesimple"><b>About him:</b> Pierre is the co-founder of the project. He is really friendly and is always willing to discuss and help people with their experiments. Moreover, he greets us with a smile each time we see him in the lab. He is spontaneous and forthright: we know what he thinks. Sometimes, he has his head in the clouds and fails the same experiment three times… But he is involved in his work and does not count hours. <br />
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<p class="title2"><b>Abdel Touré</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Luxury tastes (Why would we go elsewhere than the Hilton?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Master at ENSAT Toulouse agronomic school</p> <br />
<p class="textesimple"><b>About him:</b> Abdel is this kind of person that you remember: he is always jovial and running everywhere. He is a funny guy who likes making jokes. But, be careful! He is also sensitive, your jokes might not make him laugh at all. He takes care of his appearance, and likes smoking his e-cigarette. He always swears in English, but we like it because this is funny, and he is a good English speaker!<br />
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<p class="title2"><b>Gilles Truan</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our amazing instructor!</p><br />
<p class="textesimple"><b>Job:</b> Researcher at CNRS (Centre National de la Recherche Scientifique) and at LISBP (Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés).</p> <br />
<p class="textesimple"><b>About him:</b> He is with us from the beginning to the end as a father (and already a grandfather!!), our instructor is the master of cloning and the captain of PCR! </p><br />
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<p class="title2"><b>Brice Enjalbert</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> The right-hand man of our amazing instructor!</p><br />
<p class="textesimple"><b>Job:</b> Teacher-Researcher at LISBP (Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés).</p> <br />
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<p class="title2"><b>Florence Bordes</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> The only female advisor! </p><br />
<p class="textesimple"><b>Job:</b> Teacher-Researcher at LISBP (Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés).</p> <br />
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<img src="https://static.igem.org/mediawiki/2014/2/2d/Advisor_Cam.jpg" style="margin-top:5px" /><br />
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<p class="title2"><b>Kaymeuang Cam</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our bacteriologist advisor! </p><br />
<p class="textesimple"><b>Job:</b> Teacher-Researcher at IPBS (Institut de Pharmacologie et de Biologie Structurale)</p> <br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/TeamTeam:Toulouse/Team2014-10-16T20:01:21Z<p>Laureen: </p>
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<p style="color:#696969; padding-top:20px; font-size:16px; float:left;"> Team&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Our Team</p> <br />
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<p class="texte">Let us introduce you to our fabulous tree-friendly team , 100% biodegradable and certified pesticide-free.</p><br />
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<p class="title1">Students</p><br />
<p class="title3"> <i>(Put the mouse over the pictures to extend them!)</i> </p><br />
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<p class="title2"><b>Diane Barbay</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Diligent (Wait a minute, I have to write down these results in my notebook! ㅋㅋㅋ)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About her:</b> Hardworking, Diane is the most conscientious in her work: her notebook is so clean and organized! She is persevering and always tries to find the answer to the problem (except when plasmids shortens between two digestions…). She talks to bacteria to encourage them taking up plasmid during transformation. She is like a babysitter for our cells :-)<br />
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<p class="title2"><b>Emeline Flajollet</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Expert in competent cells (Oh no, I have to make competent cells again…)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Master of Microbiology at Paul Sabatier University of Toulouse</p> <br />
<p class="textesimple"><b>About her:</b> Emeline looks discreet at first, but when you meet her, you discover a frank person who knows how to express her opinions and how to be understood, which is a good thing for team work. She is diligent and involved in her work, always trying to do her best, and providing advice to others. She spends a lot of time on social networks, allowing us to keep in touch with other teams.<br />
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<p class="title2"><b>Mathieu Fournié</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Expert in communication (I found someone else who wants to interview us!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Master of Microbiology at Paul Sabatier University of Toulouse</p> <br />
<p class="textesimple"><b>About him:</b> Initial founder of the project, Mathieu is a fan of Midi Pyrenées's region and is eager to protect its natural environment and resources. He is devoted to the project and does his best to succeed. His leadership skills help us focus on the main goals and deadlines. Even if it makes him forget to do his own tasks! His good interpersonal skills and numerous contacts helped us to capture media attention. <br />
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<p class="title2"><b>Florie Gosseau</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our IT expert (Modelling? Ok let’s do it!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Bioinformatics and Biological Systems Master at Paul Sabatier University of Toulouse</p> <br />
<p class="textesimple"><b>About her:</b> Florie is a smiling and spontaneous girl. She is this kind of person capable of giving a true smile when nothing works, and she can disentangle conflicts thanks to her patience and her kindness. She is really helpful and never says no when you ask for service. She is also frank and reports problems when there are some. Finally, she is the only one good at informatics, which makes her indispensable for us!<br />
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<p class="title2"><b>Camille Jourdan</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Expert in cloning (Camille, which enzyme should I take?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About him:</b> Jovial and funny, Camille has sometimes eruptions of madness which makes him so special. Conversely, he is capable of incredible moments of intelligence and concentration (primers and plasmids hold no secret for him) and he is really diligent. He is sarcastic but definitely not nasty. He is the athlete of the team and never misses an opportunity to make gymnastics exercises or Plasmid Dance!<br />
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<p class="title2"><b>Aurélie Kanitzer</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Distracted (Oh I forgot my keys! I have to go back home, see you in 15 minutes!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About her:</b> Aurélie is a joyful and spontaneous person, which makes you laugh and smile. Often distracted, she can make blunders… She is natural and honest and does not try to play a role. She is open-minded and helpful: you can count on her. Strong and sensitive at the same time, she is able to face difficulties. Don’t go by appearances, this tiny girl has personality and she does not let others walk over her!<br />
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<p class="title2"><b>Laureen Mirassou</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our best negotiator (Don’t worry about prices for flight tickets, I deal with that)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About her:</b> Laureen is a mix of kindness, helpfulness and generosity. Always trying to help people, she is essential to ease tensions and to solve social issues: she says what is wrong gently and calmly. Her good interpersonal abilities helped us to make good business and to solve many administrative problems. Her high level of English proficiency is also something precious for the team.<br />
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<p class="title2"><b>Manon Molina</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Organized (Let’s make a to-do-list!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school </p> <br />
<p class="textesimple"><b>About her:</b> Manon is organized: she always makes to-do-lists to not forget anything and do tasks in time. Thank goodness, she is here to monitor the budget and do the accounting! She makes her work conscientiously and meticulously, not allowing mistakes. She is kind and lenient but can also be frank when she does not endorse your methods. In short: serious and devoted to the project.<br />
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<p class="title2"><b>Fanny Pineau</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Absent-minded (which cloning are we doing?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school </p> <br />
<p class="textesimple"><b>About her:</b> In the lab, Fanny is often lost in her works: too many cloning at the same time, you should not ask too much to her blond brain! Not enough concentrated, she can sometimes make blunders. Besides that, she is applied and perfectionist when she plunges into her work. Finally, she is a joyful and smiling person; also honest and frank, telling it like it is.<br />
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<p class="title2"><b>Pierre Reitzer</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Relaxed (Make a presentation barefoot, why not?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school </p> <br />
<p class="textesimple"><b>About him:</b> Pierre is the co-founder of the project. He is really friendly and is always willing to discuss and help people with their experiments. Moreover, he greets us with a smile each time we see him in the lab. He is spontaneous and forthright: we know what he thinks. Sometimes, he has his head in the clouds and fails the same experiment three times… But he is involved in his work and does not count hours. <br />
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<p class="title2"><b>Abdel Touré</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Luxury tastes (Why would we go elsewhere than the Hilton?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Master at ENSAT Toulouse agronomic school</p> <br />
<p class="textesimple"><b>About him:</b> Abdel is this kind of person that you remember: he is always jovial and running everywhere. He is a funny guy who likes making jokes. But, be careful! He is also sensitive, your jokes might not make him laugh at all. He takes care of his appearance, and likes smoking his e-cigarette. He always swears in English, but we like it because this is funny, and he is a good English speaker!<br />
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<p class="title1">Instructors</p><br />
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<p class="title2"><b>Gilles Truan</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our amazing instructor!</p><br />
<p class="textesimple"><b>Job:</b> Researcher at CNRS (Centre National de la Recherche Scientifique) and at LISBP (Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés).</p> <br />
<p class="textesimple"><b>About him:</b> With us from the beginning to the end as a father (and already a grandfather!!), our instructor is the master of cloning and the captain of PCR! </p><br />
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<p class="title2"><b>Brice Enjalbert</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> The right-hand man of our amazing instructor!</p><br />
<p class="textesimple"><b>Job:</b> Teacher-Researcher at LISBP (Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés).</p> <br />
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<p class="title2"><b>Florence Bordes</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> The only female advisor! </p><br />
<p class="textesimple"><b>Job:</b> Teacher-Researcher at LISBP (Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés).</p> <br />
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<p class="title2"><b>Kaymeuang Cam</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our bacteriologist advisor! </p><br />
<p class="textesimple"><b>Job:</b> Teacher-Researcher at IPBS (Institut de Pharmacologie et de Biologie Structurale)</p> <br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/TeamTeam:Toulouse/Team2014-10-16T20:00:25Z<p>Laureen: </p>
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<p style="color:#696969; padding-top:20px; font-size:16px; float:left;"> Team&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Our Team</p> <br />
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<p class="texte">Let us introduce you to our fabulous tree-friendly team , 100% biodegradable and certified pesticide-free.</p><br />
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<p class="title1">Students</p><br />
<p class="title3"> <i>(Put the mouse over the pictures to extend them!)</i> </p><br />
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<p class="title2"><b>Diane Barbay</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Diligent (Wait a minute, I have to write down these results in my notebook! ㅋㅋㅋ)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About her:</b> Hardworking, Diane is the most conscientious in her work: her notebook is so clean and organized! She is persevering and always tries to find the answer to the problem (except when plasmids shortens between two digestions…). She talks to bacteria to encourage them taking up plasmid during transformation. She is like a babysitter for our cells :-)<br />
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<p class="title2"><b>Emeline Flajollet</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Expert in competent cells (Oh no, i have to make competent cells again… ㅋㅋㅋ)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Master of Microbiology at Paul Sabatier University of Toulouse</p> <br />
<p class="textesimple"><b>About her:</b> Emeline looks discreet at first, but when you meet her, you discover a frank person who knows how to express her opinions and how to be understood, which is a good thing for team work. She is diligent and involved in her work, always trying to do her best, and providing advice to others. She spends a lot of time on social networks, allowing us to keep in touch with other teams.<br />
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<p class="title2"><b>Mathieu Fournié</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Expert in communication (I found someone else who wants to interview us!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Master of Microbiology at Paul Sabatier University of Toulouse</p> <br />
<p class="textesimple"><b>About him:</b> Initial founder of the project, Mathieu is a fan of Midi Pyrenées's region and is eager to protect its natural environment and resources. He is devoted to the project and does his best to succeed. His leadership skills help us focus on the main goals and deadlines. Even if it makes him forget to do his own tasks! His good interpersonal skills and numerous contacts helped us to capture media attention. <br />
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<p class="title2"><b>Florie Gosseau</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our IT expert (Modelling? Ok let’s do it!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Bioinformatics and Biological Systems Master at Paul Sabatier University of Toulouse</p> <br />
<p class="textesimple"><b>About her:</b> Florie is a smiling and spontaneous girl. She is this kind of person capable of giving a true smile when nothing works, and she can disentangle conflicts thanks to her patience and her kindness. She is really helpful and never says no when you ask for service. She is also frank and reports problems when there are some. Finally, she is the only one good at informatics, which makes her indispensable for us!<br />
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<p class="title2"><b>Camille Jourdan</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Expert in cloning (Camille, which enzyme should I take?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About him:</b> Jovial and funny, Camille has sometimes eruptions of madness which makes him so special. Conversely, he is capable of incredible moments of intelligence and concentration (primers and plasmids hold no secret for him) and he is really diligent. He is sarcastic but definitely not nasty. He is the athlete of the team and never misses an opportunity to make gymnastics exercises or Plasmid Dance!<br />
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<p class="title2"><b>Aurélie Kanitzer</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Distracted (Oh I forgot my keys! I have to go back home, see you in 15 minutes!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About her:</b> Aurélie is a joyful and spontaneous person, which makes you laugh and smile. Often distracted, she can make blunders… She is natural and honest and does not try to play a role. She is open-minded and helpful: you can count on her. Strong and sensitive at the same time, she is able to face difficulties. Don’t go by appearances, this tiny girl has personality and she does not let others walk over her!<br />
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<p class="title2"><b>Laureen Mirassou</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our best negotiator (Don’t worry about prices for flight tickets, I deal with that)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school</p> <br />
<p class="textesimple"><b>About her:</b> Laureen is a mix of kindness, helpfulness and generosity. Always trying to help people, she is essential to ease tensions and to solve social issues: she says what is wrong gently and calmly. Her good interpersonal abilities helped us to make good business and to solve many administrative problems. Her high level of English proficiency is also something precious for the team.<br />
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<p class="title2"><b>Manon Molina</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Organized (Let’s make a to-do-list!)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school </p> <br />
<p class="textesimple"><b>About her:</b> Manon is organized: she always makes to-do-lists to not forget anything and do tasks in time. Thank goodness, she is here to monitor the budget and do the accounting! She makes her work conscientiously and meticulously, not allowing mistakes. She is kind and lenient but can also be frank when she does not endorse your methods. In short: serious and devoted to the project.<br />
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<p class="title2"><b>Fanny Pineau</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Absent-minded (which cloning are we doing?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school </p> <br />
<p class="textesimple"><b>About her:</b> In the lab, Fanny is often lost in her works: too many cloning at the same time, you should not ask too much to her blond brain! Not enough concentrated, she can sometimes make blunders. Besides that, she is applied and perfectionist when she plunges into her work. Finally, she is a joyful and smiling person; also honest and frank, telling it like it is.<br />
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<p class="title2"><b>Pierre Reitzer</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Relaxed (Make a presentation barefoot, why not?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Biochemistry Master at INSA Toulouse engineering school </p> <br />
<p class="textesimple"><b>About him:</b> Pierre is the co-founder of the project. He is really friendly and is always willing to discuss and help people with their experiments. Moreover, he greets us with a smile each time we see him in the lab. He is spontaneous and forthright: we know what he thinks. Sometimes, he has his head in the clouds and fails the same experiment three times… But he is involved in his work and does not count hours. <br />
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<p class="title2"><b>Abdel Touré</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Luxury tastes (Why would we go elsewhere than the Hilton?)</p><br />
<p class="textesimple"><b>Studies:</b> First year of Master at ENSAT Toulouse agronomic school</p> <br />
<p class="textesimple"><b>About him:</b> Abdel is this kind of person that you remember: he is always jovial and running everywhere. He is a funny guy who likes making jokes. But, be careful! He is also sensitive, your jokes might not make him laugh at all. He takes care of his appearance, and likes smoking his e-cigarette. He always swears in English, but we like it because this is funny, and he is a good English speaker!<br />
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<p class="title2"><b>Gilles Truan</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our amazing instructor!</p><br />
<p class="textesimple"><b>Job:</b> Researcher at CNRS (Centre National de la Recherche Scientifique) and at LISBP (Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés).</p> <br />
<p class="textesimple"><b>About him:</b> With us from the beginning to the end as a father (and already a grandfather!!), our instructor is the master of cloning and the captain of PCR! </p><br />
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<p class="title2"><b>Brice Enjalbert</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> The right-hand man of our amazing instructor!</p><br />
<p class="textesimple"><b>Job:</b> Teacher-Researcher at LISBP (Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés).</p> <br />
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<p class="title2"><b>Florence Bordes</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> The only female advisor! </p><br />
<p class="textesimple"><b>Job:</b> Teacher-Researcher at LISBP (Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés).</p> <br />
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<p class="title2"><b>Kaymeuang Cam</CENTER></b></p><br />
<p class="textesimple"><b>Feature:</b> Our bacteriologist advisor! </p><br />
<p class="textesimple"><b>Job:</b> Teacher-Researcher at IPBS (Institut de Pharmacologie et de Biologie Structurale)</p> <br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:ToulouseTeam:Toulouse2014-10-16T19:51:31Z<p>Laureen: </p>
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<!-------------------------------- CONTENT ---------------------------------><br />
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<img src="https://static.igem.org/mediawiki/2014/b/b6/Canal_du_Midi_-_plane_trees.jpeg" alt="Présentation igem toulouse" /><br />
<div class="text-presentation"><br />
<h1 class="title1" style="font-size:28px;"><br />
SubtiTree project<br />
</h1><br />
<p class="texte"><br />
Fungal diseases lead to major economic losses all over the world. Some pathogens trigger tracheomycosis by disturbing the vascular system of the plant. Canker is one of these infections especially affecting plane trees (<i>Platanus sp.</i>). Plane trees are widely present in Southern France and in particular along the famous Canal du Midi, participating to the site gorgeousness. Today, the only treatment consists in a costly preventive tree-cutting and implies significant ecological troubles. Facing this emergency, our team offers an innovative solution originating from synthetic biology. <br />
</p><br />
<a class="button-home" href="https://2014.igem.org/Team:Toulouse/Project/project-context" style="border: 1px solid #282828;-webkit-border-radius: 5px;-moz-border-radius: 5px;border-radius: 5px; padding: 13px 25px 12px; color: #282828; text-decoration: none; font-size: 17px; background: none; display: block; width: 89px;">Learn more</a><br />
</div><br />
</div><br />
<br />
<div class="clear"></div><br />
<br />
<div class="gallerie-project" style="margin-bottom:50px; margin-top:70px;"><br />
<br />
<div class="line-gallerie-project"><br />
<a href="https://2014.igem.org/Team:Toulouse/Project/Overviews" class="part-little-projet"><br />
<div class="title-part-projet">Overview</div><br />
<div style="position:absolute; top:90px; left:34px; color:white; font-family:'Open Sans'; font-size:16px; width:200px; line-height:24px;">Let's save our trees with SubtiTree!</div><br />
<div style="border-bottom: 1px solid #fff; color:white; font-family:'Open Sans'; position:absolute; bottom:25px; right:22px; font-size:16px;">Read more</div><br />
</a><br />
<br />
<a href="https://2014.igem.org/Team:Toulouse/Project/Chemotaxis" class="part-large-projet" style="background: url('https://static.igem.org/mediawiki/2014/a/aa/Chimio.jpg') no-repeat center;-webkit-background-size: cover;-moz-background-size: cover;-o-background-size: cover;background-size: cover;"><br />
<div class="title-part-projet" style="color:white; ">Chemotaxis</div><br />
<div style="position:absolute; top:90px; left:50px; color:white; font-family:'Open Sans'; font-size:16px; width:200px; line-height:24px;">To target the pathogenic fungus</div><br />
<div style="border-bottom: 1px solid #white; color:white; font-family:'Open Sans'; position:absolute; bottom:25px; left:50px; font-size:16px;"><u>Read more</u></div><br />
</a><br />
</div><br />
<br />
<div class="line-gallerie-project"><br />
<a href="https://2014.igem.org/Team:Toulouse/Project/binding" class="part-large-projet" style="background: url('https://static.igem.org/mediawiki/2014/0/0b/Billes3D.png') no-repeat center;-webkit-background-size: cover;-moz-background-size: cover;-o-background-size: cover;background-size: cover;"><br />
<div class="title-part-projet style="color:white; ">Binding</div><br />
<div style="position:absolute; top:90px; left:34px; color:white; font-family:'Open Sans'; font-size:16px; width:200px; line-height:24px">To be attached to the fungal pathogen wall</div><br />
<div style="border-bottom: 1px solid #515553; color:white; font-family:'Open Sans'; position:absolute; bottom:28px; right:28px; font-size:16px;">Read more</div><br />
</a><br />
<br />
<a href="https://2014.igem.org/Team:Toulouse/Modelling" class="part-little-projet"><br />
<div class="title-part-projet">Modeling</div><br />
<div style="position:absolute; top:90px; left:34px; color:white; font-family:'Open Sans'; font-size:16px; width:200px; line-height:24px">To develop a predictive model</div><br />
<div style="border-bottom: 1px solid #fff; color:white; font-family:'Open Sans'; position:absolute; bottom:28px; right:22px; font-size:16px;">Read more</div><br />
</a><br />
</div><br />
<br />
<div class="line-gallerie-project"><br />
<a href="https://2014.igem.org/Team:Toulouse/Project/Spreading" class="part-little-projet"><br />
<div class="title-part-projet">Spreading</div><br />
<div style="position:absolute; top:90px; left:34px; color:white; font-family:'Open Sans'; font-size:16px; width:200px; line-height:24px">How do we keep control on Subtitree?</div><br />
<div style="border-bottom: 1px solid #fff; color:white; font-family:'Open Sans'; position:absolute; bottom:25px; right:22px; font-size:16px;"">Read more</div><br />
</a><br />
<br />
<a href="https://2014.igem.org/Team:Toulouse/Project/Fungicides" class="part-large-projet" style="background:url('https://static.igem.org/mediawiki/2014/3/38/Fungi.jpg') no-repeat center;-webkit-background-size: cover;-moz-background-size: cover;-o-background-size: cover;background-size: cover;"><br />
<div class="title-part-projet" style="color:white;">Fungicides</div><br />
<div style="position:absolute; top:90px; left:50px; color:white; font-family:'Open Sans'; font-size:16px; width:200px; line-height:24px;">To eradicate fungal diseases</div><br />
<div style="border-bottom: 1px solid white; color:white; font-family:'Open Sans'; position:absolute; bottom:25px; right:22px; font-size:16px;">Read more</div><br />
</a><br />
</div><br />
<br />
</div><br />
<br />
</div><br />
<br />
</div><br />
<br />
</div><br />
<br />
<br />
<!-------------------------------- ASIDE ---------------------------------><br />
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<br />
<div class="wrapper-aside"><br />
<br />
<h2 style="color:green; text-align:center; border:none; font-weight:bold; margin:30px 0px 50px 0px;">HUMAN PRACTICE</h2><br />
<br />
<div class="sticker-aside" style="margin-right:20px;"><br />
<img src="https://static.igem.org/mediawiki/2014/6/67/Template-igem2014-HP-1.jpg" alt="image safety" /><br />
<h2>Safety</h2><br />
<p>Safety is not just a slogan, it is a way of life.</p><br />
<a href="https://2014.igem.org/Team:Toulouse/Safety"><img src="https://static.igem.org/mediawiki/2014/6/6f/Template-igem2014-iconeHP.png"/></a><br />
</div><br />
<br />
<div class="sticker-aside" style="margin-right:20px;"><br />
<img src="https://static.igem.org/mediawiki/2014/e/ea/Ethics.jpg" alt="" /><br />
<h2>Ethics</h2><br />
<p>Ethics are more important than laws.</p><br />
<a href="https://2014.igem.org/Team:Toulouse/ethics"><img src="https://static.igem.org/mediawiki/2014/6/6f/Template-igem2014-iconeHP.png"/></a><br />
</div><br />
<br />
<div class="sticker-aside" style="position: relative;bottom:19px;"><br />
<img src="https://static.igem.org/mediawiki/2014/2/20/Bacteria_talk.png" alt="" style="margin-top:26px;" /><br />
<h2>Communication</h2><br />
<p>Good words are worth much, and cost little.</p><br />
<a href="https://2014.igem.org/Team:Toulouse/Communication"><img src="https://static.igem.org/mediawiki/2014/6/6f/Template-igem2014-iconeHP.png"/></a><br />
</div><br />
<br />
</div> <br />
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<!-------------------------------- SPONSOR ---------------------------------><br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/Result/experimental-resultsTeam:Toulouse/Result/experimental-results2014-10-16T19:41:08Z<p>Laureen: </p>
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<h2>Experimental results</h2><br />
<p> Are our modules functionnal? </p><br />
</div><br />
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<br />
<div class="fils-ariane" style="width:100%; height:60px; background:#ededed;"><br />
<p style="margin:0 auto; color:#696969; width:960px; padding-top:20px; font-size:16px;"> Results&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Experimental results</p> <br />
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<br />
<p class="texte">What were the results of our experiments ? Click on these next titles to see SubtiTree abilities.</p><br />
<br />
<p style="font-size:1.3em; margin: 0 0 30px 0;"><a href="#" onClick="ddaccordion.collapseall('technology'); return false">Collapse all</a> | <a href="#" onClick="ddaccordion.expandall('technology'); return false">Expand all</a></p><br />
<br />
<div class="technology">Chemotaxis</div><br />
<div class="thelanguage"><br />
<br />
<br />
<p class="texte"> We performed several tests to demonstrate the chemotaxis ability of the transformed <i>Bacillus subtilis</i> strain towards NAG and we used the wild type bacteria and glucose as a positive glucose. <br />
</p><br />
<p class="title2">1. Petri Dishes Test </p><br />
<br />
<p class="texte"> We first tried to test chemotaxis onto Petri Dishes filled with a 0.3% agar medium. This semi-solid medium allows the bacterial motility. A paper disk containing an attractive compound is placed in the middle of the dish and cells are then loaded in the medium (see Figure 1). This protocol was taken from the <a href="https://2011.igem.org/Team:Imperial_College_London/Protocols_Chemotaxis">the Imperial College 2011 iGEM team</a>.</p><br />
<br />
<center><img SRC="https://static.igem.org/mediawiki/2014/0/05/Schema_1.png" alt="schema Figure 1" style="width:500px"></center><br />
<p class="legend">Figure 1: Schema showing how cells are filled in the medium. (A) Pipettes are used to put cells in the medium. (B) Bacteria should move to the attractive compound which diffuses.</p><br />
<br />
<p class="texte">We did not have any result with WT <i>Bacillus subtilis</i> and glucose as attractive compound (Figure 2-A). <i>B. subtilis</i> is attracted by many other glucides and amino-acids, so we also tried to test diluted glucose in LB medium attractant (Figure 2-B).</p><br />
<br />
<br />
<center><img SRC="https://static.igem.org/mediawiki/2014/f/ff/Fig2_AetB.png" alt="Figure 2" style="width:750px"></center><br />
<p class="legend">Figure 2: Chemotaxis test with Glucose as attractive compound (A) and Glucose added to LB medium as attractant (B).</p><br />
<br />
<p class="texte"> We could not notice any difference between the petri dish with or without glucose. With an addition of LB medium to sugar, a large halo around the paper disk was noticeable. This halo may correspond to cells attracted by the solution, as it is not noticeable when cells are not added (data not shown). Anyway we did not have enough reproducible and reliable results to be satisfied with this test.<br> <br />
Furthermore, with the addition of LB medium, it is hard to make the distinction between the attractive effects and the simple growth resulting from random diffusion.</br><br />
We have started new tries using different protocols.</p><br />
<br />
<p class="title2">2. Plug in Pond system<br />
</p><br />
<br />
<p class="texte"><br />
This protocol we worked on is taken from a thesis (see references [1]). . A solution of <i>B.subtilis</i> is grown overnight so as to obtain a cell density of 8x10⁸ cells/mL. 10mL of the solution is mixed with 15mL of LB medium with 1.5 % agar kept at 45°.The final concentration of the obtained medium is 0.9% agar. Tetracycline is aded at 25µg/mL, in order to inhibit growth and to only observe the chemotaxis phenomenon. Plates are cooled and dried, before digging wells with a punch or 1mL tips. The wells are filled with attractive compounds (Figure 3). After one hour at room temperature, photos of the plates are taken and the results are analyzed.<br />
</p><br />
<br />
<center><img SRC="https://static.igem.org/mediawiki/2014/c/cd/Fig3.png" alt="Figure 3" style="width:400px"></center><br />
<p class="legend">Figure 3: Schema showing how are made plug-in-pond tests.</p><br />
<br />
<p class="texte"><br />
On our first try with <i>B. subtilis</i>, we made three wells per plate (Figure 4).The wells were filled with glucose at different concentrations and tetracycline was not added in one of the plates.<br />
</p><br />
<br />
<center><img SRC="https://static.igem.org/mediawiki/2014/c/ce/Fig4.png" alt="Figure 4" style="width:400px"></center><br />
<p class="legend">Figure 4: Plates after 12h at room temperature.</p><br />
<br />
<p class="texte"><br />
After an hour, no tangible results were obtained. It is only after 12hours that we were able to observe halos around the wells with glucose at 1M in the plates without tetracycline. Tetracycline concentration seems to be too high and inhibits any bacterial activity. Therfore, we have worked with tetracycline at 15µg/mL.<br />
We tried this protocol again with this new condition. We made two wells per plate (Figure 5), one with either Glucose or N-acetyl-glucosamine and one with LB medium. As previsously, no results were achieved after 1h, but after 12hours we could notice halos.<br />
</p><br />
<br />
<center><img SRC="https://static.igem.org/mediawiki/2014/c/c3/Bsubtilis_result.png" alt="Figure 5" style="width:750px"></center><br />
<p class="legend">Figure 5: Chemotaxis test with <i>Bacillus subtilis</i> WT. The upper wells contain attractive compound and the lower contain medium without attractive compound. </p><br />
<br />
<p class="texte"><br />
Results are not as clear as the first time, but we observed halos around the well with glucose at 250mM with and without tetracycline. We have then tried the same experiment with N-acetyl-glucosamine and we did not see any halo in the tested conditions. Thus we assumed that our <i>B. subtilis</i> 168 strain was not attracted to N-acetyl-glucosamine.<br />
However, the results are not clear, reliable and reproducible enough with the plug-in-pond protocol. Another testing protocol was then adopted. <br />
<br />
</p><br />
<br />
<br />
<p class="texte"><br />
<b>References:</b></br><br />
[1] : Etude de la réponse adaptative à l'oxyde de triméthylamine et de son mécanisme de détection chez Escherichia coli et Shewanella oneidensis, 2008, Claudine Baraquet, université de la méditerranée Aix-Marseille II<br />
</p><br />
<br />
<br />
<br />
<br />
<p class="title2">4. Capillary test between two tubes also called the tubes test</p><br />
<p class="texte">After the experiment of the plug-in-pond, we decided to construct a system by welding two Eppendorf tubes with a capillary thanks to an electric burner.</p><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2014/f/fb/Chemotaxis_-_eppendorf.png"></center><br />
<p class="legend">Figure 6: Photography of the first tubes system</p><br />
<br />
<p class="texte">We tested this system with a fuchsin dye and water and we were able to observe the diffusion of fuchsin towards water. However this construction had a leakage next to the weld seam that we could not stop. <br />
Thus, we asked the help from the INSA glass blower, Patrick Chekroun. He designed two systems composed of two tubes linked by a capillary.</p><br />
<br />
<br />
<center><img src="https://static.igem.org/mediawiki/2014/2/2b/Chemotaxis_-_tubes.png"><center><br />
<p class="legend">Figure 7: Scheme of the tubes system</p><br />
<br />
<p class="texte">As we did previously, we tested this new system with fuchsin. This experiment was made with WT <i>Bacillus subtilis</i> and N-Acetylglucosamine.<br />
<br><br><br />
<i>NB: We could not see the diffusion from one tube to the other. We made the hypothesis that it was not visible by sight because of the small diameter of the capillary. <br />
</i><br><br />
<br><br />
The following strategy was used to avoid disturbance due to pressure and liquid movement through the capillary:<br><br />
- The first step was the addition of Wash Buffer until the capillary was full to avoid the presence of air bubbles which could lead to diffusion problems.<br><br />
- Then, the tube 2 was plugged with the thumb while another person was adding the bacterial solution of WT <i>Bacillus subtilis</i> in the tube 1. <br><br />
- The tube 1 was also plugged and only after the thumb could be removed from the tube 2. <br><br />
- In the same way, the N-Acetylglucosamine was added in the tube 2. <br><br />
- The same process was made with a xylose positive control.<br><br />
<br><br />
<i>NB: According to the article Chemotaxis towards sugars by </i>Bacillus subtilis, (George W. Ordal et al., 1979), <i>glucose and xylose have the same attractant power. We prefer a positive control instead of a negative one because we were not sure that this system was efficient.</i><br><br />
<br><br />
- The system was kept straight for 2hours. Every 40 minutes, we took a sample of each tube and spread it on an agar plate (dilution 1/1,000).</p><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2014/1/1b/Chemotaxis_-_tubes_photo.png"></center><br />
<p class="legend">Figure 8: Photography of the tubes system</p><br />
<br />
<br />
<p class="texte">Unfortunately, the dilution was too high to detect any chemotaxis movement and the time was too short. We did not find any information in the literature.<br><br />
As we did not have the time to optimize this protocol we preferred using the protocol of the 2011 Imperial college iGEM team : the tips capillary test.</br><br />
</p><br />
<br />
<p class="title2"> 5. Tips capillary system</p><br />
<p class="title3">First tips capillary system</p><br />
<p class="texte">This protocol comes from 2011 Imperial College iGEM team and was adapted by our team in several steps (See <a href="https://2014.igem.org/Team:Toulouse/Notebook/Protocols#select8">chemotaxis protocol</a>).<br />
<br><br />
In the first tips capillary system, we used parafilm to avoid any kind of air disturbance in the tips. The different steps are described below:<br><br />
- 15µL of each chemo-attractant was pipetted. <br><br />
- The bottom of tip with the pipette was then put on a piece of parafilm and the pipette was removed from the top of the tip.<br><br />
- The top of the tip was then sealed with a piece of parafilm. By this way, the sterility can be assured and the liquid stays inside the tip. <br><br />
- To finish, the level of the solution in the tip was marked.<br></p><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2014/9/94/Chemotaxis_-_tip.png"></center><br />
<p class="legend">Figure 9: Sealing of a tip with parafilm</p><br />
<br />
<p class="texte">- After all the attractants were added in the tips, we put them on a green base to carry them. The whole process can be seen on Figure 10.<br><br />
- Each tip was immersed in 300 µL of a bacterial solution in the wells of an Elisa plate.<br></p><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2014/0/05/Chemotaxis_-_tip_and_support.png"></center><br />
<p class="legend">Figure 10: First tips capillary system</p><br />
<br />
<p class="texte"><i>NB: the yellow carton was used to stabilize the system and keep it straight.</i><br><br />
<br><br />
- After one hour, the tips were removed from the bacteria solutions and the content of the tips was observed with a Thoma cell under the microscope.<br><br />
<br><br />
We had several problems with this system:<br><br />
- The liquid level decreased during the experiment and we did not have enough liquid to fill the Thoma cell. Thus, it was not possible to count.<br><br />
- The bacteria were moving and therefore, we could not proceed to a bacteria count.<br><br />
<br><br />
Regarding these observations we decided to spread the tips content on agar plates instead of using Thoma cell and microscopy.<br><br />
<p class="title3">Second tips capillary system<br />
</p><br />
<p class="texte"And then the revolution came! We found a multichannel pipette :D The same protocol was performed except that the parafilm was used to avoid the air entrance between the tips and the pipette and therefore the loss of liquid.<br></p><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2014/e/e4/Chemotaxis_-_pipette.png"></center><br />
<p class="legend">Figure 11: Second tips capillary system</p><br />
<br />
<p class="title3">Improvement of the second tips capillary system</p><br />
<p class="texte">However this system was not optimal it is why we decided to use blu tack instead of parafilm: <br></p><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2014/4/42/Chemotaxis_-_pipette_and_blu_tack.png"></center><br />
<p class="legend">Figure 12: Improvement of the second tips capillary system</p><br />
<br />
<p class="texte"><b>At that point, the protocol was approved and the final test could finally start! :-)</b><br><br />
<br><br />
There was just one tiny problem… we did not have our optimized bacterium transformed with the chemotaxis module!!! That is why we concentrated our efforts on WT <i>Bacillus subtilis</i> strain.<br><br />
<br><br />
The main goal was to find an optimized control and to analyze the eventual chemotaxis of the WT strain. To avoid osmolality bias, we wanted to find a molecule which was non-attractant and with a similar molecular weight than the N-Acetylglucosamine (221.21 g/mol). Our first idea was to use fuchsin (Molecular weight: 337.85 g/mol).<br><br />
<br><br />
At the beginning, the experiment was conducted with only one negative contraol, the fuchsin and different NAG concentrations: 25mM, 250mM and 500mM. The tested strain was <i>Bacillus subtilis </i>168:<br><br />
<br></p><br />
<center><br />
<table align="center"><br />
<tr><td align=center><img src="https://static.igem.org/mediawiki/2014/8/8c/Chemotaxis_-_results_fuch.png"></td><br />
<td align=center><img src="https://static.igem.org/mediawiki/2014/f/fd/Chemotaxis_-_results_fuchsin.png"></td></tr><br />
<tr><td align=center><p class="legend">Figure 13: Fuchsin - negative control (dilution 1/50)</p></td><br />
<td align=center><p class="legend">Figure 14: NAG (25mM) (dilution 1/50)</p></td></tr><br />
</table></center><br><br />
<p class="texte">The average number of colonies with the negative control is 121. On the contrary, a cell layer is observed for the NAG plates with every concentration.<br><br />
<br><br />
Thus, we assumed that WT <i>Bacillus subtilis</i> was more attracted by NAG than fuchsin. Indeed we can neglect the bacterial growth because the test only lasts one hour. We also neglect diffusion and osmolality phenomena for the previous reasons. <br><br />
<br><br />
Unfortunately for us we forgot one major effect… Can you believe that fuchsin solution contains about 15% of ethanol?!!! This concentration can lead to the death of some cells which probably happened to our results.<br><br />
<br><br />
<b><p class="texte">This incredible and dramatic discovery destroyed all of our hopes about the God of chemotaxis! :-(</b><br><br />
<br><br />
However, our team did not give up on synthetic biology ! :-) Indeed, after days of disappointment and no time left for lab work, we raised from ashes and tried to find another negative control.<br><br />
<br><br />
Hopefully, we managed to find a negative control: galactose (25mM). The article Chemotaxis towards sugars by <i>Bacillus subtilis</i> (<i>George W. Ordal et al., 1979</i>) proved that it was a poor attractant.<br><br />
<br><br />
We made our tests again with this new molecule and glucose (25mM) as positive control.<br></p><br />
<br />
<center><img src="https://static.igem.org/mediawiki/2014/8/86/Chemotaxis_-_final_results.png"></center><br />
<p class="legend">Figure 15: Final results (dilution : 1/10,000)</p><br />
<p class="texte"> The miracle arrived! We managed to prove that our WT <i>Bacillus subtilis</i> was indeed naturally attracted to NAG.</p><br />
<p class="texte"><i>NB: It was our last experiment. Unfortunately we were running out of time and we could not do much more test. We would like to do the experiment with a lower dilution and repeat it several times.</i><br><br />
<br><br />
<b><p class="texte">Our results are not statistically significant however this result has been proved in literature.</p></b><br></p><br />
<br />
</br><br />
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<br />
<div class="technology">Binding module</div><br />
<div class="thelanguage"><br />
<br />
<br />
<p class="title2">1. Preliminary experiments</p><br />
<p class="title3">Purpose</p><br />
<p class="texte">The first experiment deals with the culture conditions to see if <i>Bacillus subtilis</i> can resist to a low temperature and with the CBB buffer. To do that, several bacterial concentrations have been tested starting with an OD of 0.1 and diluting this solution to get estimated ODs of 0.05, 0.025, 0.01. These different <i>Bacillus subtilis</i> solutions were incubated 1 hour at 4°C with 500µL of CBB or water. Finally a cell count on Thoma cell counting chamber was performed.</p><br />
<br />
<p class="title3">Results</p><br />
<p class="texte">The bacterial solutions could not be counted because of two major problems: the too high number of bacteria with the 0.1 OD or the too low number of bacteria with the 0.01 OD. Thus, the study is mostly focused on the intermediate values (Figure 16).<br />
<br/>First of all, a same cell concentration can be noticed with the presence of CBB or water with estimated ODs of 0.05 or 0.025. Moreover, twice less cells can be found in the lowest concentrations in bacteria comparing to the 0.05 OD concentration which agrees with the dilution ratio. <br />
<br/>Thus, the experimental conditions regarding the presence of CBB and the incubation temperature at 4°C do not harm the cell surviving.<br />
</p><br />
<br />
</br><br />
<center><img src="https://static.igem.org/mediawiki/2014/c/ce/Graphe_binding_1.png" width="45%"></center><br />
<br />
</br><br />
<p class="legend">Figure 16: CBB presence has no effect on bacteria. The bacterial concentration was measured regarding <span style="color:#0000FF">the presence</span> or <span style="color:#FF0000">the absence </span>of CBB for the observed OD (0.1) or estimated ODs (0.05, 0.025, 0.01).<br />
</p><br />
<br />
<p class="title2">2. Binding test using engineered <i>B. subtilis</i></p><br />
<br />
<p class="title3">Purpose</p><br />
<p class="texte">Transformed <i>Bacillus subtilis</i> with the binding module is able to produce a protein composed of the bacterial peptidoglycan bonding of LycT and the GbpA 4th domain of <i>Vibrio cholerae</i> allowing the chitin bonding. The synthetic bacterium is put with special beads composed of the polymer miming the fungal pathogen wall. After several washes, bacteria specifically attached to the chitin are put on plates and counted.</p><br />
<br />
<p class="title3">Results</p><br />
<p class="texte">The first observation is that both bacterial solutions of wild type <i>Bacillus subtilis</i> and SubtiTree have the same concentration : 105 bacteria/mL (Figure 17). Even though there is no significant difference between both strains after the first wash, the second wash has a major effect since it allows 40 times more Wild Type bacteria to come off the beads. This result correlates with the number of bacteria binded to the beads for the synthetic strain with the binding module. <br />
<br/>Thus, the binding system seems to function correctly and leads to the bacterial attachment on the chitin.</p><br />
<br />
</br><br />
<center><img src="https://static.igem.org/mediawiki/2014/e/ea/Graphe_binding_2.png" width="60%"></center><br />
</br><br />
<br />
<p class="legend">Figure 17: Attachment of <i>Bacillus subtilis</i> with binding module to chitin. <span style="color:#0000FF">The WT bacteria</span> or <span style="color:#FF0000">the bacteria with the binding system</span> concentration has been determined during the different steps of the binding test. The stars represent a significant difference observed with a Student test with p<0.05.</p><br />
<br />
<p class="title2">3. Microscopic observations</p><br />
<br />
<p class="title3">Purpose</p><br />
<p class="texte">We want to observe the SubtiTree's binding on beads coated with chitin. In order to perform a 3D reconstruction showing this interaction, we use confocal laser scanning microscope. Through the use of a fluorochrome (Syto9), we can highlight the presence of bacteria on the surface of the beads (individualized by phase-contrast). A first calibration step determined the minimum threshold to remove the background noise and the natural fluorescence.</p><br />
<br />
<p class="title3">Results</p><br />
<p class="texte">First, we note the great bacterial presence on the surface of beads coated with chitin. These images seem to highlight their interactions.</br></p><br />
<center><img src="https://static.igem.org/mediawiki/2014/archive/5/53/20141013073044!Photo_billes_microscopie.png" width="45%"></center><br />
</br><br />
<p class="legend">Figure 18: Microscopic view of beads surfaces coated with chitin</p><br />
<br />
<p class="texte">Using ImageJ software, we are able to create 3D pictures and movies of those comments.</br></p><br />
<center><img src="https://static.igem.org/mediawiki/2014/5/53/Photo_billes_microscopie.png" width="45%" style="float:left;"><iframe width="380" height="315" src="//www.youtube.com/embed/ztIHIKQr3g0" frameborder="0" allowfullscreen></iframe></center><br />
</br><br />
<p class="legend">Figure 19: A short movie of 3D bead surfaces coated with chitin</p><br />
<br />
<p class="texte">Finally we want to observe the bacteria after the second wash. When our bacterium has the binding module, results suggest a lower number of bacteria in the washing solution. SubtiTree is retained by the beads.</p><br />
<center><img src="https://static.igem.org/mediawiki/2014/9/97/Photo_lavage_microscopie.png" width="45%"></center><br />
<p class="legend">Figure 20: Microscopic view of bacteria after washing <br />
</p><br />
<br />
<p class="texte">Finally, overall results are consistent with the presence of functional binding system.</p><br />
<br />
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<br />
<div class="technology">Fungicides module</div><br />
<div class="thelanguage"><br />
<br />
<br />
<p class="title2"> 1. Preliminary experiments</p><br />
<p class="title3">Tests with commercial peptides and controls</p><br />
<p class="texte">The first tests were accomplished with commercial GAFP-1 and D4E1 peptides at different concentrations (12.5µM, 25µM, 100µM). These tests were performed on different fungal strains sharing the same phylum with <i>Ceratocystis Platani</i>.<br />
As <i>Ceratocystis Platani</i> is pathogenic, we could not perform tests directly with this fungus.</br><br />
After several days at 30°C, the PDA (Potato Dextrose Agar) plates covered with fungus and commercial peptides were analyzed.</p></p><br />
<p class="texte">An inhibition halo was noticeable with commercial D4E1 peptide at 100µM on <i>Aspergillus brasiliensis</i>. Less bright halos were also present with lower concentrations. Concerning commercial GAFP-1, we did not notice any effect in the tested conditions. As positive control, a well-known chemical fungicide was used: the Copper Sulfate. The inhibition of the fungal growth was complete at 20mg/ml, and at 10mg/ml a darker halo appeared around the pad filled with Copper Sulfate as we can see on the figure below. This corresponds to a sporulating halo in response to the stress generated by the fungicide.<br />
</p><br />
<br />
<br />
<img style="width:450px; " src="https://static.igem.org/mediawiki/parts/a/a8/Prelim_tests_fung.jpg"><br />
<img style="width:450px; " src="https://static.igem.org/mediawiki/parts/f/f8/Controls_fung.jpg"><br />
<p class="legend"> Figure 21: Results of the preliminary tests</p><br />
<br />
<br />
</br><br />
<br />
<p class="texte">Regarding these results, we concluded that very high fungicide concentrations are required to inhibit the fungal growth. Following these tests, new conditions were adopted in order not to encourage too much fungal growth over bacterial growth. The culture medium was adjusted to fit our objective and to approximate the conditions found in the trees: a 'sap-like' medium was elaborated. The incubations were then carried at room temperature.<br />
</p><br />
<br />
<p class="title2">2. Test with SubtiTree</p><br />
<br />
<p class="texte">In order to test <i>Bacillus subtilis</i> mutants, it was essential to find the right balance between the fungal growth and the bacterial one. This condition was necessary to get a high concentration of peptides. In our genetic constructions, these peptides are designed to be exported in the extracellular medium.</br><br />
</br><br />
The transformed <i>Bacillus subtilis</i> strains grew at 37°C during 72h and were tested. After centrifugation, the supernatant and the resuspended pellet were placed on pads and disposed on plates previously seeded with a defined number of conidia (see protocols to have more details). After several days at room temperature, an inhibition halo of <i>Trichoderma reesei</i>'s growth was clearly observable for the strain expressing D4E1 gene. The inhibition was even more noticeable with the strain carrying the GAFP-1 + D4E1 operon (see the photos below).</br><br />
However, no effect was detected for the strain expressing the GAFP-1 gene, supposing a synergistic effect between these two peptides.</br><br />
Regarding EcAMP and the triple-fungicides operon, no effect has been detected on the fungal growth. Several factors can explain these results: a number of post-transcriptional modifications are required to have a functional EcAMP and in addition to that, sequencing results of these constructs showed some differences with the original designed sequence.<br />
<p class="texte">Inhibition halos are not visible with supernatants, probably because of their low concentrations in the extracellular medium. <br />
Another effect was noted with the same strains expressing D4E1 and GAFP-1 + D4E1 on another fungus <i>Aspergillus brasiliansis</i>. This effect is comparable to the one previously noted with a low concentration of sulfate copper. </br><br />
</p><br />
<br />
</br><br />
<img style="width:400px; " src="https://static.igem.org/mediawiki/parts/c/c2/Resultfong.jpg"><br />
<img style="width:400px; " src="https://static.igem.org/mediawiki/parts/9/92/Results_fong_2.jpg"> <p class="legend">Figure 22: Results with transformed bacteria.</p><br />
<br />
<p class="texte"><br />
The choice of our chassis appears to be optimal as we noted that wild type <i>Bacillus subtilis</i> disturbs the hyphae growth of the fungi. Some strains of <i>Bacillus subtilis</i> (qst 713) are already used as Biofungicides for use on several minor crops to treat a variety of plant diseases and fungal pathogens.</br><br />
After this set of experiments, the strains expressing D4E1 and expressing GAFP-1 + D4E1 have shown to be the best candidates to play a major role in the fight against fungal diseases such as Canker stain. Keeping in mind our objective, <b> we decided to tests these strains in model plants</b>: <i>Nicotiana benthamiana</i> and <i>Arabidopsis thaliana</i>.</br><br />
These tests were performed in the National Institute for the Agronomic Research by experts in this domain. <br />
<br />
<br />
<p class="title2">3. <i>In planta</i> tests with SubtiTree</p><br />
<br />
<br />
<center><img style="width:215px;" img src="https://static.igem.org/mediawiki/parts/a/af/In_planta.jpg" style="margin-top:5px"/></center><br />
<p class="legend"> Figure 23: Injection of SubtiTree in a model plant</p><br />
<br />
<p class="texte"><br />
The goal of the project is to introduce the transformed bacteria in a diseased tree. So it is necessary to perform <i> in planta </i> tests to judge the fungus-killing abilities of the two strains selected after the previous set of experiments. </br><br />
SubtiTree is first inoculated in two model plants (<i>Arabidopsis thaliana</i> and <i>Nicotiana benthamiana</i>). After this step, a phytopathogenic fungus (<i>Sclerotinia sclerotiorum</i>) is placed on the leaves. </br><br />
These tests were made in association with Sylvain Raffaële and Marielle Barascud of the National Institute for the Agronomic Research laboratory. </br><br />
</p><br />
<br />
<br />
<br />
<p class="texte">Twenty-four hours after SubtiTree inoculation, no phenotypic modification of the leaves can be detected. We can conclude that our bacterium, its introduction and the fungicides production in plants do not have deleterious effects.</br><br />
Without proper treatment, the drop of the pyhtopathogenic fungus on <i>Nicotiana benthamiana</i>'s leaves causes a necrosis halo which can be measured after 40h. The lesion size and the number of inoculated sites seem to be reduced by <i>B. subtilis</i> expressing DE41 or GAFP1-D4E1, unlike with the WT bacterium. A second set of experiments is expected to be more statistically precise.</br><br></br><br />
We did not observe any significant results for <i>Arabidopsis thaliana</i> because of the use of two plants batches with different ages.</br><br />
<br />
We can therefore conclude that when SubtiTree is in plant's physiological conditions, <b> it is harmless to the plant, and that the production of fungicides is effective, reducing the leaves necrosis. </b><br />
</p><br />
<br />
<center><img style="width:860px;" img src="https://static.igem.org/mediawiki/parts/f/f1/Results_d4%2B_gafp1.jpg"></center> <p class="legend">Figure 24: Results of <i>in planta</i> test</p><br />
<br />
<br />
<p class="texte">Thanks to the diversity of anti-fungal peptides, this strategy can be adapted to different types of diseases, with different degree of specifity, etc.</p><br />
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<h2>Experimental results</h2><br />
<p> Are our modules functionnal? </p><br />
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<p style="margin:0 auto; color:#696969; width:960px; padding-top:20px; font-size:16px;"> Results&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Experimental results</p> <br />
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<p class="texte">What were the results of our experiments ? Click on these next titles to see SubtiTree abilities.</p><br />
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<p style="font-size:1.3em; margin: 0 0 30px 0;"><a href="#" onClick="ddaccordion.collapseall('technology'); return false">Collapse all</a> | <a href="#" onClick="ddaccordion.expandall('technology'); return false">Expand all</a></p><br />
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<div class="technology">Chemotaxis</div><br />
<div class="thelanguage"><br />
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<p class="texte"> We performed several tests to demonstrate the chemotaxis ability of the transformed <i>Bacillus subtilis</i> strain towards NAG and we used the wild type bacteria and glucose as a positive glucose. <br />
</p><br />
<p class="title2">1. Petri Dishes Test </p><br />
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<p class="texte"> We first tried to test chemotaxis onto Petri Dishes filled with a 0.3% agar medium. This semi-solid medium allows the bacterial motility. A paper disk containing an attractive compound is placed in the middle of the dish and cells are then loaded in the medium (see Figure 1). This protocol was taken from the <a href="https://2011.igem.org/Team:Imperial_College_London/Protocols_Chemotaxis">the Imperial College 2011 iGEM team</a>.</p><br />
<br />
<center><img SRC="https://static.igem.org/mediawiki/2014/0/05/Schema_1.png" alt="schema Figure 1" style="width:500px"></center><br />
<p class="legend">Figure 1: Schema showing how cells are filled in the medium. (A) Pipettes are used to put cells in the medium. (B) Bacteria should move to the attractive compound which diffuses.</p><br />
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<p class="texte">We did not have any result with WT <i>Bacillus subtilis</i> and glucose as attractive compound (Figure 2-A). <i>B. subtilis</i> is attracted by many other glucides and amino-acids, so we also tried to test diluted glucose in LB medium attractant (Figure 2-B).</p><br />
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<center><img SRC="https://static.igem.org/mediawiki/2014/f/ff/Fig2_AetB.png" alt="Figure 2" style="width:750px"></center><br />
<p class="legend">Figure 2: Chemotaxis test with Glucose as attractive compound (A) and Glucose added to LB medium as attractant (B).</p><br />
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<p class="texte"> We could not notice any difference between the petri dish with or without glucose. With an addition of LB medium to sugar, a large halo around the paper disk was noticeable. This halo may correspond to cells attracted by the solution, as it is not noticeable when cells are not added (data not shown). Anyway we did not have enough reproducible and reliable results to be satisfied with this test.<br> <br />
Furthermore, with the addition of LB medium, it is hard to make the distinction between the attractive effects and the simple growth resulting from random diffusion.</br><br />
We have started new tries using different protocols.</p><br />
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<p class="title2">2. Plug in Pond system<br />
</p><br />
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<p class="texte"><br />
This protocol we worked on is taken from a thesis (see references [1]). . A solution of <i>B.subtilis</i> is grown overnight so as to obtain a cell density of 8x10⁸ cells/mL. 10mL of the solution is mixed with 15mL of LB medium with 1.5 % agar kept at 45°.The final concentration of the obtained medium is 0.9% agar. Tetracycline is aded at 25µg/mL, in order to inhibit growth and to only observe the chemotaxis phenomenon. Plates are cooled and dried, before digging wells with a punch or 1mL tips. The wells are filled with attractive compounds (Figure 3). After one hour at room temperature, photos of the plates are taken and the results are analyzed.<br />
</p><br />
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<center><img SRC="https://static.igem.org/mediawiki/2014/c/cd/Fig3.png" alt="Figure 3" style="width:400px"></center><br />
<p class="legend">Figure 3: Schema showing how are made plug-in-pond tests.</p><br />
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<p class="texte"><br />
On our first try with <i>B. subtilis</i>, we made three wells per plate (Figure 4).The wells were filled with glucose at different concentrations and tetracycline was not added in one of the plates.<br />
</p><br />
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<center><img SRC="https://static.igem.org/mediawiki/2014/c/ce/Fig4.png" alt="Figure 4" style="width:400px"></center><br />
<p class="legend">Figure 4: Plates after 12h at room temperature.</p><br />
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<p class="texte"><br />
After an hour, no tangible results were obtained. It is only after 12hours that we were able to observe halos around the wells with glucose at 1M in the plates without tetracycline. Tetracycline concentration seems to be too high and inhibits any bacterial activity. Therfore, we have worked with tetracycline at 15µg/mL.<br />
We tried this protocol again with this new condition. We made two wells per plate (Figure 5), one with either Glucose or N-acetyl-glucosamine and one with LB medium. As previsously, no results were achieved after 1h, but after 12hours we could notice halos.<br />
</p><br />
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<center><img SRC="https://static.igem.org/mediawiki/2014/c/c3/Bsubtilis_result.png" alt="Figure 5" style="width:750px"></center><br />
<p class="legend">Figure 5: Chemotaxis test with <i>Bacillus subtilis</i> WT. The upper wells contain attractive compound and the lower contain medium without attractive compound. </p><br />
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<p class="texte"><br />
Results are not as clear as the first time, but we observed halos around the well with glucose at 250mM with and without tetracycline. We have then tried the same experiment with N-acetyl-glucosamine and we did not see any halo in the tested conditions. Thus we assumed that our <i>B. subtilis</i> 168 strain was not attracted to N-acetyl-glucosamine.<br />
However, the results are not clear, reliable and reproducible enough with the plug-in-pond protocol. Another testing protocol was then adopted. <br />
<br />
</p><br />
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<p class="texte"><br />
<b>References:</b></br><br />
[1] : Etude de la réponse adaptative à l'oxyde de triméthylamine et de son mécanisme de détection chez Escherichia coli et Shewanella oneidensis, 2008, Claudine Baraquet, université de la méditerranée Aix-Marseille II<br />
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<p class="title2">4. Capillary test between two tubes also called the tubes test</p><br />
<p class="texte">After the experiment of the plug-in-pond, we decided to construct a system by welding two Eppendorf tubes with a capillary thanks to an electric burner.</p><br />
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<center><img src="https://static.igem.org/mediawiki/2014/f/fb/Chemotaxis_-_eppendorf.png"></center><br />
<p class="legend">Figure 6: Photography of the first tubes system</p><br />
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<p class="texte">We tested this system with a fuchsin dye and water and we were able to observe the diffusion of fuchsin towards water. However this construction had a leakage next to the weld seam that we could not stop. <br />
Thus, we asked the help from the INSA glass blower, Patrick Chekroun. He designed two systems composed of two tubes linked by a capillary.</p><br />
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<center><img src="https://static.igem.org/mediawiki/2014/2/2b/Chemotaxis_-_tubes.png"><center><br />
<p class="legend">Figure 7: Scheme of the tubes system</p><br />
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<p class="texte">As we did previously, we tested this new system with fuchsin. This experiment was made with WT <i>Bacillus subtilis</i> and N-Acetylglucosamine.<br />
<br><br><br />
<i>NB: We could not see the diffusion from one tube to the other. We made the hypothesis that it was not visible by sight because of the small diameter of the capillary. <br />
</i><br><br />
<br><br />
The following strategy was used to avoid disturbance due to pressure and liquid movement through the capillary:<br><br />
- The first step was the addition of Wash Buffer until the capillary was full to avoid the presence of air bubbles which could lead to diffusion problems.<br><br />
- Then, the tube 2 was plugged with the thumb while another person was adding the bacterial solution of WT <i>Bacillus subtilis</i> in the tube 1. <br><br />
- The tube 1 was also plugged and only after the thumb could be removed from the tube 2. <br><br />
- In the same way, the N-Acetylglucosamine was added in the tube 2. <br><br />
- The same process was made with a xylose positive control.<br><br />
<br><br />
<i>NB: According to the article Chemotaxis towards sugars by </i>Bacillus subtilis, (George W. Ordal et al., 1979), <i>glucose and xylose have the same attractant power. We prefer a positive control instead of a negative one because we were not sure that this system was efficient.</i><br><br />
<br><br />
- The system was kept straight for 2hours. Every 40 minutes, we took a sample of each tube and spread it on an agar plate (dilution 1/1,000).</p><br />
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<center><img src="https://static.igem.org/mediawiki/2014/1/1b/Chemotaxis_-_tubes_photo.png"></center><br />
<p class="legend">Figure 8: Photography of the tubes system</p><br />
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<p class="texte">Unfortunately, the dilution was too high to detect any chemotaxis movement and the time was too short. We did not find any information in the literature.<br><br />
As we did not have the time to optimize this protocol we preferred using the protocol of the 2011 Imperial college iGEM team : the tips capillary test.</br><br />
</p><br />
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<p class="title2"> 5. Tips capillary system</p><br />
<p class="title3">First tips capillary system</p><br />
<p class="texte">This protocol comes from 2011 Imperial College iGEM team and was adapted by our team in several steps (See <a href="https://2014.igem.org/Team:Toulouse/Notebook/Protocols#select8">chemotaxis protocol</a>).<br />
<br><br />
In the first tips capillary system, we used parafilm to avoid any kind of air disturbance in the tips. The different steps are described below:<br><br />
- 15µL of each chemo-attractant was pipetted. <br><br />
- The bottom of tip with the pipette was then put on a piece of parafilm and the pipette was removed from the top of the tip.<br><br />
- The top of the tip was then sealed with a piece of parafilm. By this way, the sterility can be assured and the liquid stays inside the tip. <br><br />
- To finish, the level of the solution in the tip was marked.<br></p><br />
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<center><img src="https://static.igem.org/mediawiki/2014/9/94/Chemotaxis_-_tip.png"></center><br />
<p class="legend">Figure 9: Sealing of a tip with parafilm</p><br />
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<p class="texte">- After all the attractants were added in the tips, we put them on a green base to carry them. The whole process can be seen on Figure 10.<br><br />
- Each tip was immersed in 300 µL of a bacterial solution in the wells of an Elisa plate.<br></p><br />
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<center><img src="https://static.igem.org/mediawiki/2014/0/05/Chemotaxis_-_tip_and_support.png"></center><br />
<p class="legend">Figure 10: First tips capillary system</p><br />
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<p class="texte"><i>NB: the yellow carton was used to stabilize the system and keep it straight.</i><br><br />
<br><br />
- After one hour, the tips were removed from the bacteria solutions and the content of the tips was observed with a Thoma cell under the microscope.<br><br />
<br><br />
We had several problems with this system:<br><br />
- The liquid level decreased during the experiment and we did not have enough liquid to fill the Thoma cell. Thus, it was not possible to count.<br><br />
- The bacteria were moving and therefore, we could not proceed to a bacteria count.<br><br />
<br><br />
Regarding these observations we decided to spread the tips content on agar plates instead of using Thoma cell and microscopy.<br><br />
<p class="title3">Second tips capillary system<br />
</p><br />
<p class="texte"And then the revolution came! We found a multichannel pipette :D The same protocol was performed except that the parafilm was used to avoid the air entrance between the tips and the pipette and therefore the loss of liquid.<br></p><br />
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<center><img src="https://static.igem.org/mediawiki/2014/e/e4/Chemotaxis_-_pipette.png"></center><br />
<p class="legend">Figure 11: Second tips capillary system</p><br />
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<p class="title3">Improvement of the second tips capillary system</p><br />
<p class="texte">However this system was not optimal it is why we decided to use blu tack instead of parafilm: <br></p><br />
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<center><img src="https://static.igem.org/mediawiki/2014/4/42/Chemotaxis_-_pipette_and_blu_tack.png"></center><br />
<p class="legend">Figure 12: Improvement of the second tips capillary system</p><br />
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<p class="texte"><b>At that point, the protocol was approved and the final test could finally start! :-)</b><br><br />
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There was just one tiny problem… we did not have our optimized bacterium transformed with the chemotaxis module!!! That is why we concentrated our efforts on WT <i>Bacillus subtilis</i> strain.<br><br />
<br><br />
The main goal was to find an optimized control and to analyze the eventual chemotaxis of the WT strain. To avoid osmolality bias, we wanted to find a molecule which was non-attractant and with a similar molecular weight than the N-Acetylglucosamine (221.21 g/mol). Our first idea was to use fuchsin (Molecular weight: 337.85 g/mol).<br><br />
<br><br />
At the beginning, the experiment was conducted with only one negative contraol, the fuchsin and different NAG concentrations: 25mM, 250mM and 500mM. The tested strain was <i>Bacillus subtilis </i>168:<br><br />
<br></p><br />
<center><br />
<table align="center"><br />
<tr><td align=center><img src="https://static.igem.org/mediawiki/2014/8/8c/Chemotaxis_-_results_fuch.png"></td><br />
<td align=center><img src="https://static.igem.org/mediawiki/2014/f/fd/Chemotaxis_-_results_fuchsin.png"></td></tr><br />
<tr><td align=center><p class="legend">Figure 13: Fuchsin - negative control (dilution 1/50)</p></td><br />
<td align=center><p class="legend">Figure 14: NAG (25mM) (dilution 1/50)</p></td></tr><br />
</table></center><br><br />
<p class="texte">The average number of colonies with the negative control is 121. On the contrary, a cell layer is observed for the NAG plates with every concentration.<br><br />
<br><br />
Thus, we assumed that WT <i>Bacillus subtilis</i> was more attracted by NAG than fuchsin. Indeed we can neglect the bacterial growth because the test only lasts one hour. We also neglect diffusion and osmolality phenomena for the previous reasons. <br><br />
<br><br />
Unfortunately for us we forgot one major effect… Can you believe that fuchsin solution contains about 15% of ethanol?!!! This concentration can lead to the death of some cells which probably happened to our results.<br><br />
<br><br />
<b><p class="texte">This incredible and dramatic discovery destroyed all of our hopes about the God of chemotaxis! :-(</b><br><br />
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However, our team did not give up on synthetic biology ! :-) Indeed, after days of disappointment and no time left for lab work, we raised from ashes and tried to find another negative control.<br><br />
<br><br />
Hopefully, we managed to find a negative control: galactose (25mM). The article Chemotaxis towards sugars by <i>Bacillus subtilis</i> (<i>George W. Ordal et al., 1979</i>) proved that it was a poor attractant.<br><br />
<br><br />
We made our tests again with this new molecule and glucose (25mM) as positive control.<br></p><br />
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<center><img src="https://static.igem.org/mediawiki/2014/8/86/Chemotaxis_-_final_results.png"></center><br />
<p class="legend">Figure 15: Final results (dilution : 1/10,000)</p><br />
<p class="texte"> The miracle arrived! We managed to prove that our WT Bacillus subtilis was indeed naturally attracted to NAG</p><br />
<p class="texte"><i>NB: It was our last experiment. Unfortunately we were running out of time and we could not do much more test. We would like to do the experiment with a lower dilution and repeat it several times.</i><br><br />
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<b><p class="texte">Our results are not statistically significant however this result has been proved in literature.</p></b><br></p><br />
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<div class="technology">Binding module</div><br />
<div class="thelanguage"><br />
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<p class="title2">1. Preliminary experiments</p><br />
<p class="title3">Purpose</p><br />
<p class="texte">The first experiment deals with the culture conditions to see if <i>Bacillus subtilis</i> can resist to a low temperature and with the CBB buffer. To do that, several bacterial concentrations have been tested starting with an OD of 0.1 and diluting this solution to get estimated ODs of 0.05, 0.025, 0.01. These different <i>Bacillus subtilis</i> solutions were incubated 1 hour at 4°C with 500µL of CBB or water. Finally a cell count on Thoma cell counting chamber was performed.</p><br />
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<p class="title3">Results</p><br />
<p class="texte">The bacterial solutions could not be counted because of two main problems: the too high number of bacteria with the 0.1 OD or the too low number of bacteria with the 0.01 OD. Thus, the study is mostly focused on the intermediate values (Figure 16).<br />
<br/>First of all, a same cell concentration can be noticed with the presence of CBB or water with estimated ODs of 0.05 or 0.025. Moreover, twice less cells can be found in the lowest concentrations in bacteria comparing to the 0.05 OD concentration which is in agreement with the dilution ratio. <br />
<br/>Thus, the experimental conditions regarding the presence of CBB and the incubation temperature at 4°C do not harm the cell surviving.<br />
</p><br />
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<center><img src="https://static.igem.org/mediawiki/2014/c/ce/Graphe_binding_1.png" width="45%"></center><br />
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</br><br />
<p class="legend">Figure 16: CBB presence has no effect on bacteria. The bacterial concentration was measured regarding <span style="color:#0000FF">the presence</span> or <span style="color:#FF0000">the absence </span>of CBB for the observed OD (0.1) or estimated ODs (0.05, 0.025, 0.01).<br />
</p><br />
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<p class="title2">2. Binding test using engineered <i>B. subtilis</i></p><br />
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<p class="title3">Purpose</p><br />
<p class="texte">Transformed <i>Bacillus subtilis</i> with the binding module is able to produce a protein composed of the bacterial peptidoglycan bonding of LycT and the GbpA 4th domain of <i>Vibrio cholerae</i> allowing the chitin bonding. The synthetic bacterium is put with special beads composed of the polymer miming the fungal pathogen wall. After several washes, bacteria specifically attached to the chitin are put on plates and counted.</p><br />
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<p class="title3">Results</p><br />
<p class="texte">The first observation is that both bacterial solutions of wild type <i>Bacillus subtilis</i> and SubtiTree have the same concentration : 105 bacteria/mL (Figure 17). Even though there is no significant difference between both strains after the first wash, the second wash has a major effect since it allows 40 times more Wild Type bacteria to come off the beads. This result correlates with the number of bacteria binded to the beads for the synthetic strain with the binding module. <br />
<br/>Thus, the binding system seems to function correctly and leads to the bacterial attachment on the chitin.</p><br />
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</br><br />
<center><img src="https://static.igem.org/mediawiki/2014/e/ea/Graphe_binding_2.png" width="60%"></center><br />
</br><br />
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<p class="legend">Figure 17: Attachment of <i>Bacillus subtilis</i> with binding module to chitin. <span style="color:#0000FF">The WT bacteria</span> or <span style="color:#FF0000">the bacteria with the binding system</span> concentration has been determined during the different steps of the binding test. The stars represent a significant difference observed with a Student test with p<0.05.</p><br />
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<p class="title2">3. Microscopic observations</p><br />
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<p class="title3">Purpose</p><br />
<p class="texte">We want to observe the SubtiTree's binding on beads coated with chitin. In order to perform a 3D reconstruction showing this interaction, we use confocal laser scanning microscope. Through the use of a fluorochrome (Syto9), we can highlight the presence of bacteria on the surface of the beads (individualized by phase-contrast). A first calibration step determine the minimum threshold to remove the background noise and the natural fluorescence.</p><br />
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<p class="title3">Results</p><br />
<p class="texte">First, we note the great bacterial presence on the surface of beads coated with chitin. These images seem to highlight their interactions.</br></p><br />
<center><img src="https://static.igem.org/mediawiki/2014/archive/5/53/20141013073044!Photo_billes_microscopie.png" width="45%"></center><br />
</br><br />
<p class="legend">Figure 18: Microscopic view of bead surfaces coated with chitin</p><br />
<br />
<p class="texte">Using ImageJ software, we are able to create 3D pictures and movies of those comments.</br></p><br />
<center><img src="https://static.igem.org/mediawiki/2014/5/53/Photo_billes_microscopie.png" width="45%" style="float:left;"><iframe width="380" height="315" src="//www.youtube.com/embed/ztIHIKQr3g0" frameborder="0" allowfullscreen></iframe></center><br />
</br><br />
<p class="legend">Figure 19: A short movie of 3D bead surfaces coated with chitin</p><br />
<br />
<p class="texte">Finally we want to observe the bacteria after the second wash. When our bacterium has the binding module, results suggest a lower number of bacteria in the washing solution. SubtiTree is retained by the beads.</p><br />
<center><img src="https://static.igem.org/mediawiki/2014/9/97/Photo_lavage_microscopie.png" width="45%"></center><br />
<p class="legend">Figure 20: Microscopic view of bacteria after washing <br />
</p><br />
<br />
<p class="texte">Finally, overall results are consistent with the presence of functional binding system.</p><br />
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<div class="technology">Fungicides module</div><br />
<div class="thelanguage"><br />
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<p class="title2"> 1. Preliminary experiments</p><br />
<p class="title3">Tests with commercial peptides and controls</p><br />
<p class="texte">The first tests were accomplished with commercial GAFP-1 and D4E1 peptides at different concentrations (12.5µM, 25µM, 100µM). These tests were performed on different fungal strains sharing the same phylum with <i>Ceratocystis Platani</i>.<br />
As <i>Ceratocystis Platani</i> is pathogenic, we could not perform tests directly with this fungus.</br><br />
After several days at 30°C, the PDA (Potato Dextrose Agar) plates covered with fungus and commercial peptides were analyzed.</p></p><br />
<p class="texte">An inhibition halo was noticeable with commercial D4E1 peptide at 100µM on <i>Aspergillus brasiliensis</i>. Less bright halos were also present with lower concentrations. Concerning commercial GAFP-1, we did not notice any effect in the tested conditions.As positive control, a well-known chemical fungicide was used: the Copper Sulfate. The inhibition of the fungal growth was complete at 20mg/ml, and at 10mg/ml a darker halo appeared around the pad filled with Copper Sulfate as we can see on the figure below. This corresponds to a sporing halo in response to the stress generated by the fungicide.<br />
</p><br />
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<img style="width:450px; " src="https://static.igem.org/mediawiki/parts/a/a8/Prelim_tests_fung.jpg"><br />
<img style="width:450px; " src="https://static.igem.org/mediawiki/parts/f/f8/Controls_fung.jpg"><br />
<p class="legend"> Figure 21: Results of the preliminary tests</p><br />
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</br><br />
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<p class="texte">Given these results, we concluded that very high fungicide concentrations are required to inhibit the fungal growth. Following these tests, new conditions were adopted in order not to encourage too much fungal growth over bacterial growth. The culture medium was adjusted to fit our objective and to approximate the conditions found in the trees: a 'sap-like' medium was elaborated. The incubations were then carried at room temperature.<br />
</p><br />
<br />
<p class="title2">2. Test with SubtiTree</p><br />
<br />
<p class="texte">In order to test <i>Bacillus subtilis</i> mutants, it was essential to find the right balance between the fungal growth and the bacterial one. This condition was necessary to get a high concentration of peptides. In our genetic constructions, these peptides are designed to be exported in the extracellular medium.</br><br />
</br><br />
The transformed <i>Bacillus subtilis</i> strains grew at 37°C during 72h and were tested. After centrifugation, the supernatant and the resuspended pellet were placed on pads and disposed on plates previously seeded with a defined number of conidia (see protocols to have more details). After several days at room temperature, an inhibition halo of <i>Trichoderma reesei</i>'s growth was clearly observable for the strain expressing D4E1 gene. The inhibition was even more noticeable with the strain carrying the operon GAFP-1 + D4E1 (see the photos below).</br><br />
However, no effect was detected for the strain expressing the GAFP-1 gene, supposing a synergistic effect between these two peptides.</br><br />
Regarding EcAMP and the triple-fungicides operon, no effect has been detected on the fungal growth. Several factors can explain these results: a number of post-transcriptional modifications are required to have a functional EcAMP and in addition to that, sequencing results of these constructs showed some differences with the original designed sequence.<br />
<p class="texte">Inhibition halos are not visible with supernatants, probably because of their low concentrations in the extracellular medium. <br />
Another effect was noted with the same strains expressing D4E1 and GAFP-1 + D4E1 on another fungus <i>Aspergillus brasiliansis</i>. This effect is comparable to the one previously noted with low concentration of sulfate copper. </br><br />
</p><br />
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</br><br />
<img style="width:400px; " src="https://static.igem.org/mediawiki/parts/c/c2/Resultfong.jpg"><br />
<img style="width:400px; " src="https://static.igem.org/mediawiki/parts/9/92/Results_fong_2.jpg"> <p class="legend">Figure 22: Results with transformed bacteria.</p><br />
<br />
<p class="texte"><br />
The choice of our chassis appears to be optimal as we noted that wild type <i>Bacillus subtilis</i> disturbs the hyphae growth of the fungi. Some strains of <i>Bacillus subtilis</i> (qst 713) are already used as Biofungicides for use on several minor crops to treat a variety of plant diseases and fungal pathogens.</br><br />
After this set of experiments, the strains expressing D4E1 and expressing GAFP-1 + D4E1 have shown to be the best candidates to play a major role in the fight against fungal diseases such as Canker stain. Keeping in mind our objective, <b> we decided to tests these strains in model plants</b>: <i>Nicotiana benthamiana</i> and <i>Arabidopsis thaliana</i>.</br><br />
These tests were performed in the National Institute for the Agronomic Research by experts in this domain. <br />
<br />
<br />
<p class="title2">3. <i>In planta</i> tests with SubtiTree</p><br />
<br />
<br />
<center><img style="width:215px;" img src="https://static.igem.org/mediawiki/parts/a/af/In_planta.jpg" style="margin-top:5px"/></center><br />
<p class="legend"> Figure 23: Injection of SubtiTree in a model plant</p><br />
<br />
<p class="texte"><br />
The goal of the project is to introduce the trasnformed bacteria in a diseased tree. So it is necessary to perform <i> in planta </i> tests to judge the fungus-killing abilities of the two strains selected after the previous set of experiments. </br><br />
SubtiTree is first inoculated in two model plants (<i>Arabidopsis thaliana</i> and <i>Nicotiana benthamiana</i>). After this step, a phytopathogenic fungus (<i>Sclerotinia sclerotiorum</i>) is placed on the leaves. </br><br />
These tests were made in association with Sylvain Raffaële and Marielle Barascud of the National Institute for the Agronomic Research laboratory. </br><br />
</p><br />
<br />
<br />
<br />
<p class="texte">Twenty-four hours after SubtiTree inoculation, no phenotypic modification of the leaves can be detected. We can conclude that our bacterium, its introduction and the fungicides production in plants don't have deleterious effects.</br><br />
Without proper treatment, the drop of the pyhtopathogenic fungus on <i>Nicotiana benthamiana</i>'s leaves causes a necrosis halo which can be measured after 40h. The lesion size and the number of inoculated sites seem reduced by <i>B. subtilis</i> expressing DE41 or GAFP1-D4E1, unlike with the WT bacterium. A second set of experiments is expected to be more statistically precise.</br><br></br><br />
We did not observe any significant results for <i>Arabidopsis thaliana</i> because of the use of two plants batches with different ages.</br><br />
<br />
We can therefore conclude that when SubtiTree is in plant physiological conditions, <b> it is harmless to the plant, and that the production of fungicides is effective, reducing the leaves' necrosis </b>.<br />
</p><br />
<br />
<center><img style="width:860px;" img src="https://static.igem.org/mediawiki/parts/f/f1/Results_d4%2B_gafp1.jpg"></center> <p class="legend">Figure 24: Results of <i>in planta</i> test</p><br />
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<p class="texte">Thanks to the diversity of anti-fungal peptides, this strategy can be adapted to different types of diseases, with different degree of specifity, etc.</p><br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/Acknowledgements/SponsorsTeam:Toulouse/Acknowledgements/Sponsors2014-10-16T14:43:08Z<p>Laureen: </p>
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<p style="color:#696969; padding-top:20px; font-size:16px; float:left;"> Acknowlegdements&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Sponsors</p> <br />
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<p class="texte">Financial support is mandatory to reach the success of our project. All of our sponsors are committed to promoting the scientific and innovative experiences such as the iGEM project. We would like to thank our collaborators for their contribution to our work by giving us an outstanding overview and help. The iGEM adventure could not have been made without their support and their strong knowledge in synthetic biology.<br />
<br>Thanks to the public and private sponsors, the Toulouse iGEM team collected <B>€ 60,082.73 </B>. The encouragement was based on financial support from our schools and other partners, gifts of consumables or special sale prices on laboratory equipment.<br />
</p><br />
<br />
<p class="texte"><br />
<img style="width:180px; float:left;margin-right:10px" src="https://static.igem.org/mediawiki/2014/9/97/INSA_Toulouse_new_logo.jpg">INSA Toulouse (Institut National des Sciences Appliqu&eacute;es) is a famous engineering school allowing the specialization in 8 different subjects such as civil engineering, biochemistry but also informatics. The school hires 220 teachers and researchers and not less than 8 research laboratories.<br />
</p><br />
<br />
<p class="texte"><img style="width:200px; float:right;margin-left:10px" src="https://static.igem.org/mediawiki/2014/7/7f/Logo_UPS.jpg"><br />
L'Université Paul Sabatier is a multidisciplinary University involved in sciences and technologies, sport, gestion and communication but also health and medical studies.</p><br />
<br />
<p class="texte"><img style="width:100px; float:left;margin-right:10px" src="https://static.igem.org/mediawiki/2014/6/60/Logo_Minist%C3%A8re.gif"><br><br />
Le Ministère de l'Agriculture, de l'Agroalimentaire et de la Forêt (MAAF) was created in 1881 and implements the government's policies in agriculture, food market industries and forest.</p><br />
<br />
<br></br><br />
<br />
<p class="texte"><img style="width:100px; float:right;margin-left:10px" src="https://static.igem.org/mediawiki/2014/e/e2/Logo_VNF.jpg"><br />
Voies Navigables de France (VNF) is a French public establishment of an administrative nature created in 1991 which aims to manage the major part of the navigable waterways in France.</p><br />
<br />
<p class="texte"><img style="width:100px; float:left;margin-right:10px" src="https://static.igem.org/mediawiki/2014/5/5f/Logo_LISBP.png"><br />
The Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP) was created in 2007 and is based on the fusion of the Laboratory of Biotechnology and Bioprocesses and the Laboratory of Engineering in Environmental Processes. The LISBP is mostly specialized in innovative technics regarding living and process sciences such as microbiology, biocatalysis and separation technics.</p><br />
<br />
<p class="texte"><img style="width:120px; float:right;margin-left:10px" src="https://static.igem.org/mediawiki/2014/4/4a/Logo_TWB.png"><br />
Toulouse White Biotechnology (TWB) is a French infrastructure developing new technics of sustainable production by enhancing the use of renewable carbon source. TWB has three main missions: promoting the white biotechnologies, improving the scientific innovation, binding the fundamental research and the industrial world. White biotechnologies are the used of industrial goals, including the use of cells and enzymes to create industrial products.</p><br />
<br />
<br />
<p class="texte"><img style="width:130px; float:left;margin-right:10px" src="https://static.igem.org/mediawiki/2014/0/05/Logo_Adisseo.jpg"><br />
Adisseo is a subsidiary of Bluestar Company and a worldwide expert in animal nutrition. This branch has a major role in the improvement of food chain while preserving animals, mankind and environment.</p><br />
<br />
<br />
<p class="texte"><img style="width:100px; float:right;margin-left:10px" src="https://static.igem.org/mediawiki/2014/6/61/Logo_Genotoul.png"><br />
The Genopole of Toulouse (GenoToul) was created in 1999 to develop genomes studies, bioinformatics and to promote the innovation by encouraging interdisciplinary projects. GenoToul welcomes around fifty projects each year regarding many subjects such as healthcare, agronomy, biotechnologies, ecology and environment. </p><br />
<br />
<p class="texte"><img style="width:80px; float:left;margin-right:10px" src="https://static.igem.org/mediawiki/2014/0/0d/Logo_COMUE.jpeg"><br />
The Communauté d'Universités et d'Etablissement de Toulouse (COMUE) includes 14 establishments such as universities, engineering schools, specialised schools. It aims to position the Toulouse universitary site at the best european and international level. </p><br />
<br />
<p class="texte"><img style="width:90px; float:right;margin-left:10px" src="https://static.igem.org/mediawiki/2014/5/5d/Logo_LabexTulip.jpg"><br />
Labex Tulip is a laboratory composed of five units specialized in agrobiosciences and environment. It reprents a potential for research with approximately 400 people which belongs to many different laboratories. Labex Tulip is focused on the interactions between organisms and the community. The main goal is to enhance the research studies and the attractivity of its laboratories.</p><br />
<br />
<p class="texte"><img style="width:100px; float:left;margin-right:10px" src="https://static.igem.org/mediawiki/2014/5/54/Logo_CROUS.jpg"><br />
The Centre Régional des Oeuvres Universitaires et Scolaires (CROUS) is a public facility controlled by the Ministery of Higher Education and Research. The main goal is to promote the good living conditions of students. The CROUS can provide help in many different actions: direct help through financial fundings, undirect help with housing and catering. </p><br />
<br />
<p class="texte"><img style="width:80px; float:right;margin-left:10px" src="https://static.igem.org/mediawiki/2014/8/88/Herault.jpg"><br></br>The Hérault Département is in the Languedoc-Roussillon region with more than 1,000,000 inhabitants.</p><br />
<br />
<br />
<p class="texte"><img style="width:100px; float:left;margin-right:10px" src="https://static.igem.org/mediawiki/2014/0/02/Castelnaudary.jpg">Castelnaudary is a 12,000 inhabitant’s city located in Aude department where Pierre Paul Riquet constructed a pond named Le Grand Bassin.</p><br />
<br />
<p class="texte"><img style="width:70px;float:right;margin-left:10px" src="https://static.igem.org/mediawiki/2014/4/48/Labastide_d%27Anjou.png"> Labastide-d’Anjou is a 1,500 inhabitants’ village located in the Aude department in Languedoc-Roussillon. This fortified town is situated nearby the Canal du Midi.</p><br />
<br />
<br />
<p class="texte"><img style="width:100px; float:left;margin-right:10px " src="https://static.igem.org/mediawiki/2014/a/a6/Logo_montr%C3%A9al.png"> <br></br>Montréal is a 2,000 inhabitants’ village located next to the Montagne noire, in Aude department in Languedoc-Roussillon.</p><br />
<br />
<br />
<p class="texte"><img style="width:80px; float:right;margin-left:10px" src="https://static.igem.org/mediawiki/2014/0/0f/Sall%C3%A8les_d%27Aude.jpg"> <br></br> Sallèles d’Aude is a 3,000 village situated in Aude department in the Languedoc-Roussillon region where the “Canal de jonction” runs through which joins the Canal du Midi.</p><br />
<br />
<br />
<p class="texte"><img style="width:80px;float:left;margin-right:10px" src="https://static.igem.org/mediawiki/2014/d/db/600px-Blason_ville_fr_Ventenac-en-Minervois_%28Aude%29.svg.png"> <br></br>Ventenac en Minervois is 600 inhabitants’ village situated in Aude in the Languedoc-Roussillon region where the Canal du Midi passes through.</p><br />
<br />
<br />
<p class="texte"><img style="width:130px; float:right;margin-left:10px" src="https://static.igem.org/mediawiki/2014/2/26/Logo_ThermoFisher.png"><br />
Thermo Fisher Scientific is one of the leaders in serving science by offering a wide range of innovative technologies and support to make the world cleaner and safer. The company serves pharmaceutical, biotechnology companies as well as hospitals, clinical laboratories.</p><br />
<br />
<p class="texte"><img style="width:100px; float:left;margin-right:10px" src="https://static.igem.org/mediawiki/2014/9/91/Qiagen_logo.JPG"><br />
Qiagen was created in 1984 and is nowadays a leader in innovative market and technology by creating assay technologies that can enable the access to content from any biological sample. The main purpose is to help achieving success and major breakthroughs in life sciences. Qiagen offers a broad range of 500 products for over 500, 000 customers worldwide.</p><br />
<br />
<p class="texte"><img style="width:130px; float:right;margin-left:10px" src="https://static.igem.org/mediawiki/2014/9/9f/Logo_NEB.jpg"><br />
New Englands Biolabs (NEB) is the leading industry in discovery and production of enzymes and the offer of recombinant and native enzymes since 1970. NEB carried on expanding its product in PCR fields but also gene expression, RNA and cellular analysis. The company is mostly focused on promoting new technologies on the market.</p><br />
<br />
<p class="texte"><img style="width:150px; float:left;margin-right:10px" src="https://static.igem.org/mediawiki/2014/6/60/Eurofins_new.jpg"><br />
Eurofins Scientific is a French company created in 1987 and mostly specialized in bioanalysis with more than 100,000 analysis methods and technologies. This industry has an international network of 190 laboratories in 36 different countries with 15 000 employees and has many fields of specialization: food market, environment, pharmaceutical and medical industries, agroscience… Eurofins Scientific contributes to the well-being and the health of everyone by giving high-quality policy advice and analysis services.</p><br />
<br />
<p class="texte">If you are interested in any kind of partnership with the Toulouse iGEM team 2014, please contact us at <a href="mailto:adresse@serveur.extension">igemtoulouse2014@gmail.com</a><br />
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{{:Team:Toulouse/template/footer}}</div>Laureenhttp://2014.igem.org/Team:Toulouse/Team/Fun_factsTeam:Toulouse/Team/Fun facts2014-10-16T14:41:54Z<p>Laureen: </p>
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<p style="color:#696969; padding-top:20px; font-size:16px; float:left;"> Team&nbsp;&nbsp;&nbsp;>&nbsp;&nbsp;&nbsp;Fun Facts</p> <br />
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<p class="title2">Have you ever tried to estimate the kilometers travelled by your team during this summer?</p><br />
<p class="texte"><br />
According to our calculations, one member walks approximately 4 kilometers per day all around the <br />
laboratory. This represents 5,544 kilometers for the whole team during the 126 days of this epic <br />
adventure. <br />
What does that mean exactly? Simply that we could walk, cycle, swim or whatever you want until <br />
Kazakhstan, Russia, Kenya… We could even have reached the USA but we decided to stay in the lab <br />
and take the plane to go to Boston!</p><br />
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<div style="float:right; width:500px;"><br />
<img src="https://static.igem.org/mediawiki/2014/5/52/Interview.jpg" style="margin-top:5px; margin-left: 62px; width:450px" /><br />
</div> <br />
<br />
<p class="title2">What do trees lining the “Canal du Midi” think about SubtiTree?</p><br />
<p class="texte"><br />
According to our homemade impartial survey, 94% of the questioned plane trees approve our project and are interested in <br />
serving as guinea pigs for our new bacterial medicine. This percentage represents 41,580 trees which <br />
are also gathered in the association called: “Happy tree friends“.</p><br />
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<div style="float:left; width:500px; margin-top:50px;"><br />
<img src="https://static.igem.org/mediawiki/2014/a/ab/Multidisciplinary_yes_we_are.jpg" style="margin-top:5px; width:450px" /><br />
</div> <br />
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<br><br />
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<br> <br />
<p class="title2">Multidisciplinary…Yes we are!</p><br />
<p class="texte">Housework in our laboratory became necessary when most of people were in vacations except us. <br />
But do you know the novelty this year in our team? Times are changing because now men are <br />
cleaning! ;-)</p><br />
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<br><br />
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<div style="float:right; width:500px; margin-top:50px;"><br />
<img src="https://static.igem.org/mediawiki/2014/e/e2/World_cup.jpg" style="margin-top:5px; width:450px; margin-left:62px;" /><br />
</div> <br />
<br> <br> <br> <br> <br> <br><br />
<p class="title2">The unmissable event this summer: The 2014 football World Cup !</p><br />
<p class="texte">From 06/12/14 to 07/13/14, the Toulouse iGEM Team was cheering on the French team. During the <br />
games of the French team, our group was juggling with wet lab and large screen projections! Despite <br />
our support, the French team did not win the World Cup. However, we have not said our last world <br />
yet: let’s see what will happen in 2018… ;-)</p><br />
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<div class="clear"></div><br />
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<div style="margin-top:50px; text-align:center;"><br />
<p class="title2">Have you ever forgetten a culture tube or a petri dish?</p><br />
<p class="texte" style="text-align:center;">Never? Let us show you what happens in that case!</p><br />
</div><br />
<center><br />
<table><br />
<tr><td><img src="https://static.igem.org/mediawiki/2014/f/fa/Old_tube1.png" width="160px"></td><br />
<td><img src="https://static.igem.org/mediawiki/2014/1/1b/Old_tube2.JPG" width="400px"></td></tr><br />
<tr><td><img src="https://static.igem.org/mediawiki/2014/d/d4/Old_petri1.png" width="370px"></td><br />
<td><img src="https://static.igem.org/mediawiki/2014/d/d7/Old_petri2.JPG" width="400px"></td></tr><br />
</table><br />
</center><br />
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<div style="text-align:center; width:760px; margin:0 auto; margin-top:15px; border-top:1px solid #555; padding-top:60px;"><br />
<p class="title2" style="padding-bottom:15px;">To finish this part, let’s do the official Awards Ceremony of the 2014 Toulouse iGEM Team!</p><br />
<ul><br />
<li class="tree"><p class="texte">The Geek Award: The three nominees are Laureen, Manon, Florie. And the winner is… <b>Florie</b> <br />
who spent the longest time in front of her laptop for the modeling part!</p></li><br />
<li class="tree"><p class="texte">The latest survivor of weekly meetings: The four nominees are Fanny, Emeline, Diane, Pierre. <br />
And the winner is… <b>Diane</b> who stayed up until 3am because she was skyping from South <br />
Korea!</p></li><br />
<li class="tree"><p class="texte">The worst singer award: The two nominees are Abdel, Pierre. And the winner is… <b>Abdel</b> who <br />
spent the whole day singing badly in the lab!</p></li><br />
<li class="tree"><p class="texte">The dancer award: The three nominees are Florie, Camille, Diane. And the winner is… <b>Camille</b><br />
who did the famous Plasmid Dance!</p></li><br />
<li class="tree"><p class="texte">The “hello you” award: No nominees because the only winner is… <b>Pierre</b> who was saying <br />
“Hello you” each time he met someone!</p></li><br />
<li class="tree"><p class="texte">The most tired award: The three nominees are Laureen, Manon, Aurélie. And the winner is... <br />
<b>Manon</b> but we still do not know why!</p></li><br />
<li class="tree"><p class="texte">The misplaced ideas award: The four nominees are Laureen, Camille, Mathieu, Pierre. And <br />
the winner is… <b>Mathieu</b> but you do not want to know why!</p></li><br />
<li class="tree"><p class="texte">The perseverance award: The three nominees are Emeline, Diane, Abdel. And the winner is... <br />
<b>Emeline</b> who succeeded a cloning after twelve trials!</p></li><br />
<li class="tree"><p class="texte">The drawing award: The three nominees are Florie, Fanny, Manon. And the winner is… <b>Fanny</b> <br />
who drew our first SubtiTree logo!</p></li><br />
<li class="tree"><p class="texte">The phone-call award: The two nominees are Laureen, Pierre. And the winner is… <b>Laureen</b><br />
who was our lab secretary!</p></li><br />
<li class="tree"><p class="texte">The biggest blunder in the lab award: The three nominees are Aurélie, Fanny, Abdel. And the <br />
winner is… <b>Aurélie</b> who poured an agarose gel without gel tray!</p></li><br />
<li class="tree"><p class="texte">And last but not least ... The Best Nervous breakdown Award goes to ... <b>Our deep freezer</b>! The whole team is grateful for its hard work during a hot summer!</li></p><br />
</ul><br />
</div><br />
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