http://2014.igem.org/wiki/index.php?title=Special:Contributions&feed=atom&limit=250&target=Alquiru&year=&month=2014.igem.org - User contributions [en]2024-03-29T11:26:35ZFrom 2014.igem.orgMediaWiki 1.16.5http://2014.igem.org/Team:Valencia_UPV/Team/AttributionsTeam:Valencia UPV/Team/Attributions2015-02-19T16:49:04Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Team</a> > <a>Attributions</a></h3></p><br/><br />
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<div align="center"><span class="coda"><roja>A</roja>ttributions</span> </div><br/><br/><br />
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<h3>Attributions in a flash</h3><br/><br />
<p><a name="Alba" class="encabezado">Alba Rubert:</a> Pheromone Analysis, Pheromone-Insect Interactions, BioBrick preparation, Policy and Practices.</p><br/><br />
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<p><a name="Alejandra" class="encabezado">Alejandra González:</a> Pheromone Diffusion modeling, Pheromone Production modeling, Policy and Practices.</p><br/><br />
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<p><a name="Alfredo" class="encabezado">Alfredo Quijano:</a> Pheromone Release, Switch, Pheromone Analysis, Pheromone-Insect Interactions, Policy and Practices.</p><br/><br />
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<p><a name="Ivan" class="encabezado">Ivan Llopis:</a> Pheromone Production modeling, Wiki, Policy and Practices.</p><br/><br />
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<p><a name="Jose" class="encabezado">José Gavaldá:</a> Pheromone Biosynthesis design, Pheromone Analysis, Policy and Practices</p><br/><br />
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<p><a name="Lucia" class="encabezado">Lucía Estellés:</a> Pheromone Biosynthesis design, Biosafety, BioBrick preparation, Policy and Practices.</p><br/><br/><br/><br />
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<h3>Attributions (Extended Version)</h3><br/><br />
<p><br />
<strong>Project: the idea.</strong><br />
We came to The Sexy Plant project after many brainstorming meetings including all the team. It would be impossible to define authors here: we all contributed to the design of the project.<br />
</p><br/><br />
<p><br />
<strong>Pheromone Biosynthesis.</strong><br />
Design of the biosynthesis pathway fell on <strong>Jose Gavaldá</strong> and <strong>Lucía Estellés</strong>. Pheromone expression analysis by GC-MS was carried out by <strong>Alba Rubert</strong>, <strong>Alfredo Quijano</strong>, <strong>Lucía Estellés</strong> and <strong>Jose Gavaldá</strong>. Pheromone-insect interaction analysis by means of wind tunnel and EAG was made by <strong>Alba Rubert</strong> and <strong>Alfredo Quijano</strong>.<br />
</p><br />
<p><br/><br />
<strong>Pheromone Release and Switch.</strong><br />
<strong>Alfredo Quijano</strong> was the responsible for the design and expression analysis of the trichome specific promoter. Copper inducible switch was designed by <strong>Alfredo Quijano</strong>. <br />
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<strong>Modeling.</strong><br />
<strong>Ivan Llopis</strong>, also supported by <strong>Alejandra González</strong>, worked in the FBA modeling to try to obtain estimation of the pheromone production and to try to optimize it. <strong>Alejandra González</strong> was in charge of the pheromone diffusion and moth behavior modeling to help decide where and how much sexy plants we should put in the field.<br />
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<strong>Biosafety.</strong><br />
Biosafety module was designed by <strong>Lucía Estellés</strong>, and Lucía was also the leader when fulfilling safety requirements: About our lab form (assisted by Alba, Alfredo and Jose), Check-in and Safety form.<br />
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<p><br />
<strong>Policy & Practices.</strong><br />
<strong>Lucía Estellés</strong>, <strong>Jose Gavaldá</strong>, <strong>Alejandra González</strong>, <strong>Alba Rubert</strong> and <strong>Alfredo Quijano</strong> dealed with the organization and execution of interviews with some stakeholders involved in the project. All the students in the team participated in the presentation of the Sexy Plant project to relevant members of the UPV, CSIC (Spanish Research Council) and Bayer CropScience business chair. <strong>Alba Rubert</strong>, <strong>Alfredo Quijano</strong>, <strong>Alejandra González</strong> and <strong>Ivan Llopis</strong> participated in the Generación Espontánea event. Also all the students organized and carry out the Summer School courses. And Lucía, Jose, Alba and Alfredo organized and participated in the lipdub.<br />
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<p><br />
<strong>BioBricks.</strong><br />
<strong>Lucía Estellés</strong>, helped by <strong>Alba Rubert</strong>, prepared the BioBricks for sending them to the Registry and documented the parts. <strong>Lucía</strong> was the designer of the Omega undercover part used to translate from GoldenBraid 2.0 to BioBricks standard.<br />
</p><br/><br />
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<strong>Interlab Study.</strong><br />
<strong>Lucía Estellés</strong> performed the measurement interlab study.<br />
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<strong>Art and Design.</strong><br />
<strong>Alfredo Quijano</strong> was the major designer of the images used at the wiki, the presentation and the poster.<br />
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<strong>Wiki. </strong><br />
Performed by <strong>Ivan Llopis</strong>.<br />
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<strong>Poster.</strong><br />
<strong>Ivan Llopis</strong> worked in the poster design, all contributed with the concepts.<br />
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<strong>Talk.</strong> <strong>Lucía Estelles, Alfredo Quijano</strong> and <strong>Alejandra González.</strong> Also <strong>Alba Rubert</strong> performed Lucía's part when she was absent. All the team participated in the layout and conceptual flow. <br />
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<h3>Supervision</h3><br/><br />
<p><br />
All the Valencia UPV supervisors and advisors performed outstanding jobs. Their implication in the project is as follows:<br />
</p><br/><br />
<p><br />
<strong>Wetlab work</strong>. It was supervised by <strong>Estefanía Huet</strong> and <strong>Marta Vázquez</strong>. Always under the guidance and supervision of the wetlab director: <strong>Prof. Diego Orzáez.</strong><br />
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<p><br />
<strong>Modeling.</strong> FBA analysis was closely supervised by <strong>Yadira Boada</strong>, with the collaboration of <strong>Gabriel Bosque</strong> and <strong>Maria Siurana</strong>. Under the guidance of<strong> Profs. Javier Urchueguía and Jesús Picó. </strong>. Modeling of the Pheromone Diffusion and Moths response was supervised by <strong>Alejandro Vignoni</strong>. All under the guidance of<strong> Profs. Jesús Picó and Alberto Conejero. </strong><br />
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<strong>Wiki.</strong> Supervised by <strong>Victor Nina</strong> and <strong>Alejandro Vignoni</strong>.<br />
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<p><br />
<strong>Communication and logistics.</strong> <strong>Javier Urchieguía, Alberto Conejero, Maria Siurana, David Fuente</strong> and <strong>Yadira Boada</strong>.<br />
</p><br/><br />
<p><br />
<strong>Poster design and layout</strong> was supervised by<strong> Yadira Boada</strong>.<br />
</p><br/><br/><br/><br />
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<h3>Acknowledgements</h3><br/><br />
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<p>We thank the people who have helped in the project. We sincerely appreciate your contributions because <i>The Sexy Plant</i> project would not have been possible without your invaluable assistance and your enthusiasm: </p><br/><br />
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<ul class="method"><br />
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<li>Asun Fernández del Carmen, Institute for Plant Molecular and Cell Biology (IBMCP, CSIC-UPV): invaluable aid in everything. </li><br />
<li>Jose Luis Rambla Nebot, Institute for Plant Molecular and Cell Biology (IBMCP, CSIC-UPV): aid and advising in sample analysis by CG-MS and other technical advice. </li><br />
<li>Jaime Primo Millo, Ismael Navarro Fuertes, Vicente Navarro Llopis, Sandra Vacas González, Centre for Agricultural Chemical Ecology (CEQA - UPV): technical advice in project approach, insect management, electroantennography, wind tunnel, plant diffusion analysis, facility and technical details provision. </li><br />
<li>Christine Bäuerl, Institute of Automation and Industrial Computing ai2 - UPV: technical advice in the laboratory. </li><br />
<li>Jesús Muñoz Bertomeu, Institute for Plant Molecular and Cellular Biology (IBMCP, CSIC-UPV): <i>Candida tropicalis species</i> and assistance. </li><br />
<li>Lynne Yenush and Mª Carmen Marqués Romero, Institute for Plant Molecular and Cell Biology (IBMCP, CSIC-UPV): <i>Saccharomyces cerevisiae species</i>, genomic extraction protocols and reagents. </li><br />
<li>Alejandro Sarrión Perdigones, Baylor College of Medicine (Houston, Texas): technical advice and RFC development. </li><br />
<li>Matilde Eizaguirre Altuna, Department of Crop and Forest Sciences, University of Lleida (Lleida, Spain): <i>S. nonagrioides species</i> and useful information about moth handling. </li><br />
<li>NRP-UEA-Norwich team: MoFlippers and chromoproteins.. </li><br />
<li>IBMCP (CSIC-UPV) staff: Lipdub participation, hosting us at the Institute. </li><br />
<li>Silvia Michael: Logo design. </li><br />
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</ul><br/><br />
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<p>We would like to offer our special thanks to the following institutions for their support in the development of <i>The Sexy Plant Project</i>: </p><br/><br />
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<li>Universitat Politècnica de València (UPV) and Vice-rectorate for Student and University Extension, </li><br />
<li>Council of Valencia, </li><br />
<li>Bayer CropScience business chair, </li><br />
<li>Technical School of Industrial Engineering (ETSII - UPV), </li><br />
<li>Technical School of Agricultural Engineering (ETSIAMN - UPV) </li><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Team/Students/Alfredo_QuijanoTeam:Valencia UPV/Team/Students/Alfredo Quijano2015-02-19T16:48:01Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Team</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students">Students</a> > <a>Alfredo Quijano Rubio</a></h3></p><br />
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<img alt="Profile_Alfredo" title="Alfredo Quijano Rubio" src="https://static.igem.org/mediawiki/2014/5/52/Profile_Alfredo.jpg" class="img-profile-desc"></img><br />
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<h4 class="title-desc">Alfredo Quijano Rubio</h4><br/><br />
<p>He’s currently studying 4th year of Biotechnology bachelor’s degree in the UPV “Universitat Politècnica de València”. He has always been keen on Synthetic Biology and once iGEM Giant Jamboree 2014 has finished he plans to keep working on Plant Synthetic Biology and start his Final Degree Project developing new plant genetic switches.</p><br/> <br />
<p>In his leisure time, he enjoys woodworking and sports.<br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Team/Students/Alfredo_QuijanoTeam:Valencia UPV/Team/Students/Alfredo Quijano2015-02-19T16:45:24Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Team</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students">Students</a> > <a>Alfredo Quijano Rubio</a></h3></p><br />
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<img alt="Profile_Alfredo" title="Alfredo Quijano Rubio" src="https://static.igem.org/mediawiki/2014/5/52/Profile_Alfredo.jpg" class="img-profile-desc"></img><br />
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<h4 class="title-desc">Alfredo Quijano Rubio</h4><br/><br />
<p>He’s currently studying 4th year of Biotechnology bachelor’s degree in the UPV “Universitat Politècnica de València”. He has always been keen on Synthetic Biology and once iGEM Giant Jamboree 2014 has finished he plans to keep working on Plant Synthetic Biology and start his Final Degree Project developing new plant genetic switches.</p><br/> <br />
<p>In his leisure time, he enjoys woodworking and sports. <br/><br/><br />
ROSANA GALLENT, JULIA HERNÁNDEZ y GUILLEM FERRERES SON UNOS PERSONAJES<br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/AchievementsTeam:Valencia UPV/Achievements2014-10-18T03:52:15Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Policy and Practices</a> > <a>Achievements</a></h3></p><br/><br />
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<div align="center"><span class="coda"><roja>A</roja>chievements</span> </div><br/><br/><br />
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<p>As it can bee observed surfing our wiki, at the end of this long road we have accomplished many positive results:</p><br />
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<ul class="method"><br />
<li><a class="black-bold">A plant able to produce three insect sexual pheromones from moths to produce insects mating disruption, the Sexy Plant.:</a> <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis" class="normal-link-page">Results: Pheromone analysis</a><br />
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<ul><br />
<li>Obtained pheromones being among the most abundant plant organic volatile compounds.<span class="red-bold"> (Z)-11-hexadecen-1-ol</span> is certainly the most abundant one.</li><br />
<li>Successful and functional assembly of each of the pheromone biosynthetic genes with the plant constitutive promoter P35S. Also multigenic assembly of all three transcription units in a single plasmid. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#biosyn" class="normal-link-page"> Results: Constructs-Biosynthesis</a></li><br />
<li>Proof of our produced pheromones-insect interaction. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/eag" class="normal-link-page">Results: Electroantennography</a></li><br />
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</li><br />
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<ul class="method"><br />
<li><a class="black-bold">A broad understanding of plant metabolism, by adapting and exploitation of a genome-scale model of Arabidopsis thaliana primary metabolism able to help us theoretically optimize the pheromone production </a> <a href="https://2014.igem.org/Team:Valencia_UPV/Modeling/fba" class="normal-link-page"> Modeling: Pheromone Production</a><br />
<ul><br />
<li>Adapting the AraGEM genome-scale model to our pheromone production pathway by branching the flux of our precursor metabolite Palmitic acid (16:0).</li><br />
<li>Identifying the principal: i) cell compartments and scenarios of the plant metabolism, ii) flux bounds and constraints for each scenario, and iii) the interactions between chemical reactions and substrates related to our pheromone. </li><br />
<li>Exploring genetic conditions that could improve our pheromone production by Single gene Knock-out analysis. </li><br />
<li>Obtaining optimal condition for the coupled yield of pheromone and biomass production as a function of photons consumption.</li> <br />
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</li><br />
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<ul class="method"><br />
<li><a class="black-bold">A plant potentially able to release the produced pheromones into the environment.</a><br />
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<ul><br />
<li>Successful cloning of a trichome-specific promoter (PCPS2) from the genome of <i>N. tabacum</i> and subsequent assembly with GFP as a reporter. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#phero" class="normal-link-page">Results: Constructs-Pheromone release</a></li><br />
<li>Proof of the specificity of this promoter by GFP fluorescence detection. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/trichome_expression" class="normal-link-page">Results: Trichome-specific expression</a></li><br />
<li>Assembly of each gene of the pheromones production pathway with PCPS2 promoter and multigenic assembly with all three transcription units in a single plasmid. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#phero" class="normal-link-page">Results: Constructs-Pheromone release</a></a></li><br />
</ul><br />
</li><br />
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<ul class="method"><br />
<li><a class="black-bold">A Simulation Environment for the pheromone diffusion and moth response behaviour to help us decide where and how much sexy plants we should put in the field. </a> <a href="https://2014.igem.org/Team:Valencia_UPV/Modeling/diffusion" class="normal-link-page"> Modeling: Pheromone Diffusion and Moths Response</a><br />
<ul><br />
<li>Implementing an approximation of the diffusion process by the heat diffusion equation and its numerical solution.</li><br />
<li>Incorporating an approximation of the moth response behavior including female pheromone release into the environment, and male pheromone concentration sensitivity (that allows the male to follow the trace of females).</li><br />
<li>Adding our sexy plants that produce and release pheromone triggering <i>mating disruption</i>.</li><br />
<li>Designing a simulation platform available to community that is useful for exploration of parameters and scenarios related with chemical ecology models.</li><br />
</ul><br />
</li><br />
</p><br />
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<ul class="method"><br />
<li><a class="black-bold">A genetic switch able to control gene expression ready to be implemented in the plant.</a><br />
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<ul><br />
<li>Cloning of the coding sequence from the CUP2 transcription factor from S cerevisiae and assembly with the CaMV constitutive promoter P35S. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#switch" class="normal-link-page">Results: Construct-Switch</a></li><br />
<li>Creation of the Cupper-responsive chimeric promoter and assembly with Firefly luciferase gene as a reporter, P19 as a gene silencing suppressor, and Renilla luciferase as control for Luciferase expression assay. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#switch" class="normal-link-page">Results: Construct-Switch</a></li><br />
<li>Multigenic assembly comprising the CUP2 transcriptional unit with the chimeric promoter and reporter gene assembly. <br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#switch" class="normal-link-page">Results: Construct-Switch</a></li><br />
</ul><br />
</li><br />
</p><br/><br/><br />
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<ul class="method"><br />
<li><a class="black-bold">A sterile and dark purple plant safe for living beings and the environment.<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety" class="normal-link-page">Biosafety</a><br />
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<ul><br />
<li>Multigenic assembly of two biosafety devices, comprising the Barnase (male-sterility) with one chromoprotein in each device, AmilCP or AmilGFP (identity preservation). <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#biosafe" class="normal-link-page"> Results: Constructs-Biosafety</a></li><br />
<li>Purple plant to preserve its identity expressing SlANT1 and SlJAF13 transcription factors. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/biosafety" class="normal-link-page"> Results: Biosafety</a></li><br />
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</ul><br />
</li><br />
</p><br/><br/><br />
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<p><br />
<a class="black-bold">Diffusion and communication of the project with involved experts and stakeholders.<a href="https://2014.igem.org/Team:Valencia_UPV/policy/overview" class="normal-link-page"> Policy and Practices</a></a> <br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Medal_requierementTeam:Valencia UPV/Medal requierement2014-10-18T03:49:19Z<p>Alquiru: </p>
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<h3 class="hook" align="left"><a>Achievements</a> > <a>Medal Requirements</a></h3></br><br />
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<div align="center"><span class="coda"><roja>M</roja>edal <roja>R</roja>equirements</span> </div><br/><br />
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<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/2/2b/VUPVEdiGEM_bronze_medal.png" alt="bronze_medal"></img><span class="background_medal"><b>Bronze Medal Requirements</b></span><br/><br />
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<ul class="medals-tick" id="bronze"><br />
<li><a class="black-bold">Team registration</a></li><br/><br />
<li><a class="black-bold">Team Wiki</a></li><br/><br />
<li><a class="black-bold">Complete <a class="normal-link-page"href=" https://igem.org/2014_Judging_Form?id=1554">Judging Form</a> </a></li><br/><br />
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<li><b>Present a poster and a talk at the iGEM Jamboree</b></li><br />
Poster ready to go!<br/><br/><br />
<li><b>The description of each project must clearly attribute work done by the students and distinguish it from work done by others, including host labs, advisors, instructors, sponsors, professional website designers, artists, and commercial services. </b></li><br />
<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Team/Attributions">See our Attributions here!</a><br/><br/><br />
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<li><b> Document and submit new standard BioBrick Parts or Device to the iGEM Registry .<br/> <br />
<a class="normal-link-page"href="https://2014.igem.org/Team:Valencia_UPV/VUPV_Parts">See our Parts here!</a></b></li><br/> <br />
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<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/5/5d/VUPVEdiGEM_silver_medal.png" alt="silver_medal"></img><span class="background_medal"><b>Silver Medal Requirements</b></span><br/><br />
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<ul class="medals-tick" id="silver"><br />
<li><b>Experimentally validate that your new BioBrick Parts or Devices work as expected:</b><br/><br />
We can proudly assure that the following BioBricks work perfectly:<br />
<ul><br />
<li>BBa_K1554001, BBa_K1554002, BBa_K1554003 are the three enzymes that constitute the pheromone synthesis pathway. They proved to work and provided the expected results (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis">Results: Pheromone Analysis</a>).</li><br />
<li>BBa_K1554006 allows an efficient conversion from GoldenBraid 2.0 standard parts to BioBrick standard parts (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/parts_construction">Parts Contruction</a>).</li><br />
</ul></li><br/><br />
<li><b> Document the characterization of these parts in the “Main Page” section of that Part’s/Device’s Registry entry:</b><br/><br />
Check it! <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554001">BBa_K1554001</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554002">BBa_K1554002</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554003">BBa_K1554003</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554006">BBa_K1554006</a></li><br/><br />
<li><b>Submit this new part to the iGEM Parts <br/><br />
<a class="normal-link-page" href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2014&group=Valencia_UPV">Registry</a></b></li><br/><br />
<li><b>Articulate questions encountered by your team, and describe how you considered them within your project</b></li><br />
<li style="list-style: none;">To tune our project and ensure its outcome is in line with societal expectations, we engaged a wide range of social actors and stakeholders throughout the whole of it. Visit our <a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/policy/overview">Policy and Practices</a> section.</li><br/><br />
</ul><br />
<br/><br/><br />
<br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/8/87/VUPVEdiGEM_gold_medal.png" alt="gold_medal"></img><span class="background_medal"><b>Gold Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="gold"><br />
<br />
<li><b>Improve the function OR characterization of an existing BioBrick Part or Device and enter this information in the Registry:</b></li><br />
<li style="list-style: none;">In our biosafety module we have used the Barnase (BBa_J72173), AmilGFP chromoprotein (BBa_K1467202) and AmilCP chromoprotein (BBa_K1467201) from the parts Registry, and tested them in <i>Nicotiana benthamiana </i>. We also have used Golden Gate Module Flippers (BBa_K1467100, BBa_K1467200) to translated our GoldenBraid 2.0 standard parts to BioBricks’.</li><br/><br />
<br />
<li><b>Help any registered iGEM team from another school or institution by, for example, characterizing a part, debugging a construct, or modeling or simulating their system</b></li><br />
<li style="list-style: none;">We developed a tight collaboration with the <a href="https://2014.igem.org/Team:NRP-UEA-Norwich" class="normal-link-page">NRP-UEA-Norwich</a> team. In collaboration, our team developed two biosafety devices thanks to two parts provided by them. Both devices were submitted to the iGEM Registry BBa_K1554004 and BBa_K1554005. In addition, the NRP-UEA-Norwich also provided us with their <a href="https://2014.igem.org/Team:NRP-UEA-Norwich/Project_Mo-Flipper" class="normal-link-page">Mo-Flippers</a>. We tested them in GoldenBraid standard parts and they worked perfectly.</li><br/><br />
</br></br></br><br/><br />
<br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/interlab"><strong>&larr; Go to Interlab Study</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Achievements"><strong>Go to Achievements</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/VUPV_Parts"><strong>Go to Parts &rarr;</strong></a></div></br></br></br><br />
<br />
</br></br></div><br />
<div id="space-margin"></div><br />
<br />
</html><br />
<br />
{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Medal_requierementTeam:Valencia UPV/Medal requierement2014-10-18T03:38:15Z<p>Alquiru: </p>
<hr />
<div>{{:Team:Valencia_UPV/header}}<br />
<br />
<html><br />
<div align="center"><div id="cn-box" align="justify"><br />
<h3 class="hook" align="left"><a>Achievements</a> > <a>Medal Requirements</a></h3></br><br />
<br />
<div align="center"><span class="coda"><roja>M</roja>edal <roja>R</roja>equirements</span> </div><br/><br />
</br><br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/2/2b/VUPVEdiGEM_bronze_medal.png" alt="bronze_medal"></img><span class="background_medal"><b>Bronze Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="bronze"><br />
<li><a class="black-bold">Team registration</a></li><br/><br />
<li><a class="black-bold">Team Wiki</a></li><br/><br />
<li><a class="black-bold">Complete <a class="normal-link-page"href=" https://igem.org/2014_Judging_Form?id=1554">Judging Form</a> </a></li><br/><br />
<br />
<br />
<li><b>Present a poster and a talk at the iGEM Jamboree</b></li><br />
Poster ready to go!<br/><br/><br />
<li><b>The description of each project must clearly attribute work done by the students and distinguish it from work done by others, including host labs, advisors, instructors, sponsors, professional website designers, artists, and commercial services. </b></li><br />
<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Team/Attributions">See our Attributions here!</a><br/><br/><br />
<br />
<li><b> Document and submit new standard BioBrick Parts or Device to the iGEM Registry .<br/> <br />
<a class="normal-link-page"href="https://2014.igem.org/Team:Valencia_UPV/VUPV_Parts">See our Parts here!</a></b></li><br/> <br />
</ul><br />
<br/><br/><br />
<br />
<br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/5/5d/VUPVEdiGEM_silver_medal.png" alt="silver_medal"></img><span class="background_medal"><b>Silver Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="silver"><br />
<li><b>Experimentally validate that your new BioBrick Parts or Devices work as expected:</b><br/><br />
We can proudly assure that the following BioBricks work perfectly:<br />
<ul><br />
<li>BBa_K1554001, BBa_K1554002, BBa_K1554003 are the three enzymes that constitute the pheromone synthesis pathway. They proved to work and provided the expected results (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis">Results: Pheromone Analysis</a>).</li><br />
<li>BBa_K1554006 allows an efficient conversion from GoldenBraid 2.0 standard parts to BioBrick standard parts (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/parts_construction">Parts Contruction</a>).</li><br />
</ul></li><br/><br />
<li><b> Document the characterization of these parts in the “Main Page” section of that Part’s/Device’s Registry entry:</b><br/><br />
Check it! <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554001">BBa_K1554001</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554002">BBa_K1554002</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554003">BBa_K1554003</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554006">BBa_K1554006</a></li><br/><br />
<li><b>Submit this new part to the iGEM Parts <br/><br />
<a class="normal-link-page" href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2014&group=Valencia_UPV">Registry</a></b></li><br/><br />
<li><b>Articulate questions encountered by your team, and describe how you considered them within your project</b></li><br />
<li style="list-style: none;">To tune our project and ensure its outcome is in line with societal expectations, we engaged a wide range of social actors and stakeholders throughout the whole of it. Visit our <a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/policy/overview">Policy and Practices</a> section.</li><br/><br />
</ul><br />
<br/><br/><br />
<br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/8/87/VUPVEdiGEM_gold_medal.png" alt="gold_medal"></img><span class="background_medal"><b>Gold Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="gold"><br />
<li><b>Help any registered iGEM team from another school or institution by, for example, characterizing a part, debugging a construct, or modeling or simulating their system</b></li><br/><br />
<li style="list-style: none;">We developed a tight collaboration with the <a href="https://2014.igem.org/Team:NRP-UEA-Norwich" class="normal-link-page">NRP-UEA-Norwich</a> team. In collaboration, our team developed two biosafety devices thanks to two parts provided by them. Both devices were submitted to the iGEM Registry BBa_K1554004 and BBa_K1554005. In addition, the NRP-UEA-Norwich also provided us with their <a href="https://2014.igem.org/Team:NRP-UEA-Norwich/Project_Mo-Flipper" class="normal-link-page">Mo-Flippers</a>. We tested them in GoldenBraid standard parts and they worked perfectly.</li><br/><br />
</br></br></br><br/><br />
<br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/interlab"><strong>&larr; Go to Interlab Study</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Achievements"><strong>Go to Achievements</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/VUPV_Parts"><strong>Go to Parts &rarr;</strong></a></div></br></br></br><br />
<br />
</br></br></div><br />
<div id="space-margin"></div><br />
<br />
</html><br />
<br />
{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Medal_requierementTeam:Valencia UPV/Medal requierement2014-10-18T03:36:37Z<p>Alquiru: Undo revision 398001 by Alquiru (talk)</p>
<hr />
<div>{{:Team:Valencia_UPV/header}}<br />
<br />
<html><br />
<div align="center"><div id="cn-box" align="justify"><br />
<h3 class="hook" align="left"><a>Achievements</a> > <a>Medal Requirements</a></h3></br><br />
<br />
<div align="center"><span class="coda"><roja>M</roja>edal <roja>R</roja>equirements</span> </div><br/><br />
</br><br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/2/2b/VUPVEdiGEM_bronze_medal.png" alt="bronze_medal"></img><span class="background_medal"><b>Bronze Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="bronze"><br />
<li><a class="black-bold">Team registration</a></li><br/><br />
<li><a class="black-bold">Team Wiki</a></li><br/><br />
<li><a class="black-bold">Complete <a class="normal-link-page"href=" https://igem.org/2014_Judging_Form?id=1554">Judging Form</a> </a></li><br/><br />
<br />
<br />
<li><b>Present a poster and a talk at the iGEM Jamboree</b></li><br />
Poster ready to go!<br/><br/><br />
<li><b>The description of each project must clearly attribute work done by the students and distinguish it from work done by others, including host labs, advisors, instructors, sponsors, professional website designers, artists, and commercial services. </b></li><br />
<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Team/Attributions">See our Attributions here!</a><br/><br/><br />
<br />
<li><b> Document and submit new standard BioBrick Parts or Device to the iGEM Registry .<br/> <br />
<a class="normal-link-page"href="https://2014.igem.org/Team:Valencia_UPV/VUPV_Parts">See our Parts here!</a></b></li><br/> <br />
</ul><br />
<br/><br/><br />
<br />
<br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/5/5d/VUPVEdiGEM_silver_medal.png" alt="silver_medal"></img><span class="background_medal"><b>Silver Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="silver"><br />
<li><b>Experimentally validate that your new BioBrick Parts or Devices work as expected:</b><br/><br />
We can proudly assure that the following BioBricks work perfectly:<br />
<ul><br />
<li>BBa_K1554001, BBa_K1554002, BBa_K1554003 are the three enzymes that constitute the pheromone synthesis pathway. They proved to work and provided the expected results (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis">Results: Pheromone Analysis</a>).</li><br />
<li>BBa_K1554006 allows an efficient conversion from GoldenBraid 2.0 standard parts to BioBrick standard parts (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/parts_construction">Parts Contruction</a>).</li><br />
</ul></li><br/><br />
<li><b> Document the characterization of these parts in the “Main Page” section of that Part’s/Device’s Registry entry:</b><br/><br />
Check it! <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554001">BBa_K1554001</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554002">BBa_K1554002</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554003">BBa_K1554003</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554006">BBa_K1554006</a></li><br/><br />
<li><b>Submit this new part to the iGEM Parts <br/><br />
<a class="normal-link-page" href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2014&group=Valencia_UPV">Registry</a></b></li><br/><br />
<li><b>Articulate questions encountered by your team, and describe how you considered them within your project</b></li><br />
<li style="list-style: none;">In our <a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/policy/activities">Policy and Practices</a>, we extensively explore the pressing issues surrounding synthetic biology: 'Environmental Impact', 'Intellectual Property' 'Communication' and 'Public Engagement'.</li><br/><br />
</ul><br />
<br/><br/><br />
<br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/8/87/VUPVEdiGEM_gold_medal.png" alt="gold_medal"></img><span class="background_medal"><b>Gold Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="gold"><br />
<li><b>Help any registered iGEM team from another school or institution by, for example, characterizing a part, debugging a construct, or modeling or simulating their system</b></li><br/><br />
<li style="list-style: none;">We developed a tight collaboration with the <a href="https://2014.igem.org/Team:NRP-UEA-Norwich" class="normal-link-page">NRP-UEA-Norwich</a> team. In collaboration, our team developed two biosafety devices thanks to two parts provided by them. Both devices were submitted to the iGEM Registry BBa_K1554004 and BBa_K1554005. In addition, the NRP-UEA-Norwich also provided us with their <a href="https://2014.igem.org/Team:NRP-UEA-Norwich/Project_Mo-Flipper" class="normal-link-page">Mo-Flippers</a>. We tested them in GoldenBraid standard parts and they worked perfectly.</li><br/><br />
</br></br></br><br/><br />
<br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/interlab"><strong>&larr; Go to Interlab Study</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Achievements"><strong>Go to Achievements</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/VUPV_Parts"><strong>Go to Parts &rarr;</strong></a></div></br></br></br><br />
<br />
</br></br></div><br />
<div id="space-margin"></div><br />
<br />
</html><br />
<br />
{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Medal_requierementTeam:Valencia UPV/Medal requierement2014-10-18T03:32:56Z<p>Alquiru: </p>
<hr />
<div>{{:Team:Valencia_UPV/header}}<br />
<br />
<html><br />
<div align="center"><div id="cn-box" align="justify"><br />
<h3 class="hook" align="left"><a>Achievements</a> > <a>Medal Requirements</a></h3></br><br />
<br />
<div align="center"><span class="coda"><roja>M</roja>edal <roja>R</roja>equirements</span> </div><br/><br />
</br><br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/2/2b/VUPVEdiGEM_bronze_medal.png" alt="bronze_medal"></img><span class="background_medal"><b>Bronze Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="bronze"><br />
<li><a class="black-bold">Team registration</a></li><br/><br />
<li><a class="black-bold">Team Wiki</a></li><br/><br />
<li><a class="black-bold">Complete <a class="normal-link-page"href=" https://igem.org/2014_Judging_Form?id=1554">Judging Form</a> </a></li><br/><br />
<br />
<br />
<li><b>Present a poster and a talk at the iGEM Jamboree</b></li><br />
Poster ready to go!<br/><br/><br />
<li><b>The description of each project must clearly attribute work done by the students and distinguish it from work done by others, including host labs, advisors, instructors, sponsors, professional website designers, artists, and commercial services. </b></li><br />
<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Team/Attributions">See our Attributions here!</a><br/><br/><br />
<br />
<li><b> Document and submit new standard BioBrick Parts or Device to the iGEM Registry .<br/> <br />
<a class="normal-link-page"href="https://2014.igem.org/Team:Valencia_UPV/VUPV_Parts">See our Parts here!</a></b></li><br/> <br />
</ul><br />
<br/><br/><br />
<br />
<br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/5/5d/VUPVEdiGEM_silver_medal.png" alt="silver_medal"></img><span class="background_medal"><b>Silver Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="silver"><br />
<li><b>Experimentally validate that your new BioBrick Parts or Devices work as expected:</b><br/><br />
We can proudly assure that the following BioBricks work perfectly:<br />
<ul><br />
<li>BBa_K1554001, BBa_K1554002, BBa_K1554003 are the three enzymes that constitute the pheromone synthesis pathway. They proved to work and provided the expected results (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis">Results: Pheromone Analysis</a>).</li><br />
<li>BBa_K1554006 allows an efficient conversion from GoldenBraid 2.0 standard parts to BioBrick standard parts (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/parts_construction">Parts Contruction</a>).</li><br />
</ul></li><br/><br />
<li><b> Document the characterization of these parts in the “Main Page” section of that Part’s/Device’s Registry entry:</b><br/><br />
Check it! <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554001">BBa_K1554001</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554002">BBa_K1554002</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554003">BBa_K1554003</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554006">BBa_K1554006</a></li><br/><br />
<li><b>Submit this new part to the iGEM Parts <br/><br />
<a class="normal-link-page" href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2014&group=Valencia_UPV">Registry</a></b></li><br/><br />
<li><b>Articulate questions encountered by your team, and describe how you considered them within your project</b></li><br />
<li style="list-style: none;">In our <a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/policy/overview</a>, we extensively explore the pressing issues surrounding synthetic biology: 'Environmental Impact', 'Communication' and 'Public Engagement'.</li><br/><br />
</ul><br />
<br/><br/><br />
<br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/8/87/VUPVEdiGEM_gold_medal.png" alt="gold_medal"></img><span class="background_medal"><b>Gold Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="gold"><br />
<li><b>Help any registered iGEM team from another school or institution by, for example, characterizing a part, debugging a construct, or modeling or simulating their system</b></li><br/><br />
<li style="list-style: none;">We developed a tight collaboration with the <a href="https://2014.igem.org/Team:NRP-UEA-Norwich" class="normal-link-page">NRP-UEA-Norwich</a> team. In collaboration, our team developed two biosafety devices thanks to two parts provided by them. Both devices were submitted to the iGEM Registry BBa_K1554004 and BBa_K1554005. In addition, the NRP-UEA-Norwich also provided us with their <a href="https://2014.igem.org/Team:NRP-UEA-Norwich/Project_Mo-Flipper" class="normal-link-page">Mo-Flippers</a>. We tested them in GoldenBraid standard parts and they worked perfectly.</li><br/><br />
</br></br></br><br/><br />
<br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/interlab"><strong>&larr; Go to Interlab Study</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Achievements"><strong>Go to Achievements</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/VUPV_Parts"><strong>Go to Parts &rarr;</strong></a></div></br></br></br><br />
<br />
</br></br></div><br />
<div id="space-margin"></div><br />
<br />
</html><br />
<br />
{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Medal_requierementTeam:Valencia UPV/Medal requierement2014-10-18T03:29:52Z<p>Alquiru: </p>
<hr />
<div>{{:Team:Valencia_UPV/header}}<br />
<br />
<html><br />
<div align="center"><div id="cn-box" align="justify"><br />
<h3 class="hook" align="left"><a>Achievements</a> > <a>Medal Requirements</a></h3></br><br />
<br />
<div align="center"><span class="coda"><roja>M</roja>edal <roja>R</roja>equirements</span> </div><br/><br />
</br><br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/2/2b/VUPVEdiGEM_bronze_medal.png" alt="bronze_medal"></img><span class="background_medal"><b>Bronze Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="bronze"><br />
<li><a class="black-bold">Team registration</a></li><br/><br />
<li><a class="black-bold">Team Wiki</a></li><br/><br />
<li><a class="black-bold">Complete <a class="normal-link-page"href=" https://igem.org/2014_Judging_Form?id=1554">Judging Form</a> </a></li><br/><br />
<br />
<br />
<li><b>Present a poster and a talk at the iGEM Jamboree</b></li><br />
Poster ready to go!<br/><br/><br />
<li><b>The description of each project must clearly attribute work done by the students and distinguish it from work done by others, including host labs, advisors, instructors, sponsors, professional website designers, artists, and commercial services. </b></li><br />
<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Team/Attributions">See our Attributions here!</a><br/><br/><br />
<br />
<li><b> Document and submit new standard BioBrick Parts or Device to the iGEM Registry .<br/> <br />
<a class="normal-link-page"href="https://2014.igem.org/Team:Valencia_UPV/VUPV_Parts">See our Parts here!</a></b></li><br/> <br />
</ul><br />
<br/><br/><br />
<br />
<br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/5/5d/VUPVEdiGEM_silver_medal.png" alt="silver_medal"></img><span class="background_medal"><b>Silver Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="silver"><br />
<li><b>Experimentally validate that your new BioBrick Parts or Devices work as expected:</b><br/><br />
We can proudly assure that the following BioBricks work perfectly:<br />
<ul><br />
<li>BBa_K1554001, BBa_K1554002, BBa_K1554003 are the three enzymes that constitute the pheromone synthesis pathway. They proved to work and provided the expected results (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis">Results: Pheromone Analysis</a>).</li><br />
<li>BBa_K1554006 allows an efficient conversion from GoldenBraid 2.0 standard parts to BioBrick standard parts (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/parts_construction">Parts Contruction</a>).</li><br />
</ul></li><br/><br />
<li><b> Document the characterization of these parts in the “Main Page” section of that Part’s/Device’s Registry entry:</b><br/><br />
Check it! <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554001">BBa_K1554001</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554002">BBa_K1554002</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554003">BBa_K1554003</a>, <a class="normal-link-page" href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554006">BBa_K1554006</a></li><br/><br />
<li><b>Submit this new part to the iGEM Parts <br/><br />
<a class="normal-link-page" href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2014&group=Valencia_UPV">Registry</a></b></li><br/><br />
<li><b>Articulate questions encountered by your team, and describe how you considered them within your project</b></li><br />
<li style="list-style: none;">In our <a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/policy/activities">Policy and Practices</a>, we extensively explore the pressing issues surrounding synthetic biology: 'Environmental Impact', 'Intellectual Property' 'Communication' and 'Public Engagement'.</li><br/><br />
</ul><br />
<br/><br/><br />
<br />
<br />
<img style="z-index: 15;" width="120px" src="https://static.igem.org/mediawiki/2014/8/87/VUPVEdiGEM_gold_medal.png" alt="gold_medal"></img><span class="background_medal"><b>Gold Medal Requirements</b></span><br/><br />
<br />
<ul class="medals-tick" id="gold"><br />
<li><b>Help any registered iGEM team from another school or institution by, for example, characterizing a part, debugging a construct, or modeling or simulating their system</b></li><br/><br />
<li style="list-style: none;">We developed a tight collaboration with the <a href="https://2014.igem.org/Team:NRP-UEA-Norwich" class="normal-link-page">NRP-UEA-Norwich</a> team. In collaboration, our team developed two biosafety devices thanks to two parts provided by them. Both devices were submitted to the iGEM Registry BBa_K1554004 and BBa_K1554005. In addition, the NRP-UEA-Norwich also provided us with their <a href="https://2014.igem.org/Team:NRP-UEA-Norwich/Project_Mo-Flipper" class="normal-link-page">Mo-Flippers</a>. We tested them in GoldenBraid standard parts and they worked perfectly.</li><br/><br />
</br></br></br><br/><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafetyTeam:Valencia UPV/Project/modules/biosafety2014-10-18T03:27:40Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a>Biosafety</a> </h3></p><br/><br />
<br />
<div align="center"><span class="coda"><roja>B</roja>iosafety</span> </div><br/><br/><br />
<br />
<p>"With great powers comes great responsibility." – <span class="italic">Benjamin Parker (Uncle Ben)</span>.</p><br/><br/><br />
<br />
<p>We know the importance of keeping genetically modified organisms under control. Ideally, modified genetic material should be unable to spread and organisms containing it should be easily distinguishable from wild type. Keeping this in mind, we created a biosafety module to be used in plants which will be available for future iGEM teams working with plants. Our biosafety module combines two biosafety strategies: <span class="black-bold">identity preservation</span> and <span class="black-bold">male sterility</span>.</p><br/><br/><br />
<br />
<br />
<br />
<br />
<p><span class="black-bold">Identity preservation</span> enables an easy identification of the genetically modified organism. We expressed SlANT1 and SlJAF13 genes from tomato, which code for transcription factors that activate synthesis of <span class="black-bold">anthocyanines </span> [1], coloured compounds found in plants. The introduction of this element in the biosafety module generates differentially coloured plants (Figures 1 and 2). <a class="normal-link-page" target="_blank" href="https://2014.igem.org/Team:NRP-UEA-Norwich">NRP-UEA-Norwich</a> , and it is also used in their project “<span class="italic">Green Canary</span>”.</p><br/><br/><br />
<br />
<br />
<br />
<div align="center"><img height= "170px;" style="margin-right: 30px;" src="https://static.igem.org/mediawiki/2014/b/b8/VUPV_Rosea.png" alt="N benthamiana Rosea" title=N benthamiana Rosea""></img><img width="250px" style="margin-left: 30px;" src="https://static.igem.org/mediawiki/2014/5/57/VUPVhoja.png" alt="Dark leaf" title="Dark leaf"></img><br/><br/><br />
<p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>.Anthocyanin accumulation in a <i>N benthamiana</i> plant. <br />
<p style="text-align: right; font-style: italic; font-size: 0.8em; width: 700px;"><b>Figure 2</b>. Dark purple leaf with induced anthocyanin accumulation. </p><br/><br />
<br />
</div><br />
<p><span class="black-bold">Male sterility</span> makes impossible the dispersion of genetic material using pollen as the vehicle or by seeds result of self-pollination. In order to achieve this dispersion restriction, we integrated the active peptide of <span class="black-bold">barnase</span>, a RNAse from <span class="italic">Bacillus amyloliquefaciens</span>, (Biobricks accession code BBa_I716211) under the regulation of the tapetum-specific promoter TA29 [1]. Both components are very well documented since TA29 has been used by a large number of researchers [2-6] and barnase has also been used under the regulation of different promoters [7,8]. We chose this strategy because it had been previously used in our laboratory with satisfactory results [9].</p><br/><br />
<br />
<p>Our Biosafety module is our major present for future iGEM teams. It can be applied to a broad variety of new systems in future projects. We are certain that future iGEMers will realize how important biosafety is and they will incorporate our biosafety module in their systems.</p><br />
<br/><br />
<br />
<br />
</ol></br></br><br />
<br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><strong>&larr; Go to Methodology</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules"><strong>Go to Modules</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/ideal_plant"><strong>Go to Future Perspectives &rarr;</strong></a></div></br></br></br><br />
<br />
<br />
<br/><br/><p align="center"><strong>References</strong></p><br/><br />
<div style="position: relative; left: 3%; width: 96%;"><ol><br />
<li>Mariani C, Beuckeleer MD, Truettner J, Leemans J, Goldberg RB (1990) Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347: 737-741.</li><br />
<li>Cho HJ, Kim S, Kim M, Kim BD (2001) Production of transgenic male sterile tobacco plants with the cDNA encoding a ribosome inactivating protein in Dianthus sinensis L. Mol Cells 11: 326-333.</li><br />
<li>Sa G, Mi M, He-Chun Y, Guo-Feng L (2002) Anther-specific expression of ipt gene in transgenic tobacco and its effect on plant development. Transgenic Res 11: 269-278.</li><br />
<li>Shaya F, Gaiduk S, Keren I, Shevtsov S, Zemah H, et al. (2012) Expression of mitochondrial gene fragments within the tapetum induce male sterility by limiting the biogenesis of the respiratory machinery in transgenic tobacco. J Integr Plant Biol 54: 115-130.</li><br />
<li>Kriete G, Niehaus K, Perlick AM, Puhler A, Broer I (1996) Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compound N-acetyl-L-phosphinothricin. Plant J 9: 809-818.</li><br />
<li>Shukla P, Singh NK, Kumar D, Vijayan S, Ahmed I, et al. (2014) Expression of a pathogen-induced cysteine protease (AdCP) in tapetum results in male sterility in transgenic tobacco. Funct Integr Genomics 14: 307-317.</li><br />
<li>Goldman MH, Goldberg RB, Mariani C (1994) Female sterile tobacco plants are produced by stigma-specific cell ablation. EMBO J 13: 2976-2984.</li><br />
<br />
<li>Wang HZ, Hu B, Chen GP, Shi NN, Zhao Y, et al. (2008) Application of Arabidopsis AGAMOUS second intron for the engineered ablation of flower development in transgenic tobacco. Plant Cell Rep 27: 251-259.</li><br />
<br />
<li>Sarrion-Perdigones A, Falconi EE, Zandalinas SI, Juarez P, Fernandez-del-Carmen A, et al. (2011) GoldenBraid: an iterative cloning system for standardized assembly of reusable genetic modules. PLoS One 6: e21622.</li><br />
</div><br />
</br></br></div><br />
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</html><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafetyTeam:Valencia UPV/Project/modules/biosafety2014-10-18T03:25:01Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a>Biosafety</a> </h3></p><br/><br />
<br />
<div align="center"><span class="coda"><roja>B</roja>iosafety</span> </div><br/><br/><br />
<br />
<p>"With great powers comes great responsibility." – <span class="italic">Benjamin Parker (Uncle Ben)</span>.</p><br/><br/><br />
<br />
<p>We know the importance of keeping genetically modified organisms under control. Ideally, modified genetic material should be unable to spread and organisms containing it should be easily distinguishable from wild type. Keeping this in mind, we created a biosafety module to be used in plants which will be available for future iGEM teams working with plants. Our biosafety module combines two biosafety strategies: <span class="black-bold">identity preservation</span> and <span class="black-bold">male sterility</span>.</p><br/><br/><br />
<br />
<br />
<br />
<br />
<p><span class="black-bold">Identity preservation</span> enables an easy identification of the genetically modified organism. We expressed SlANT1 and SlJAF13 genes from tomato, which code for transcription factors that activate synthesis of <span class="black-bold">anthocyanines </span> [1], coloured compounds found in plants. The introduction of this element in the biosafety module generates differentially coloured plants (Figures 1 and 2). <a class="normal-link-page" target="_blank" href="https://2014.igem.org/Team:NRP-UEA-Norwich">NRP-UEA-Norwich</a> , and it is also used in their project “<span class="italic">Green Canary</span>”.</p><br/><br/><br />
<br />
<br />
<br />
<div align="center"><img height= "170px;" style="margin-right: 30px;" src="https://static.igem.org/mediawiki/2014/b/b8/VUPV_Rosea.png" alt="N benthamiana Rosea" title=N benthamiana Rosea""></img><img width="250px" style="margin-left: 30px;" src="https://static.igem.org/mediawiki/2014/5/57/VUPVhoja.png" alt="Dark leaf" title="Dark leaf"></img><br/><br/><br />
<p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>.Anthocyanin accumulation in a <i>N benthamiana</i> plant. <br />
<p style="text-align: right; font-style: italic; font-size: 0.8em; width: 700px;"><b>Figure 2</b>. Dark purple leaf with induced anthocyanin accumulation. </p><br/><br />
<br />
</div><br />
<p><span class="black-bold">Male sterility</span> makes impossible the dispersion of genetic material using pollen as the vehicle or by seeds result of self-pollination. In order to achieve this dispersion restriction, we integrated the active peptide of <span class="black-bold">barnase</span>, a RNAse from <span class="italic">Bacillus amyloliquefaciens</span>, (Biobricks accession code BBa_I716211) under the regulation of the tapetum-specific promoter TA29 [1]. Both components are very well documented since TA29 has been used by a large number of researchers [2-6] and barnase has also been used under the regulation of different promoters [7,8]. We chose this strategy because it had been previously used in our laboratory with satisfactory results [9].</p><br/><br />
<br />
<p>Our Biosafety module is our major present for future iGEM teams. It can be applied to a broad variety of new systems in future projects. We are certain that future iGEMers will realize how important biosafety is and they will incorporate our biosafety module in their systems.</p><br />
<br/><br />
<br />
<p align="center"><strong>References</strong></p><br/><br />
<div style="position: relative; left: 3%; width: 96%;"><ol><br />
<li>Mariani C, Beuckeleer MD, Truettner J, Leemans J, Goldberg RB (1990) Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347: 737-741.</li><br />
<li>Cho HJ, Kim S, Kim M, Kim BD (2001) Production of transgenic male sterile tobacco plants with the cDNA encoding a ribosome inactivating protein in Dianthus sinensis L. Mol Cells 11: 326-333.</li><br />
<li>Sa G, Mi M, He-Chun Y, Guo-Feng L (2002) Anther-specific expression of ipt gene in transgenic tobacco and its effect on plant development. Transgenic Res 11: 269-278.</li><br />
<li>Shaya F, Gaiduk S, Keren I, Shevtsov S, Zemah H, et al. (2012) Expression of mitochondrial gene fragments within the tapetum induce male sterility by limiting the biogenesis of the respiratory machinery in transgenic tobacco. J Integr Plant Biol 54: 115-130.</li><br />
<li>Kriete G, Niehaus K, Perlick AM, Puhler A, Broer I (1996) Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compound N-acetyl-L-phosphinothricin. Plant J 9: 809-818.</li><br />
<li>Shukla P, Singh NK, Kumar D, Vijayan S, Ahmed I, et al. (2014) Expression of a pathogen-induced cysteine protease (AdCP) in tapetum results in male sterility in transgenic tobacco. Funct Integr Genomics 14: 307-317.</li><br />
<li>Goldman MH, Goldberg RB, Mariani C (1994) Female sterile tobacco plants are produced by stigma-specific cell ablation. EMBO J 13: 2976-2984.</li><br />
<br />
<li>Wang HZ, Hu B, Chen GP, Shi NN, Zhao Y, et al. (2008) Application of Arabidopsis AGAMOUS second intron for the engineered ablation of flower development in transgenic tobacco. Plant Cell Rep 27: 251-259.</li><br />
<br />
<li>Sarrion-Perdigones A, Falconi EE, Zandalinas SI, Juarez P, Fernandez-del-Carmen A, et al. (2011) GoldenBraid: an iterative cloning system for standardized assembly of reusable genetic modules. PLoS One 6: e21622.</li> <br />
</ol></br></br><br/><br/><br />
<br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><strong>&larr; Go to Methodology</strong></a><br />
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<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/ideal_plant"><strong>Go to Future Perspectives &rarr;</strong></a></div></br></br></br><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafetyTeam:Valencia UPV/Project/modules/biosafety2014-10-18T03:17:17Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a>Biosafety</a> </h3></p><br/><br />
<br />
<div align="center"><span class="coda"><roja>B</roja>iosafety</span> </div><br/><br/><br />
<br />
<p>"With great powers comes great responsibility." – <span class="italic">Benjamin Parker (Uncle Ben)</span>.</p><br/><br/><br />
<br />
<p>We know the importance of keeping genetically modified organisms under control. Ideally, modified genetic material should be unable to spread and organisms containing it should be easily distinguishable from wild type. Keeping this in mind, we created a biosafety module to be used in plants which will be available for future iGEM teams working with plants. Our biosafety module combines two biosafety strategies: <span class="black-bold">identity preservation</span> and <span class="black-bold">male sterility</span>.</p><br/><br/><br />
<br />
<br />
<br />
<br />
<p><span class="black-bold">Identity preservation</span> enables an easy identification of the genetically modified organism. We expressed SlANT1 and SlJAF13 genes from tomato, which code for transcription factors that activate synthesis of <span class="black-bold">anthocyanines </span> [1], coloured compounds found in plants. The introduction of this element in the biosafety module generates differentially coloured plants (Figures 1 and 2). <a class="normal-link-page" target="_blank" href="https://2014.igem.org/Team:NRP-UEA-Norwich">NRP-UEA-Norwich</a> , and it is also used in their project “<span class="italic">Green Canary</span>”.</p><br/><br/><br />
<br />
<br />
<br />
<div align="center"><img height= "170px;" style="margin-right: 30px;" src="https://static.igem.org/mediawiki/2014/b/b8/VUPV_Rosea.png" alt="N benthamiana Rosea" title=N benthamiana Rosea""></img><img width="250px" style="margin-left: 30px;" src="https://static.igem.org/mediawiki/2014/5/57/VUPVhoja.png" alt="Dark leaf" title="Dark leaf"></img><br/><br />
<p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>.Anthocyanin accumulation in a <i>N benthamiana</i> plant <p style="text-align: right; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 2</span>. Leaf anthocyanins accumulation </p><br />
<br />
</div><br />
<p><span class="black-bold">Male sterility</span> makes impossible the dispersion of genetic material using pollen as the vehicle or by seeds result of self-pollination. In order to achieve this dispersion restriction, we integrated the active peptide of <span class="black-bold">barnase</span>, a RNAse from <span class="italic">Bacillus amyloliquefaciens</span>, (Biobricks accession code BBa_I716211) under the regulation of the tapetum-specific promoter TA29 [1]. Both components are very well documented since TA29 has been used by a large number of researchers [2-6] and barnase has also been used under the regulation of different promoters [7,8]. We chose this strategy because it had been previously used in our laboratory with satisfactory results [9].</p><br/><br />
<br />
<p>Our Biosafety module is our major present for future iGEM teams. It can be applied to a broad variety of new systems in future projects. We are certain that future iGEMers will realize how important biosafety is and they will incorporate our biosafety module in their systems.</p><br />
<br/><br />
<br />
<p align="center"><strong>References</strong></p><br/><br />
<div style="position: relative; left: 3%; width: 96%;"><ol><br />
<li>Mariani C, Beuckeleer MD, Truettner J, Leemans J, Goldberg RB (1990) Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347: 737-741.</li><br />
<li>Cho HJ, Kim S, Kim M, Kim BD (2001) Production of transgenic male sterile tobacco plants with the cDNA encoding a ribosome inactivating protein in Dianthus sinensis L. Mol Cells 11: 326-333.</li><br />
<li>Sa G, Mi M, He-Chun Y, Guo-Feng L (2002) Anther-specific expression of ipt gene in transgenic tobacco and its effect on plant development. Transgenic Res 11: 269-278.</li><br />
<li>Shaya F, Gaiduk S, Keren I, Shevtsov S, Zemah H, et al. (2012) Expression of mitochondrial gene fragments within the tapetum induce male sterility by limiting the biogenesis of the respiratory machinery in transgenic tobacco. J Integr Plant Biol 54: 115-130.</li><br />
<li>Kriete G, Niehaus K, Perlick AM, Puhler A, Broer I (1996) Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compound N-acetyl-L-phosphinothricin. Plant J 9: 809-818.</li><br />
<li>Shukla P, Singh NK, Kumar D, Vijayan S, Ahmed I, et al. (2014) Expression of a pathogen-induced cysteine protease (AdCP) in tapetum results in male sterility in transgenic tobacco. Funct Integr Genomics 14: 307-317.</li><br />
<li>Goldman MH, Goldberg RB, Mariani C (1994) Female sterile tobacco plants are produced by stigma-specific cell ablation. EMBO J 13: 2976-2984.</li><br />
<br />
<li>Wang HZ, Hu B, Chen GP, Shi NN, Zhao Y, et al. (2008) Application of Arabidopsis AGAMOUS second intron for the engineered ablation of flower development in transgenic tobacco. Plant Cell Rep 27: 251-259.</li><br />
<br />
<li>Sarrion-Perdigones A, Falconi EE, Zandalinas SI, Juarez P, Fernandez-del-Carmen A, et al. (2011) GoldenBraid: an iterative cloning system for standardized assembly of reusable genetic modules. PLoS One 6: e21622.</li> <br />
</ol></br></br><br/><br/><br />
<br />
<div align="center"><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/File:VUPVhoja.pngFile:VUPVhoja.png2014-10-18T02:57:09Z<p>Alquiru: uploaded a new version of &quot;File:VUPVhoja.png&quot;</p>
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<div></div>Alquiruhttp://2014.igem.org/File:VUPVhoja.pngFile:VUPVhoja.png2014-10-18T02:55:13Z<p>Alquiru: </p>
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<div></div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modules/moths_behaviorTeam:Valencia UPV/Project/modules/moths behavior2014-10-18T02:51:41Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a>Moths Behaviour</a> </h3></p><br/><br/><br />
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<div align="center"><span class="coda"><roja>M</roja>oths <roja>B</roja>ehaviour<roja></span> </div><br/><br />
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<p class="subpart">The Idea</p><br/><br />
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<p>You better know your enemy before starting a fight. Moths, flying insects from the order Lepidoptera, cause great damage in crops all over the world. We aim to avoid damages caused by them, so understanding their behaviour is a main issue for us. We focused in their sexual behaviour to look for an elegant strategy to fight them.</p><br/><br/><br />
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<p class="subpart">Moth Sexual Behavior</p><br/><br />
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<p>Moth individuals within the same specie communicate with each other using volatile chemical compounds called “sex pheromones”. Female moths generate several pheromone compounds in their sex pheromone gland, at the tip of their abdomen, and release them in certain ratios. Male moths locate the females of their own species, by tracing these compounds in species-specific ratios [1]. Offspring derived from this encounter is the cause of damage in crops.</p><br/><br/><br />
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<p class="subpart">Pheromone-based Pest Control in Moths</p><br/><br />
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<div><br />
<img width="300px" style="float:right; margin-left: 15px;" src="https://static.igem.org/mediawiki/2014/f/fa/VUPVMoth.png" alt="moth_img"></img><br />
<p><br/>Some experts indicate that "The direct management of insect pests using pheromones can provide excellent suppression of key lepidopteran pests in agriculture" [2]. If a high concentration of the major pheromone component (the most abundant pheromone for a specie) is present in the air, this may lead to a "nonresponsive outcome". Fatigue or adaptation of the male's pheromone sensory organs are the cause of this [2]. This strategy is called <b>mating disruption</b>.</p><br/><br/><br />
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<br/><p style="text-align: right; font-style: italic; font-size: 0.8em; width: 775px;"><span class="black-bold">Figure 1</span>. <i>Helicoverpa armigers</i></p><br />
<p style="text-align: right; font-style: italic; font-size: 0.5em; width: 775px;"><span class="black-bold">Source:</span>. <i>http://commons.wikimedia.org/wiki/File:Noctuidae_-_Helicoverpa_armigera.JPG</i></p></div><br />
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<p>Traditional pest control strategies based on pesticides are non-specific and affect the target along with other species (bees, humans, etc.), which can cause ecological problems [3]. However, <b>pheromone-based pest control strategies</b> are more species-specific than pesticides and they target a narrower range of species. They have shown no effect on pest species outside of a "cropping system, reducing the risk of outbreak of a secondary pest.</p><br/><br/><br/><br />
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<p><span class="red-bold">Sexy Plant</span> uses this pheromone-based mating-disruption strategy to avoid damages in crops and becomes an environmentally-friendly choice in pest control.</p><br/><br/><br/><br/><br />
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<p align="center"><strong>References</strong></p><br/><br />
<div style="position: relative; left: 3%; width: 96%;"><ol><br />
<li>Vogel H, Heidel AJ, Heckel DG, Groot AT (2010) Transcriptome analysis of the sex pheromone gland of the noctuid moth Heliothis virescens. BMC Genomics 11: 29.</li><br />
<li>Welter SC, Pickel C, Millar J, Cave F, Steenwyk RAV, et al. (2005) Pheromone mating disruption offers selective management options for key pests. California Agriculture 59: 16-22.</li><br />
<li>Brittain C, Potts SG (2011) The potential impacts of insecticides on the life-history traits of bees and the consequences for pollination. Basic and Applied Ecology 12: 321-331.</li><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafetyTeam:Valencia UPV/Project/modules/biosafety2014-10-18T02:45:55Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a>Biosafety</a> </h3></p><br/><br />
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<div align="center"><span class="coda"><roja>B</roja>iosafety</span> </div><br/><br/><br />
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<p>"With great powers comes great responsibility." – <span class="italic">Benjamin Parker (Uncle Ben)</span>.</p><br/><br/><br />
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<p>We know the importance of keeping genetically modified organisms under control. Ideally, modified genetic material should be unable to spread and organisms containing it should be easily distinguishable from wild type. Keeping this in mind, we created a biosafety module to be used in plants which will be available for future iGEM teams working with plants. Our biosafety module combines two biosafety strategies: <span class="black-bold">identity preservation</span> and <span class="black-bold">male sterility</span>.</p><br/><br/><br />
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<p><span class="black-bold">Identity preservation</span> enables an easy identification of the genetically modified organism. We expressed SlANT1 and SlJAF13 genes from tomato, which code for transcription factors that activate synthesis of <span class="black-bold">anthocyanines </span> [1], coloured compounds found in plants (Figure 1). The introduction of this element in the biosafety module generates differentially coloured plants (Figures 2 and 3). <a class="normal-link-page" target="_blank" href="https://2014.igem.org/Team:NRP-UEA-Norwich">NRP-UEA-Norwich</a> , and it is also used in their project “<span class="italic">Green Canary</span>”.</p><br/><br/><br />
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<p><span class="black-bold">Male sterility</span> makes impossible the dispersion of genetic material using pollen as the vehicle or by seeds result of self-pollination. In order to achieve this dispersion restriction, we integrated the active peptide of <span class="black-bold">barnase</span>, a RNAse from <span class="italic">Bacillus amyloliquefaciens</span>, (Biobricks accession code BBa_I716211) under the regulation of the tapetum-specific promoter TA29 [1]. Both components are very well documented since TA29 has been used by a large number of researchers [2-6] and barnase has also been used under the regulation of different promoters [7,8]. We chose this strategy because it had been previously used in our laboratory with satisfactory results [9].</p><br/><br />
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<p>Our Biosafety module is our major present for future iGEM teams. It can be applied to a broad variety of new systems in future projects. We are certain that future iGEMers will realize how important biosafety is and they will incorporate our biosafety module in their systems.</p><br />
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<p align="center"><strong>References</strong></p><br/><br />
<div style="position: relative; left: 3%; width: 96%;"><ol><br />
<li>Mariani C, Beuckeleer MD, Truettner J, Leemans J, Goldberg RB (1990) Induction of male sterility in plants by a chimaeric ribonuclease gene. Nature 347: 737-741.</li><br />
<li>Cho HJ, Kim S, Kim M, Kim BD (2001) Production of transgenic male sterile tobacco plants with the cDNA encoding a ribosome inactivating protein in Dianthus sinensis L. Mol Cells 11: 326-333.</li><br />
<li>Sa G, Mi M, He-Chun Y, Guo-Feng L (2002) Anther-specific expression of ipt gene in transgenic tobacco and its effect on plant development. Transgenic Res 11: 269-278.</li><br />
<li>Shaya F, Gaiduk S, Keren I, Shevtsov S, Zemah H, et al. (2012) Expression of mitochondrial gene fragments within the tapetum induce male sterility by limiting the biogenesis of the respiratory machinery in transgenic tobacco. J Integr Plant Biol 54: 115-130.</li><br />
<li>Kriete G, Niehaus K, Perlick AM, Puhler A, Broer I (1996) Male sterility in transgenic tobacco plants induced by tapetum-specific deacetylation of the externally applied non-toxic compound N-acetyl-L-phosphinothricin. Plant J 9: 809-818.</li><br />
<li>Shukla P, Singh NK, Kumar D, Vijayan S, Ahmed I, et al. (2014) Expression of a pathogen-induced cysteine protease (AdCP) in tapetum results in male sterility in transgenic tobacco. Funct Integr Genomics 14: 307-317.</li><br />
<li>Goldman MH, Goldberg RB, Mariani C (1994) Female sterile tobacco plants are produced by stigma-specific cell ablation. EMBO J 13: 2976-2984.</li><br />
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<li>Wang HZ, Hu B, Chen GP, Shi NN, Zhao Y, et al. (2008) Application of Arabidopsis AGAMOUS second intron for the engineered ablation of flower development in transgenic tobacco. Plant Cell Rep 27: 251-259.</li><br />
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<li>Sarrion-Perdigones A, Falconi EE, Zandalinas SI, Juarez P, Fernandez-del-Carmen A, et al. (2011) GoldenBraid: an iterative cloning system for standardized assembly of reusable genetic modules. PLoS One 6: e21622.</li> <br />
</ol></br></br><br/><br/><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/File:VUPV_Rosea.pngFile:VUPV Rosea.png2014-10-18T02:43:15Z<p>Alquiru: uploaded a new version of &quot;File:VUPV Rosea.png&quot;</p>
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<div></div>Alquiruhttp://2014.igem.org/File:VUPVMoth.pngFile:VUPVMoth.png2014-10-18T02:19:50Z<p>Alquiru: </p>
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<div></div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/overviewTeam:Valencia UPV/Project/overview2014-10-18T02:15:09Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a>Overview</a></h3></p><br/><br />
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<div align="center"><span class="coda"><roja>P</roja>roject <roja>O</roja>verview</span> </div><br/><br/><br />
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<p>Pests cause <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/problem" class="normal-link-page">great economical loss</a> in agriculture and impede an optimal use of the resources. <span class="red-bold">Sexy Plant</span> rises as a pest control strategy based on the use of <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/moths_behavior" class="normal-link-page">mating disruption</a> by sex pheromones. The release of sex pheromones impedes male moths to find their females, avoiding crop-damaging larvae to be born.</p><br/><br />
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<p>The <span class="red-bold">Sexy Plant</span> is engineered to <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesis" class="normal-link-page">produce</a> and <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/release" class="normal-link-page">release</a> three moth sex pheromones which affect a broad number of Lepidoptera species (moths). We implemented previous work by Ding et al. [1] to produce the first two pheromones (<span class="red-bold">Z11-16:OH</span> and <span class="blue-bold">Z11-16:OAc</span>). Inspired by Hagström et al [2], we planned the introduction of a Fatty Acid reductase to transform <span class="red-bold">Z11-16:OH</span> to <span class="green-bold">Z11-16:Ald</span>. As result, the sexy plant will be able to produce three sex pheromones involved in moth’s mating disruption (Figure 1).</p><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/4/43/VUPVOv_fig2.png" alt="pheromone_pathway" title="Pheromone Pathway" width="450px"></img></div><br/><br />
<div align="center"><p style="text-align: center; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1. Biosynthetic pathway of moth sex pheromones</span>. Sex pheromones are bordered in purple.</p></div><br/><br />
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<p>Biosafety is a major concern in our project. For that reason, we have developed a <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety" class="normal-link-page">biosafety module</a> which allows an easy identification of the plant by identity preservation and prevents the plant to spread its genetic material via pollen.</p><br/><br />
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<p>In addition, we decided to enable external control of pheromone production, designing a <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/switch" class="normal-link-page">genetic switch</a> that controls the activation of the biosynthetic pathway. This genetic switch turns on the production of pheromones when a solution containing CuSO4 is sprayed on the plant.</p><br/><br />
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<p>The different genetic modules are put together to create the complete system. The system resembles an electric circuit, as it is represented in figure 2. Our genetic circuit, once introduced in <i>Nicotiana benthamiana</i> creates the <span class="red-bold">Sexy Plant</span>, a plant to fight moths and avoid damages in crops.</p><br/><br />
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<div align="center"><img width="400px" src="https://static.igem.org/mediawiki/2014/a/ac/VUPVOverview_figure_2.png" alt="circuit"></img></div><br/><br />
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<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 2. Our system in an electric circuit-like format</span>. The plant’s own metabolism is used as the energy source. The biosafety module, composed by identity preservation and male sterility parts is always under expression in the system, to ensure the security of the transgenic plant. Pheromone synthetic pathways (A,B or C, depending on the problem pest) are activated using a genetic switch sensitive to Copper Sulfate, so system will not be producing pheromones unless it is activated with Copper-rich solution.</p></div><br/><br/><br/><br/><br/><br/><br />
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<p align="center" class="black-bold">REFERENCES</p><br/><br />
<div style="position: relative; left: 3%; width: 96%;"><ol><br />
<li>Ding BJ, Hofvander P, Wang HL, Durrett TP, Stymne S, et al. (2014) A plant factory for moth pheromone production. Nat Commun 5: 3353.</li><br />
<li>Hagström A, Wang HL, Lienard MA, Lassance JM, Johansson T, et al. (2013) A moth pheromone brewery: production of (Z)-11-hexadecenol by heterologous co-expression of two biosynthetic genes from a noctuid moth in a yeast cell factory. Microb Cell Fact 12: 125.</li><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/resultsTeam:Valencia UPV/Project/results2014-10-18T02:09:53Z<p>Alquiru: </p>
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/safetyTeam:Valencia UPV/safety2014-10-18T02:02:41Z<p>Alquiru: </p>
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One of today’s greatest concerns regarding Genetically Modified Organisms (GMO) is biosafety. While talking to people about our project we realized that fears about GMO are still present in our society. Therefore, we paid special attention to public perception and understanding when discussing our project approach as explained in the <a href="https://2014.igem.org/Team:Valencia_UPV/policy/overview" class="normal-link-page">Policy and Practices overview.</a> <br/><br/><br />
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As a result, the <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety" class="normal-link-page">Biosafety module</a> was developed. This module has two objectives, to preserve the identity of the genetically engineered plant allowing its unequivocal identification by changing its color; and avoid its uncontrolled spread inhibiting the pollen production by male-sterility. We implemented this module in collaboration with NRP-UEA-Norwich team and <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/biosafety" class="normal-link-page">Results</a> can be checked here. <br/><br/><br />
Additionally, our team successfully completed the <a href="https://igem.org/Safety/Safety_Form?team_id=1554" class="normal-link-page">Safety form</a> updating it when it was necessary.<br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/AchievementsTeam:Valencia UPV/Achievements2014-10-18T01:37:47Z<p>Alquiru: </p>
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<div align="center"><span class="coda"><roja>A</roja>chievements</span> </div><br/><br/><br />
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<p>As it can bee observed surfing our wiki, at the end of this long road we have accomplished many positive results:</p><br />
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<ul class="method"><br />
<li><a class="black-bold">A plant able to produce three insect sexual pheromones from moths to produce insects mating disruption, the Sexy Plant.:</a> <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis" class="normal-link-page">Results: Pheromone analysis</a><br />
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<li>Obtained pheromones being among the most abundant plant organic volatile compounds.<span class="red-bold">(Z)-11-hexadecen-1-ol</span> is certainly the most abundant one.</li><br />
<li>Successful and functional assembly of each of the pheromone biosynthetic genes with the plant constitutive promoter P35S. Also multigenic assembly of all three transcription units in a single plasmid. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/contructs" class="normal-link-page">Results: Constructions-Biosynthesis</a></li><br />
<li>Proof of our produced pheromones-insect interaction. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/contructs" class="normal-link-page">Results: Electroantennography</a></li><br />
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<ul class="method"><br />
<li><a class="black-bold">A broad understanding of plant metabolism, by adapting and exploitation of a genome-scale model of Arabidopsis thaliana primary metabolism able to help us theoretically optimize the pheromone production </a> <a href="https://2014.igem.org/Team:Valencia_UPV/Modeling/fba" class="normal-link-page"> Modeling: Pheromone Production</a><br />
<ul><br />
<li>Adapting the AraGEM genome-scale model to our pheromone production pathway by branching the flux of our precursor metabolite Palmitic acic (16:0).</li><br />
<li>Identifying the principal: i) cell compartments and scenarios of the plant metabolism, ii) flux bounds and constraints for each scenario, and iii) the interactions between chemical reactions and substrates related to our pheromone. </li><br />
<li>Exploring genetic conditions that could improve our pheromone production by Single gene Knock-out analysis. </li><br />
<li>Obtaining optimal condition for the coupled yield of pheromone and biomass production as a function of photons consumption.</li> <br />
</ul><br />
</li><br />
</p><br/><br/><br />
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<p><br />
<ul class="method"><br />
<li><a class="black-bold">A plant potentially able to release the produced pheromones into the environment.</a><br />
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<ul><br />
<li>Successful cloning of a trichome-specific promoter (PCPS2) from the genome of N tabacum and subsequent assembly with GFP as a reporter. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#tab2?display=block" class="normal-link-page">Results: Constructs-Pheromone release</a></li><br />
<li>Proof of the specificity of this promoter by GFP fluorescence detection. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/trichome_expression" class="normal-link-page">Results: Trichome-specific expression</a></li><br />
<li>Assembly of each gene of the pheromones production pathway with PCPS2 promoter and multigenic assembly with all three transcription units in a single plasmid. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#release" class="normal-link-page">Results: Constructs-Pheromone release</a></a></li><br />
</ul><br />
</li><br />
</p><br/><br/><br />
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<ul class="method"><br />
<li><a class="black-bold">A genetic switch able to control gene expression ready to be implemented in the plant. Results: Constructs-Switch</a><br />
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<ul><br />
<li>Cloning of the coding sequence from the CUP2 transcription factor from S cerevisiae and assembly with the CaMV constitutive promoter P35S. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#switch" class="normal-link-page">Results: Construct-Switch</a></li><br />
<li>Creation of the Cupper-responsive chimeric promoter and assembly with Firefly luciferase gene as a reporter, P19 as a gene silencing suppressor, and Renilla luciferase as control for Luciferase expression assay.<a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#switch" class="normal-link-page">Results: Construct-Switch</a></li><br />
<li>Multigenic assembly comprising the CUP2 transcriptional unit with the chimeric promoter and reporter gene assembly. <br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#switch" class="normal-link-page">Results: Construct-Switch</a></li><br />
</ul><br />
</li><br />
</p><br/><br/><br />
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<ul class="method"><br />
<li><a class="black-bold">A sterile and dark purple plant safe for living beings and the environment.<a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#switch" class="normal-link-page">Biosafety</a><br />
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<ul><br />
<li>Multigenic assembly of two biosafety devices, comprising the Barnase (male-sterility) with one chromoprotein in each device, AmilCP or AmilGFP (identity preservation). <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/constructs#biosafety" class="normal-link-page">Results: Constructs-Biosafety</a></li><br />
<li>Purple plant to preserve its identity expressing SlANT1 and SlJAF13 transcription factors. <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results" class="normal-link-page">Results: Biosafety</a></li><br />
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</ul><br />
</li><br />
</p><br/><br/><br />
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<p><br />
<a class="black-bold">Diffusion and communication of the project with involved experts and stakeholders.<a href="https://2014.igem.org/Team:Valencia_UPV/policy/activities" class="normal-link-page">Policy and Practices</a></a> <br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/results/biosafetyTeam:Valencia UPV/Project/results/biosafety2014-10-18T01:35:20Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results">Results</a> > <a>Biosafety</a></h3></p><br/><br/><br />
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<div align="center"><span class="coda"><roja>B</roja>iosafety</span> </div><br/><br/><br />
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<p>Our goal was to develop a sterile and easily identifiable plant. In order to do this, we created a module in collaboration with <a href="https://2014.igem.org/Team:NRP-UEA-Norwich" class="normal-link-page">NRP-UEA-Norwich</a> which incorporated an RNAse (barnase) under the regulation of a tapetum specific promoter (TA29) and a chromoprotein</p><br/><br />
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<p>Male Sterility strategy was not possible to test in a transient approach, but both components are well documented (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety">see Biosafety Module</a>).</p><br/><br />
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<p>The NRP-UEA-Norwich provided us with blue and yellow chromoproteins transcriptional units (TU) and we agroinfiltrated both TUs in <i>N. benthamiana</i> along with a GFP control. Chromoprotein detection was impossible by the naked eye (Figure 1) even though GFP control was expressed (Figure 2). Plants agroinfiltration was correct since GFP was expressed in the leaves.</p><br/><br/><br />
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<div align="center"><img width="500px" src="https://static.igem.org/mediawiki/2014/d/d6/VUPVFigure_1_Biosafety_Results.png"></img></div><br/><br />
<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1. Plants agroinfiltrated with NRP-UEA-Norwich chromoproteins</span>. TUs containing Blue (left) and Yellow (Right) chromoproteins were agroinfiltrated in N.benthamiana. None of the chromoproteins could be detected by the naked eye.</p></div><br/><br/><br />
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<div align="center"><img width="500px" src="https://static.igem.org/mediawiki/2014/7/74/VUPVFigure_2_Biosafety_Results.png"></img></div><br/><br />
<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 2. Agroinfiltration GFP control</span>. Agroinfiltration control shows GFP expression, indicating that non-detection of chromoproteins is not due to a failure in agroinfiltration.</p></div><br/><br/><br />
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<p>Nevertheless, identity preservation is an important part of the biosafety module, and it must be happen efficiently. <a href="https://2014.igem.org/Team:NRP-UEA-Norwich" class="normal-link-page">NRP-UEA-Norwich</a> team suggested leaf degreening in order to observe the chromoproteins, but we don’t consider this strategy convenient for our purpose since identity should be easily recognisable in plants without any kind of treatment. As an alternative to chromoproteins, we propose the use of two transcriptional factors to enhance anthocyanins production.</p><br/><br />
<br />
<p>Our identity preservation construct consists of two transcriptional units carrying the tomato (<i>Solanum lycopersicum</i>) transcriptional factors (SlANT1, SlJAF13) which are regulated by the 35S constitutive promoter. Both transcriptional factors are involved in flavonoids biosynthetic pathway regulation; it is that they enhance anthocyanin accumulation.</p><br/><br />
<br />
<p>SlANT1 and SlJAF13 are <i>S. lycopersicum</i> orthologous of the <i>Antirrhinum majus</i> Rosea1 and Delila genes. In previous researchs, ectopic over-expression of these transcription factors under the control of the E8 fruit-specific promoter increases the transcript levels of most of the anthocyanin biosynthetic genes in tomato fruit leading to high levels of <i>anthocyanins</i> [1,2]. In <i>N. benthamiana</i>, transient expression of SIANT1 and SIJAF3 activates the expression of several flavonoid biosynthetic genes leading to a change of the colour on the leaf due to accumulation of anthocyanins.</p><br/><br />
<br />
<p>We transiently transformed this new identity preservation module (containing SIANT1 and SIJAF13 transcriptional units) by agroinfiltration into <i>N. benthamiana</i>. As result, the anthocyanin accumulation drives our plant to a violet colour change that can be observed by the naked eye (Figure 3).</p><br/><br />
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<div align="center"><img width="500px" src="https://static.igem.org/mediawiki/2014/b/b8/VUPV_Rosea.png"></img></div><br/><br />
<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 3. Anthocyanins accumulation in <i>N.benthamiana</i></span>. SlANT1 and SlJAF13 expression produces accumulation of anthocyanins in agroinfiltrated plant (Left), showing a purple colour in contrast with a green wild type leaf (Right, held by hand).</p></div><br/><br/><br />
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<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/eag"><strong>&larr; Go to Electroantennography</strong></a><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/results/constructsTeam:Valencia UPV/Project/results/constructs2014-10-18T01:14:58Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results">Results</a> > <a>Constructs</a></h3></p><br/><br/><br />
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<div align="center"><span class="coda"><roja>C</roja>onstructs</span> </div><br/><br/><br />
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<a name="biosyn" class="subpart">RESULTS – CONSTRUCTS- BIOSYNTHESIS</a><br/><br/><br />
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<div><br />
<img title="Figure 1. Engineered pheromone production pathway" width="300px" style="float:right; margin-right: 15px;" src="https://static.igem.org/mediawiki/2014/0/05/VUPVPathway-semiochemical.jpg" alt="pathway_1"></img><br />
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<p>In order to engineer the <span class="blue-bold">insect sexual pheromone</span> <a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesis">pathway</a> in our Sexy plant, we had to isolate four genes from different organisms: a desaturase (AtrΔ11), a reductase (HarFAR), an acetyltransferase (EaDAcT) and finally an alcohol oxidase (FAO). As they were coming from very different and not easily accessible organisms (two moths and a plant from Asia), the coding sequences (CDS) of the first three enzymes were obtained by gene synthesis (Integrated DNA Technologies, IDT) after codon usage optimization for <i>N. benthamiana</i>. As for the fourth one (FAO) we tried to amplify it from the genomic DNA of the yeast <i>Candida tropicalis</i>, with no successful results. Nevertheless, the three synthetic genes were sufficient to produce at least two of our target pheromones, the alcohol <span class="red-bold">(Z)-11-hexadecen-1-ol</span> and the acetate <span class="blue-bold">(Z)-11-hexadecenyl acetate</span>.</p><br />
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<br/><p style="text-align: right; font-style: italic; font-size: 0.8em; width: 830px;"><span class="black-bold">Figure 1</span>. Engineered pheromone production pathway.</p></div><br/><br/><br />
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<p>All three DNA sequences were domesticated; that is, standardized as <a href="#" class="normal-link-page">GoldenBraid</a> parts and cloned in the pUPD plasmid. Subsequently, each CDS was assembled with the strong constitutive Cauliflower mosaic virus promoter (P35S) and its terminator (T35S) respectively, in a multipartite assembly reaction. P35S is a strong plant constitutive promoter with high expression levels. As result of these assemblies, we obtained three Transcriptional units (TU) ready for plant transformation.</p><br/><br />
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<div align="center"><br />
<img src="https://static.igem.org/mediawiki/2014/d/d5/VUPVPlasmido_Atr.png" alt="Atr-plasmid" width="250px"></img><br />
<img src="https://static.igem.org/mediawiki/2014/7/71/VUPVPlasmido_harfar-WIKI.png" alt="harfar_plasmid" width="250px"></img><br />
<img src="https://static.igem.org/mediawiki/2014/5/52/VUPVPlasmido_eadact-WIKI.png" alt="eadact_plasmid" width="250px"></img></div><br/><br />
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<p>To maximize the flow through of the pathway, we wanted to make sure that all three genes were co-delivered simultaneously. This is to ensure that each transformed cell receives a complete set of genes and that the expression of all three enzymes is balanced and coordinated. Co-delivery is achieved by creating a multigenic construct, where all three genes are assembled in a single plasmid.</p><br/><br />
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<p>We used the GoldenBraid assembly system to create the multigene assembly. After two binary reactions, the 3-genes construct was obtained. This construct was then transformed, by <span class="blue-bold">agroinfiltration</span>, into <i>N. benthamiana</i> plants for pheromone production.</p><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/b/bc/VUPVPlasmido_MULTI-WIKI.png" width="350px"></img></div><br/><br />
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<p>We used GoldenBraid 2.0 for all our cloning reactions. GoldenBraid and BioBrick parts are not directly exchangeable; however, we adapted the coding sequences of the three biosynthetic genes to BioBrick standards using a <a class="normal-link-page" href="#">GoldenBraid-Biobricks translator</a> developed by the <a href="https://2014.igem.org/Team:NRP-UEA-Norwich/Project_Mo-Flipper" class="normal-link-page">NRP-UEA-Norwich</a> team. These BioBricks have been submitted to the Parts Registry.</p><br/><br/><br/><br />
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<a name="phero" class="subpart">RESULTS-CONSTRUCTS-PHEROMONE RELEASE</a><br/><br/><br />
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<p>To obtain a plant capable of releasing pheromones into the environment in an efficient way, we decided to use the glandular trichome specific promoter (PCPS2) (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/release" class="normal-link-page">see Pheromone Relase</a>).</p><br/><br />
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<p>We obtained this promoter from Nicotiana tabacum genome and tested its functionality assembling it with GFP (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/trichome_expression" class="normal-link-page">see Results: Pheromone Release</a>).</p><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/8/8c/VUPVPlasmido_PCPS2-GFP.png" width="350px"></img></div><br/><br />
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<p>As it proved to be an effective promoter, expressing GFP only at the glandular cells of the trichomes, we decided to assemble each of the pheromone biosynthetic genes with this promoter to reach improvements in the pheromones release (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesis">see Biosynthesis</a>).</p><br/><br />
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<div align="center"><br />
<img src="https://static.igem.org/mediawiki/2014/0/09/VUPVPlasmido_Atr-PCPS.png" alt="Atr-plasmid" width="250px"></img><br />
<img src="https://static.igem.org/mediawiki/2014/f/f3/VUPVPlasmido_harfar-PCPS.png" alt="harfar_plasmid" width="250px"></img><br />
<img src="https://static.igem.org/mediawiki/2014/3/34/VUPVPlasmido_eadact-PCPS2.png" alt="eadact_plasmid" width="250px"></img></div><br/><br />
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<p>Finally, after two binary GoldenBraid assembly steps, we obtained a multigenic construct with all three transcriptional units with the PCPS2 promoter. This construct was ready to be transferred to the plant to test the release of the pheromones.</p><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/8/8b/VUPVPlasmido_MULTI-PCPS.png" width="350px"></img></div><br/><br/><br/><br />
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<a name="switch" class="subpart">RESULTS - CONSTRUCTS-SWITCH</a><br/><br/><br />
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<p>In order to have a tight control over the pheromone production in the plant, we implemented an inducible switch in the plant genetic circuit. This switch is induced with the presence of copper ions, activating the pheromone production pathway in the plant only when the insects mating season arrives. With this kind of inducible expression, the pheromones production metabolic cost for the plant will be reduced and the pheromone release will be always under control (<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/switch" class="normal-link-page">see Switch</a>).</p><br/><br />
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<p>This switch is composed of two different parts:</p><br/><br />
<ul style="list-style: inherit;"><br />
<li class="normal-sangría">A constituvely expressed transcription factor (CUP2) that changes its conformation in the presence of a given concentration of cupper ions. With this change of conformation, it can bind a specific promoter and initiates the transcription of a particular gene.</li><br />
<li class="normal-sangría">A chimeric promoter that includes an Upstream Activating Sequence (UAS) whith the transcription factor binding site.</li><br />
</ul><br/><br />
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<p>This switch is naturally present in yeast, so it must be adapted to plants.</p><br/><br />
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<p>Therefore, the transcription factor CUP2, which was obtained from S. cerevisae genome, was assembled with de Cauliflower mosaic virus (CaMV) P35S promoter and T35S terminator to be constitutively expressed. In addition, an activator domain (GAL4 AD) was joined to the CUP2 sequence, in order to improve the transcription initiation.</p><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/8/8a/VUPVPlasmido_CUP2.png" width="350px"></img></div><br/><br />
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<p>On the other hand, we needed to assemble the chimeric promoter and the gene of interest. This chimeric promoter is composed of 3 different parts:</p><br/><br />
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<ul style="list-style: inherit;"><br />
<li class="normal-sangría">The aforementioned UAS, which is a 44bp region. Only 16bp of them are actually the binding site for CUP2 while the rest are spacer nucleotides.</li><br />
<li class="normal-sangría">A minimal promoter, miniP35S (-60). These are a reduced number of nucleotides from the CaMV P35S promoter required for starting transcription.</li><br />
<li class="normal-sangría">The 5'-UTR region of the tobacco mosaic virus (TMV), called omega sequence. This sequence functions as a translational enhancer in plants.</li><br />
</ul><br/><br />
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<p>As we were going to perform a Luciferase assay to test the switch (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/results">see Luciferase assay</a>) the gene of interest assembled with the chimeric promoter was the Luciferase. This is the obtained construct.</p><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/c/cf/VUPVPlasmido_QUIMER.png" width="350px"></img></div><br/><br />
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<p>In order to have a control for the Luciferase assay, we needed another construct already available in the GoldenBraid collection. It included the Renilla and P19 genes.</p><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/8/84/VUPVPlasmido_renilla.png" width="350px"></img></div><br/><br />
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<p>Finally, we assembled all three constructs into a single multigenic construct that was introduced into the plant to perform the Luciferase assay.</p><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/b/b9/VUPVPlasmido_switch_completo.png" width="650px"></img></div><br/><br/><br/><br />
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<a name="biosafe" class="subpart">RESULTS-CONSTRUCTS-BIOSAFETY</a><br/><br/><br />
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<p>In order to obtain a safe plant we developed a biosafety module which avoids spread of the plant’s genetic material and allows easy identification of the modified plant.</p><br/><br />
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<p>In order to avoid spread of genetic material, we built a plasmid which constitutively expressed Barnase (an RNAse from <i>Bacillus amyloliquefaciens</i>) under the regulation of the tapetum specific promoter TA29. Barnase effect under the regulation of this promoter can’t be tested under the limitations of transient expression and time restrictions didn’t allow us to create the stable plant, but <a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety">its function is well documented</a>.</p><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/4/4e/VUPVFigure_1.png" width="350px"></img></div><br/><br />
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<p>We worked together with <a class="normal-link-page" href="https://2014.igem.org/Team:NRP-UEA-Norwich">NRP-UEA-Norwich</a> team to introduce their chromoproteins (Yellow and Blue) in our system to be used as a strategy of identity preservation. We received their complete transcriptional units and tested them in plant. However, their activity was not strong enough to be detected by naked eye.</p><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/6/6d/VUPVFigure_2.png" width="550px"></img></div><br/><br />
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<p>The complete biosafety module, containing chromoproteins as identity preservation agent, had already been constructed in biobricks format. Expression of Barnase specifically in tapetum cells and an identity preservation agent completes our biosafety module.</p><br/><br/><br />
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<div align="center"><img src="https://static.igem.org/mediawiki/2014/b/b3/VUPVFigure_3.png" width="550px"></img></div><br/><br />
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<p>Since the provided chromoproteins did not show enough change in the plant’s colour to be detected by the naked eye, we tested an alternative strategy. We tested an anthocyanin production enhancing genetic construction in <i>N.benthamiana</i>. Anthocyanins are coloured compounds found in plants.</p><br/><br />
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<p>We obtained this construction from <a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/gb">GoldenBraid 2.0</a> collection. This construction expresses two transcription factors (Ant1 and JAF13) and successfully enhanced the synthesis of anthocyanins. Therefore, we propose an identity preservation strategy based in anthocyanin accumulation as part of our biosafety module.</p><br/><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modulesTeam:Valencia UPV/Project/modules2014-10-18T01:12:56Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a>Project Modules</a></h3></p><br/><br />
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<div align="center"><span class="coda"><roja>P</roja>roject <roja>M</roja>odules</span> </div><br/><br/><br />
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<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/problem"><div class="modules" id="Problem"><br />
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<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/moths_behavior"><div class="modules" id="Moths_Behavior"><br />
</div></a><br />
</td><br />
<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesis"><div class="modules" id="Biosynthesis"><br />
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<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><div class="modules" id="Methodology"><br />
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</td><br />
<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety"><div class="modules" id="Biosafety"><br />
</div></a><br />
</td><br />
<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/ideal_plant"><div class="modules" id="Ideal_Plant"><br />
</div></a><br />
</td><br />
</tr><br />
<tr><br />
<td colspan="1"><br/><a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/overview"><strong>&larr; Go to Project Overview</strong></a></td><br />
<td colspan="2" align="center"><br/><a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Modeling/overview"><strong>Go to Modeling</strong></a></td><br />
<td colspan="1"><br/><a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results"><strong>Go to Results &rarr;</strong></a></td><br />
</tr><br />
</table><br />
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</div><br />
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<p style=" font-style: italic; font-size: 0.7em; width: 700px;"><br />
Sources of the original images:</p><br/><br />
<ul class="method" style="font-size: 0.5em;"><br />
<br />
<br />
<li>The Problem: <a class="normal-link-page" href="http://www.amanecerrural.com/campohoy/detalle.asp?idcontenido=19569">http://www.amanecerrural.com/campohoy/detalle.asp?idcontenido=19569</a></li><br/><br />
<br />
<li> Moths behaviour: <a class="normal-link-page" href="http://walterspropertyservices.com/the-invasive-gypsy-moth/">http://walterspropertyservices.com/the-invasive-gypsy-moth/</a> </li><br/><br />
<br />
<li>Pheromones biosynthesis: S. Schwizer & G. Martin, Boyce Thompson Institute.</li><br/><br />
<br />
<li>Pheromone Release: <a class="normal-link-page" href="http://www.cannabisculture.com/files/images/IMG_0218.jpg">http://www.cannabisculture.com/files/images/IMG_0218.jpg</a> </li><br/><br />
<br />
<li>Switch: <a class="normal-link-page" href="http://www.suwanee.com/pdfs/Organic%20Gardening%20PP%20022710.pdf">http://www.suwanee.com/pdfs/Organic%20Gardening%20PP%20022710.pdf</a> </li><br/><br />
<br />
<li>Methodology: <a class="normal-link-page" href="http://events.embo.org/12-plant-microbe/">http://events.embo.org/12-plant-microbe/</a> </li><br/><br />
<br />
<li>Biosafety: <a class="normal-link-page" href="http://xdesktopwallpapers.com/green-leave-closeup-macro-shot-22265.php">http://xdesktopwallpapers.com/green-leave-closeup-macro-shot-22265.php</a> </li><br/><br />
<br />
<li>Future perspectives: Ryan Kitko/Wikimedia Commons</li><br/></p><br />
</ul><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modulesTeam:Valencia UPV/Project/modules2014-10-18T01:11:08Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a>Project Modules</a></h3></p><br/><br />
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<div align="center"><span class="coda"><roja>P</roja>roject <roja>M</roja>odules</span> </div><br/><br/><br />
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<table align="center"><br />
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<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/problem"><div class="modules" id="Problem"><br />
</div></a><br />
</td><br />
<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/moths_behavior"><div class="modules" id="Moths_Behavior"><br />
</div></a><br />
</td><br />
<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesis"><div class="modules" id="Biosynthesis"><br />
</div></a><br />
</td><br />
<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/release"><div class="modules" id="Release"><br />
</div></a><br />
</td><br />
</tr><br />
<tr><br />
<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/switch"><div class="modules" id="Switch"><br />
</div></a><br />
</td><br />
<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><div class="modules" id="Methodology"><br />
</div></a><br />
</td><br />
<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety"><div class="modules" id="Biosafety"><br />
</div></a><br />
</td><br />
<td><br />
<a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/ideal_plant"><div class="modules" id="Ideal_Plant"><br />
</div></a><br />
</td><br />
</tr><br />
<tr><br />
<td colspan="1"><br/><a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/overview"><strong>&larr; Go to Project Overview</strong></a></td><br />
<td colspan="2" align="center"><br/><a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Modeling/overview"><strong>Go to Modeling</strong></a></td><br />
<td colspan="1"><br/><a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results"><strong>Go to Results &rarr;</strong></a></td><br />
</tr><br />
</table><br />
<br />
</div><br />
</br></br></div><br />
<div id="space-margin"></div><br />
<br />
<div align="center"><div id="cn-box" align="justify"><br />
<br />
<p style=" font-style: italic; font-size: 0.8em; width: 700px;"><br />
Sources of the original images:</p><br/><br />
<ul class="method" style="font-size: 0.5em;"><br />
<p style="text-align: center; font-style: italic; font-size: 0.8em; width: 700px;"><br />
<br />
<li>The Problem: <a class="normal-link-page" href="http://www.amanecerrural.com/campohoy/detalle.asp?idcontenido=19569">http://www.amanecerrural.com/campohoy/detalle.asp?idcontenido=19569</a></li><br/><br />
<br />
<li> Moths behaviour: <a class="normal-link-page" href="http://walterspropertyservices.com/the-invasive-gypsy-moth/">http://walterspropertyservices.com/the-invasive-gypsy-moth/</a> </li><br/><br />
<br />
<li>Pheromones biosynthesis: S. Schwizer & G. Martin, Boyce Thompson Institute.</li><br/><br />
<br />
<li>Pheromone Release: <a class="normal-link-page" href="http://www.cannabisculture.com/files/images/IMG_0218.jpg">http://www.cannabisculture.com/files/images/IMG_0218.jpg</a> </li><br/><br />
<br />
<li>Switch: <a class="normal-link-page" href="http://www.suwanee.com/pdfs/Organic%20Gardening%20PP%20022710.pdf">http://www.suwanee.com/pdfs/Organic%20Gardening%20PP%20022710.pdf</a> </li><br/><br />
<br />
<li>Methodology: <a class="normal-link-page" href="http://events.embo.org/12-plant-microbe/">http://events.embo.org/12-plant-microbe/</a> </li><br/><br />
<br />
<li>Biosafety: <a class="normal-link-page" href="http://xdesktopwallpapers.com/green-leave-closeup-macro-shot-22265.php">http://xdesktopwallpapers.com/green-leave-closeup-macro-shot-22265.php</a> </li><br/><br />
<br />
<li>Future perspectives: Ryan Kitko/Wikimedia Commons</li><br/></p><br />
</ul><br />
</div><br />
</html><br />
<br />
<br />
{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Team/AttributionsTeam:Valencia UPV/Team/Attributions2014-10-18T00:54:40Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Team</a> > <a>Attributions</a></h3></p><br/><br />
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<div align="center"><span class="coda"><roja>A</roja>ttributions</span> </div><br/><br/><br />
<!--<div align="center"><img alt="Attributions" src="https://static.igem.org/mediawiki/2014/1/13/VUPVAttributions.png"></img></div>--><br />
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<h3>Attributions in a flash</h3><br/><br />
<p><a name="Alba" class="encabezado">Alba Rubert:</a> Pheromone Analysis, Pheromone-Insect Interactions, BioBrick preparation, Policy and Practices.</p><br/><br />
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<p><a name="Alejandra" class="encabezado">Alejandra González:</a> Pheromone Diffusion modeling, Pheromone Production modeling, Policy and Practices.</p><br/><br />
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<br />
<p><a name="Alfredo" class="encabezado">Alfredo Quijano:</a> Pheromone Release, Switch, Pheromone Analysis, Pheromone-Insect Interactions, Policy and Practices.</p><br/><br />
<br />
<p><a name="Ivan" class="encabezado">Ivan Llopis:</a> Pheromone Production modeling, Wiki, Policy and Practices.</p><br/><br />
<br />
<br />
<p><a name="Jose" class="encabezado">José Gavaldá:</a> Pheromone Biosynthesis design, Pheromone Analysis, Policy and Practices</p><br/><br />
<br />
<br />
<p><a name="Lucia" class="encabezado">Lucía Estellés:</a> Pheromone Biosynthesis design, Biosafety, BioBrick preparation, Policy and Practices.</p><br/><br/><br/><br />
<br />
<br />
<h3>Attributions (Extended Version)</h3><br/><br />
<p><br />
<strong>Project: the idea.</strong><br />
We came to The Sexy Plant project after many brainstorming meetings including all the team. It would be impossible to define authors here: we all contributed to the design of the project.<br />
</p><br/><br />
<p><br />
<strong>Pheromone Biosynthesis.</strong><br />
Design of the biosynthesis pathway fell on <strong>Jose Gavaldá</strong> and <strong>Lucía Estellés</strong>. Pheromone expression analysis by GC-MS was carried out by <strong>Alba Rubert</strong>, <strong>Alfredo Quijano</strong>, <strong>Lucía Estellés</strong> and <strong>Jose Gavaldá</strong>. Pheromone-insect interaction analysis by means of wind tunnel and EAG was made by <strong>Alba Rubert</strong> and <strong>Alfredo Quijano</strong>.<br />
</p><br />
<p><br/><br />
<strong>Pheromone Release and Switch.</strong><br />
<strong>Alfredo Quijano</strong> was the responsible for the design and expression analysis of the trichome specific promoter. Copper inducible switch was designed by <strong>Alfredo Quijano</strong>. <br />
<br />
</p><br />
<p><br/><br />
<strong>Modeling.</strong><br />
<strong>Ivan Llopis</strong>, also supported by <strong>Alejandra González</strong>, worked in the FBA modeling to try to obtain estimation of the pheromone production and to try to optimize it. <strong>Alejandra González</strong> was in charge of the pheromone diffusion and moth behavior modeling to help decide where and how much sexy plants we should put in the field.<br />
<br />
</p><br/><br />
<p><br />
<strong>Biosafety.</strong><br />
Biosafety module was designed by <strong>Lucía Estellés</strong>, and Lucía was also the leader when fulfilling safety requirements: About our lab form (assisted by Alba, Alfredo and Jose), Check-in and Safety form.<br />
</p><br/><br />
<br />
<p><br />
<strong>Policy & Practices.</strong><br />
<strong>Lucía Estellés</strong>, <strong>Jose Gavaldá</strong>, <strong>Alejandra González</strong>, <strong>Alba Rubert</strong> and <strong>Alfredo Quijano</strong> dealed with the organization and execution of interviews with some stakeholders involved in the project. All the students in the team participated in the presentation of the Sexy Plant project to relevant members of the UPV, CSIC (Spanish Research Council) and Bayer CropScience business chair. <strong>Alba Rubert</strong>, <strong>Alfredo Quijano</strong>, <strong>Alejandra González</strong> and <strong>Ivan Llopis</strong> participated in the Generación Espontánea event. Also all the students organized and carry out the Summer School courses. And Lucía, Jose, Alba and Alfredo organized and participated in the lipdub.<br />
</p><br/><br />
<p><br />
<strong>BioBricks.</strong><br />
<strong>Lucía Estellés</strong>, helped by <strong>Alba Rubert</strong>, prepared the BioBricks for sending them to the Registry and documented the parts. <strong>Lucía</strong> was the designer of the Omega undercover part used to translate from GoldenBraid 2.0 to BioBricks standard.<br />
</p><br/><br />
<p><br />
<strong>Interlab Study.</strong><br />
<strong>Lucía Estellés</strong> performed the measurement interlab study.<br />
</p><br/><br />
<p><br />
<strong>Art and Design.</strong><br />
<strong>Alfredo Quijano</strong> was the major designer of the images used at the wiki, the presentation and the poster.<br />
</p><br/><br />
<p><br />
<strong>Wiki. </strong><br />
Performed by <strong>Ivan Llopis</strong>.<br />
</p><br/><br />
<p><br />
<strong>Poster.</strong><br />
<strong>Ivan Llopis</strong> worked in the poster design, all contributed with the concepts.<br />
<br />
</p><br/><br />
<p><br />
<strong>Talk.</strong> <strong>Lucía Estelles, Alfredo Quijano</strong> and <strong>Alejandra González.</strong> Also <strong>Alba Rubert</strong> performed Lucía's part when she was absent. All the team participated in the layout and conceptual flow. <br />
</p><br/><br/><br/><br />
<br />
<h3>Supervision</h3><br/><br />
<p><br />
All the Valencia UPV supervisors and advisors performed outstanding jobs. Their implication in the project is as follows:<br />
</p><br/><br />
<p><br />
<strong>Wetlab work</strong>. It was supervised by <strong>Esferanía Huet</strong> and <strong>Marta Vázquez</strong>. Always under the guidance and supervision of the wetlab director: <strong>Prof. Diego Orzáez.</strong><br />
</p><br/><br />
<p><br />
<strong>Modeling.</strong> FBA analysis was closely supervised by <strong>Yadira Boada</strong>, with the collaboration of <strong>Gabriel Bosque</strong> and <strong>Maria Siurana</strong>. Under the guidance of<strong> Profs. Javier Urchueguía and Jesús Picó. </strong>. Modeling of the Pheromone Diffusion and Moths response was supervised by <strong>Alejandro Vignoni</strong>. All under the guidance of<strong> Profs. Jesús Picó and Alberto Conejero. </strong><br />
</p><br/><br />
<p><br />
<strong>Wiki.</strong> Supervised by <strong>Victor Nina</strong> and <strong>Alejandro Vignoni</strong>.<br />
</p><br/><br />
<p><br />
<strong>Communication and logistics.</strong> <strong>Javier Urchieguía, Alberto Conejero, Maria Siurana, David Fuente</strong> and <strong>Yadira Boada</strong>.<br />
</p><br/><br />
<p><br />
<strong>Poster design and layout</strong> was supervised by<strong> Yadira Boada</strong>.<br />
</p><br/><br/><br/><br />
<br />
<h3>Acknowledgements</h3><br/><br />
<br />
<p>We thank the people who have helped in the project. We sincerely appreciate your contributions because <i>The Sexy Plant</i> project would not have been possible without your invaluable assistance and your enthusiasm: </p><br/><br />
<br />
<ul class="method"><br />
<br />
<li>Asun Fernández del Carmen, Institute for Plant Molecular and Cell Biology (IBMCP, CSIC-UPV): invaluable aid in everything. </li><br />
<li>Jose Luis Rambla Nebot, Institute for Plant Molecular and Cell Biology (IBMCP, CSIC-UPV): aid and advising in sample analysis by CG-MS and other technical advice. </li><br />
<li>Jaime Primo Millo, Ismael Navarro Fuertes, Vicente Navarro Llopis, Sandra Vacas González, Centre for Agricultural Chemical Ecology (CEQA - UPV): technical advice in project approach, insect management, electroantennography, wind tunnel, plant diffusion analysis, facility and technical details provision. </li><br />
<li>Christine Bäuerl, Institute of Automation and Industrial Computing ai2 - UPV: technical advice in the laboratory. </li><br />
<li>Jesús Muñoz Bertomeu, Institute for Plant Molecular and Cellular Biology (IBMCP, CSIC-UPV): <i>Candida tropicalis species</i> and assistance. </li><br />
<li>Lynne Yenush and Mª Carmen Marqués Romero, Institute for Plant Molecular and Cell Biology (IBMCP, CSIC-UPV): <i>Saccharomyces cerevisiae species</i>, genomic extraction protocols and reagents. </li><br />
<li>Alejandro Sarrión Perdigones, Baylor College of Medicine (Houston, Texas): technical advice and RFC development. </li><br />
<li>Matilde Eizaguirre Altuna, Department of Crop and Forest Sciences, University of Lleida (Lleida, Spain): <i>S. nonagrioides species</i> and useful information about moth handling. </li><br />
<li>NRP-UEA-Norwich team: MoFlippers and chromoproteins.. </li><br />
<li>IBMCP (CSIC-UPV) staff: Lipdub participation, hosting us at the Institute. </li><br />
<li>Silvia Michael: Logo design. </li><br />
<br />
</ul><br/><br />
<br />
<br />
<p>We would like to offer our special thanks to the following institutions for their support in the development of <i>The Sexy Plant Project</i>: </p><br/><br />
<br />
<ul class="method"><br />
<br />
<li>Universitat Politècnica de València (UPV) and Vice-rectorate for Student and University Extension, </li><br />
<li>Council of Valencia, </li><br />
<li>Bayer CropScience business chair, </li><br />
<li>Technical School of Industrial Engineering (ETSII - UPV), </li><br />
<li>Technical School of Agricultural Engineering (ETSIAMN - UPV) </li><br />
<br />
</ul><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_analysisTeam:Valencia UPV/Project/modules/methodology/sample analysis2014-10-18T00:51:43Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis GC-MS</a></h3></p><br/></br><br />
<br />
<div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis: <roja>G</roja>C-<roja>M</roja>S</span> </div><br/><br />
<br />
<p class="subpart">The Idea</p><br/><br />
<br />
<br />
<p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of the separation resolution provided by chromatography with the structural information provided by mass spectrometry allows the quantification and identification of each single volatile molecule present in the sample.</p><br/><br/><br />
<br />
<br />
<p class="subpart">GAS CROMATOGRAPHY</p><br/><br />
<br />
<p>As every chromatography technique, gas chromatography is based on differential partitioning of the components of a sample between a mobile phase that acts as sample carrier (a pure gas such as N2, He or H2 in the case of gas chromatography) and the stationary phase coating the chromatography column.</p><br/><br/><br />
<br />
<p>The molecules in the mixture are separated as they flow through the column by selective retention. Molecules with higher affinity for the mobile phase will flow faster and elute the column first, whereas those with higher affinity for the stationary phase will take longer to pass through the system. The retention time of a particular substance (the time it takes to pass through the column) depends on the type of column used, its length, and set temperature. </p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/4/47/VUPVGas_chromatography_1.jpg" alt="analytes"></img></div><br/><br />
<br />
<br />
<p>The last part of the chromatograph is the detector where a signal is registered as the compounds elute from the column. In GC-MS the mass spectrometer acts as detector.</p><br/><br/><br />
<br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/2/22/VUPVGas_chromatography_2.jpg" alt="sample_injector"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 1. </span>.Gas Chromatography diagram</p></div><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;">K. Murray/ Wikimedia Commons / CC-BY-SA-3.0.</p></div><br/><br/><br />
<br />
<p>Example of a chromatogram obtained by GC: each peak corresponds to a different molecule.</p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/b/bf/Gas_chromatography_3_2.jpg" alt="molecules_gc"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 2. </span>.Chromatogram obtained by Gas chromatography</p></div><br/><br/><br />
<br />
<br/><p class="subpart">MASS SPECTROMETRY</p><br/><br/><br />
<br />
<br />
<br />
<p>Mass spectrometry is an analytical technique capable of separating charged ions according to their mass-to-charge ratio (m/z) and measuring their abundance.</p><br/><br />
<p>The three basic components of a mass spectrometer are:</p><br />
<ul class="method"><br />
<br />
<li>The ion source, where ionization takes place. </li><br />
<li>The mass analyser.</li><br />
<li>The detector. </li><br />
<br />
</ul><br />
<br />
<p>MS systems differ in the methods used to generate and separate the ions. Our MS system employs electron ionization (EI), a quadrupole mass analyser, and an electron multiplier detector.<br />
</p><br/><br/><br />
<br />
<p><b>How it works</b><br />
</p><br/><br />
<br />
<ul class="method"><br />
<br />
<li><b>Ionization:</b> <br />
The different compounds in the sample mixture enter the ionization source as they elute form the column. There, they are bombarded with high-energy electrons (70eV), which break the molecules into charged fragments of a range of different masses, which are characteristic for each compound.<br />
</li><br/><br />
<li><b>Analysis:</b> <br />
The resulting fragments are separated according to their m/z ratio in the quadrupole analyser. A quadrupole consists of four cylindrical rods, two of them having positive electric potential while the other two are negatively charged. A radio frequency voltage is applied between the rod pairs creating an oscillating electric field. Only the ions with a given m/z will maintain its trajectory and cross the quadrupole to reach the detector, while the rest will be deflected. The voltage applied can be continuously changed (full scan) to monitor a range of m/z values, or it can be set to monitor only specific m/z ions (single ion monitoring mode, SIM)<br />
</li><br/><br />
<li><b>Detection:</b><br />
The ions that cross the analyzer reached the detector which converts ions to electric currency. The more quantity of ions that arrive, the greater the electron current produced. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal.<br />
</li><br/><br />
<br />
</ul><br />
<p>As a result a mass spectrum for each compound is obtained, i.e., the pattern of the ion fragments in which that compound breaks down, characterised by their m/z ratio and their relative abundance. This mass spectrum is characteristic for each substance and therefore a very valuable tool for compound identification.</p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/9/9e/VUPV_mass.png"></img></div><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 3. </span>(Z)-11-Hexadecn-1-ol mass spectrum</p></div><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Source</span>. NIST Chemistry Webbook</p></div><br/><br />
<br />
<br />
<br />
<br />
<p>You can find the results of the GC-MS analysis <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis" class="normal-link-page">here</a><br />
<p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/><br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation"><strong>&larr; Go to Sample Preparation</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><strong>Go back to Methodology</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/dynamic_headspace"><strong>Go to Sampling Technique &rarr;</strong></a></div></br></br></br><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_analysisTeam:Valencia UPV/Project/modules/methodology/sample analysis2014-10-18T00:51:17Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis GC-MS</a></h3></p><br/></br><br />
<br />
<div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis: <roja>G</roja>C-<roja>M</roja>S</span> </div><br/><br />
<br />
<p class="subpart">The Idea</p><br/><br />
<br />
<br />
<p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of the separation resolution provided by chromatography with the structural information provided by mass spectrometry allows the quantification and identification of each single volatile molecule present in the sample.</p><br/><br/><br />
<br />
<br />
<p class="subpart">GAS CROMATOGRAPHY</p><br/><br />
<br />
<p>As every chromatography technique, gas chromatography is based on differential partitioning of the components of a sample between a mobile phase that acts as sample carrier (a pure gas such as N2, He or H2 in the case of gas chromatography) and the stationary phase coating the chromatography column.</p><br/><br/><br />
<br />
<p>The molecules in the mixture are separated as they flow through the column by selective retention. Molecules with higher affinity for the mobile phase will flow faster and elute the column first, whereas those with higher affinity for the stationary phase will take longer to pass through the system. The retention time of a particular substance (the time it takes to pass through the column) depends on the type of column used, its length, and set temperature. </p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/4/47/VUPVGas_chromatography_1.jpg" alt="analytes"></img></div><br/><br />
<br />
<br />
<p>The last part of the chromatograph is the detector where a signal is registered as the compounds elute from the column. In GC-MS the mass spectrometer acts as detector.</p><br/><br/><br />
<br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/2/22/VUPVGas_chromatography_2.jpg" alt="sample_injector"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 1. </span>.Gas Chromatography diagram</p></div><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;">K. Murray/ Wikimedia Commons / CC-BY-SA-3.0.</p></div><br/><br/><br />
<br />
<p>Example of a chromatogram obtained by GC: each peak corresponds to a different molecule.</p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/b/bf/Gas_chromatography_3_2.jpg" alt="molecules_gc"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 2. </span>.Chromatogram obtained by Gas chromatography</p></div><br/><br/><br />
<br />
<br/><p class="subpart">MASS SPECTROMETRY</p><br/><br/><br />
<br />
<br />
<br />
<p>Mass spectrometry is an analytical technique capable of separating charged ions according to their mass-to-charge ratio (m/z) and measuring their abundance.</p><br/><br />
<p>The three basic components of a mass spectrometer are:</p><br />
<ul class="method"><br />
<br />
<li>The ion source, where ionization takes place. </li><br />
<li>The mass analyser.</li><br />
<li>The detector. </li><br />
<br />
</ul><br />
<br />
<p>MS systems differ in the methods used to generate and separate the ions. Our MS system employs electron ionization (EI), a quadrupole mass analyser, and an electron multiplier detector.<br />
</p><br/><br/><br />
<br />
<p><b>How it works</b><br />
</p><br/><br />
<br />
<ul class="method"><br />
<br />
<li><b>Ionization:</b> <br />
The different compounds in the sample mixture enter the ionization source as they elute form the column. There, they are bombarded with high-energy electrons (70eV), which break the molecules into charged fragments of a range of different masses, which are characteristic for each compound.<br />
</li><br/><br />
<li><b>Analysis:</b> <br />
The resulting fragments are separated according to their m/z ratio in the quadrupole analyser. A quadrupole consists of four cylindrical rods, two of them having positive electric potential while the other two are negatively charged. A radio frequency voltage is applied between the rod pairs creating an oscillating electric field. Only the ions with a given m/z will maintain its trajectory and cross the quadrupole to reach the detector, while the rest will be deflected. The voltage applied can be continuously changed (full scan) to monitor a range of m/z values, or it can be set to monitor only specific m/z ions (single ion monitoring mode, SIM)<br />
</li><br/><br />
<li><b>Detection:</b><br />
The ions that cross the analyzer reached the detector which converts ions to electric currency. The more quantity of ions that arrive, the greater the electron current produced. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal.<br />
</li><br/><br />
<br />
</ul><br />
<p>As a result a mass spectrum for each compound is obtained, i.e., the pattern of the ion fragments in which that compound breaks down, characterised by their m/z ratio and their relative abundance. This mass spectrum is characteristic for each substance and therefore a very valuable tool for compound identification.</p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/9/9e/VUPV_mass.png"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 3. </span>(Z)-11-Hexadecn-1-ol mass spectrum</p></div><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Source</span>. NIST Chemistry Webbook</p></div><br/><br />
<br />
<br />
<br />
<br />
<p>You can find the results of the GC-MS analysis <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis" class="normal-link-page">here</a><br />
<p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/><br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation"><strong>&larr; Go to Sample Preparation</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><strong>Go back to Methodology</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/dynamic_headspace"><strong>Go to Sampling Technique &rarr;</strong></a></div></br></br></br><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_analysisTeam:Valencia UPV/Project/modules/methodology/sample analysis2014-10-18T00:50:43Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis GC-MS</a></h3></p><br/></br><br />
<br />
<div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis: <roja>G</roja>C-<roja>M</roja>S</span> </div><br/><br />
<br />
<p class="subpart">The Idea</p><br/><br />
<br />
<br />
<p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of the separation resolution provided by chromatography with the structural information provided by mass spectrometry allows the quantification and identification of each single volatile molecule present in the sample.</p><br/><br/><br />
<br />
<br />
<p class="subpart">GAS CROMATOGRAPHY</p><br/><br />
<br />
<p>As every chromatography technique, gas chromatography is based on differential partitioning of the components of a sample between a mobile phase that acts as sample carrier (a pure gas such as N2, He or H2 in the case of gas chromatography) and the stationary phase coating the chromatography column.</p><br/><br/><br />
<br />
<p>The molecules in the mixture are separated as they flow through the column by selective retention. Molecules with higher affinity for the mobile phase will flow faster and elute the column first, whereas those with higher affinity for the stationary phase will take longer to pass through the system. The retention time of a particular substance (the time it takes to pass through the column) depends on the type of column used, its length, and set temperature. </p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/4/47/VUPVGas_chromatography_1.jpg" alt="analytes"></img></div><br/><br />
<br />
<br />
<p>The last part of the chromatograph is the detector where a signal is registered as the compounds elute from the column. In GC-MS the mass spectrometer acts as detector.</p><br/><br/><br />
<br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/2/22/VUPVGas_chromatography_2.jpg" alt="sample_injector"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 1. </span>.Gas Chromatography diagram</p></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;">K. Murray/ Wikimedia Commons / CC-BY-SA-3.0.</p></div><br/><br />
<br />
<p>Example of a chromatogram obtained by GC: each peak corresponds to a different molecule.</p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/b/bf/Gas_chromatography_3_2.jpg" alt="molecules_gc"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 2. </span>.Chromatogram obtained by Gas chromatography</p></div><br/><br/><br />
<br />
<br/><p class="subpart">MASS SPECTROMETRY</p><br/><br/><br />
<br />
<br />
<br />
<p>Mass spectrometry is an analytical technique capable of separating charged ions according to their mass-to-charge ratio (m/z) and measuring their abundance.</p><br/><br />
<p>The three basic components of a mass spectrometer are:</p><br />
<ul class="method"><br />
<br />
<li>The ion source, where ionization takes place. </li><br />
<li>The mass analyser.</li><br />
<li>The detector. </li><br />
<br />
</ul><br />
<br />
<p>MS systems differ in the methods used to generate and separate the ions. Our MS system employs electron ionization (EI), a quadrupole mass analyser, and an electron multiplier detector.<br />
</p><br/><br/><br />
<br />
<p><b>How it works</b><br />
</p><br/><br />
<br />
<ul class="method"><br />
<br />
<li><b>Ionization:</b> <br />
The different compounds in the sample mixture enter the ionization source as they elute form the column. There, they are bombarded with high-energy electrons (70eV), which break the molecules into charged fragments of a range of different masses, which are characteristic for each compound.<br />
</li><br/><br />
<li><b>Analysis:</b> <br />
The resulting fragments are separated according to their m/z ratio in the quadrupole analyser. A quadrupole consists of four cylindrical rods, two of them having positive electric potential while the other two are negatively charged. A radio frequency voltage is applied between the rod pairs creating an oscillating electric field. Only the ions with a given m/z will maintain its trajectory and cross the quadrupole to reach the detector, while the rest will be deflected. The voltage applied can be continuously changed (full scan) to monitor a range of m/z values, or it can be set to monitor only specific m/z ions (single ion monitoring mode, SIM)<br />
</li><br/><br />
<li><b>Detection:</b><br />
The ions that cross the analyzer reached the detector which converts ions to electric currency. The more quantity of ions that arrive, the greater the electron current produced. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal.<br />
</li><br/><br />
<br />
</ul><br />
<p>As a result a mass spectrum for each compound is obtained, i.e., the pattern of the ion fragments in which that compound breaks down, characterised by their m/z ratio and their relative abundance. This mass spectrum is characteristic for each substance and therefore a very valuable tool for compound identification.</p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/9/9e/VUPV_mass.png"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 3. </span>(Z)-11-Hexadecn-1-ol mass spectrum</p></div><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Source</span>. NIST Chemistry Webbook</p></div><br/><br />
<br />
<br />
<br />
<br />
<p>You can find the results of the GC-MS analysis <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis" class="normal-link-page">here</a><br />
<p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/><br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation"><strong>&larr; Go to Sample Preparation</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><strong>Go back to Methodology</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/dynamic_headspace"><strong>Go to Sampling Technique &rarr;</strong></a></div></br></br></br><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_analysisTeam:Valencia UPV/Project/modules/methodology/sample analysis2014-10-18T00:48:30Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis GC-MS</a></h3></p><br/></br><br />
<br />
<div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis: <roja>G</roja>C-<roja>M</roja>S</span> </div><br/><br />
<br />
<p class="subpart">The Idea</p><br/><br />
<br />
<br />
<p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of the separation resolution provided by chromatography with the structural information provided by mass spectrometry allows the quantification and identification of each single volatile molecule present in the sample.</p><br/><br/><br />
<br />
<br />
<p class="subpart">GAS CROMATOGRAPHY</p><br/><br />
<br />
<p>As every chromatography technique, gas chromatography is based on differential partitioning of the components of a sample between a mobile phase that acts as sample carrier (a pure gas such as N2, He or H2 in the case of gas chromatography) and the stationary phase coating the chromatography column.</p><br/><br/><br />
<br />
<p>The molecules in the mixture are separated as they flow through the column by selective retention. Molecules with higher affinity for the mobile phase will flow faster and elute the column first, whereas those with higher affinity for the stationary phase will take longer to pass through the system. The retention time of a particular substance (the time it takes to pass through the column) depends on the type of column used, its length, and set temperature. </p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/4/47/VUPVGas_chromatography_1.jpg" alt="analytes"></img></div><br/><br />
<br />
<br />
<p>The last part of the chromatograph is the detector where a signal is registered as the compounds elute from the column. In GC-MS the mass spectrometer acts as detector.</p><br/><br/><br />
<br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/2/22/VUPVGas_chromatography_2.jpg" alt="sample_injector"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 1. </span>.Gas Chromatography diagram</p></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;">K. Murray/ Wikimedia Commons / CC-BY-SA-3.0.</p></div><br/><br />
<br />
<p>Example of a chromatogram obtained by GC: each peak corresponds to a different molecule.</p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/b/bf/Gas_chromatography_3_2.jpg" alt="molecules_gc"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 2. </span>.Chromatogram obtained by Gas chromatography</p></div><br/><br/><br />
<br />
<br/><p class="subpart">MASS SPECTROMETRY</p><br/><br/><br />
<br />
<br />
<br />
<p>Mass spectrometry is an analytical technique capable of separating charged ions according to their mass-to-charge ratio (m/z) and measuring their abundance.</p><br/><br/><br />
<p>The three basic components of a mass spectrometer are:</p><br />
<ul class="method"><br />
<br />
<li>The ion source, where ionization takes place. </li><br />
<li>The mass analyser.</li><br />
<li>The detector. </li><br />
<br />
</ul><br />
<br />
<p>MS systems differ in the methods used to generate and separate the ions. Our MS system employs electron ionization (EI), a quadrupole mass analyser, and an electron multiplier detector.<br />
</p><br/><br/><br />
<br />
<p><b>How it works</b><br />
</p><br/><br />
<br />
<ul class="method"><br />
<br />
<li><b>Ionization:</b> <br />
The different compounds in the sample mixture enter the ionization source as they elute form the column. There, they are bombarded with high-energy electrons (70eV), which break the molecules into charged fragments of a range of different masses, which are characteristic for each compound.<br />
</li><br/><br />
<li><b>Analysis:</b> <br />
The resulting fragments are separated according to their m/z ratio in the quadrupole analyser. A quadrupole consists of four cylindrical rods, two of them having positive electric potential while the other two are negatively charged. A radio frequency voltage is applied between the rod pairs creating an oscillating electric field. Only the ions with a given m/z will maintain its trajectory and cross the quadrupole to reach the detector, while the rest will be deflected. The voltage applied can be continuously changed (full scan) to monitor a range of m/z values, or it can be set to monitor only specific m/z ions (single ion monitoring mode, SIM)<br />
</li><br/><br />
<li><b>Detection:</b><br />
The ions that cross the analyzer reached the detector which converts ions to electric currency. The more quantity of ions that arrive, the greater the electron current produced. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal.<br />
</li><br/><br />
<br />
</ul><br />
<p>As a result a mass spectrum for each compound is obtained, i.e., the pattern of the ion fragments in which that compound breaks down, characterised by their m/z ratio and their relative abundance. This mass spectrum is characteristic for each substance and therefore a very valuable tool for compound identification.</p><br/><br/><br />
<br />
<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/9/9e/VUPV_mass.png"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 3. </span>(Z)-11-Hexadecn-1-ol mass spectrum</p></div><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Source</span>. NIST Chemistry Webbook</p></div><br/><br />
<br />
<br />
<br />
<br />
<p>You can find the results of the GC-MS analysis <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis" class="normal-link-page">here</a><br />
<p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/><br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation"><strong>&larr; Go to Sample Preparation</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology"><strong>Go back to Methodology</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/dynamic_headspace"><strong>Go to Sampling Technique &rarr;</strong></a></div></br></br></br><br />
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology">Methodology</a> > <a>Sample Analysis GC-MS</a></h3></p><br/></br><br />
<br />
<div align="center"><span class="coda"><roja>S</roja>ample <roja>A</roja>nalysis. <roja>G</roja>C-<roja>M</roja>S</span> </div><br/><br />
<br />
<p class="subpart">The Idea</p><br/><br />
<br />
<br />
<p>When it comes to analysing volatile compounds Gas Chromatography (GC) coupled to Mass spectrometry (MS) is unequivocally the first choice. The combination of the separation resolution provided by chromatography with the structural information provided by mass spectrometry allows the quantification and identification of each single volatile molecule present in the sample.</p><br/><br/><br />
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<br />
<p class="subpart">GAS CROMATOGRAPHY</p><br/><br />
<br />
<p>As every chromatography technique, gas chromatography is based on differential partitioning of the components of a sample between a mobile phase that acts as sample carrier (a pure gas such as N2, He or H2 in the case of gas chromatography) and the stationary phase coating the chromatography column.</p><br/><br/><br />
<br />
<p>The molecules in the mixture are separated as they flow through the column by selective retention. Molecules with higher affinity for the mobile phase will flow faster and elute the column first, whereas those with higher affinity for the stationary phase will take longer to pass through the system. The retention time of a particular substance (the time it takes to pass through the column) depends on the type of column used, its length, and set temperature. </p><br/><br/><br />
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<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/4/47/VUPVGas_chromatography_1.jpg" alt="analytes"></img></div><br/><br />
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<br />
<p>The last part of the chromatograph is the detector where a signal is registered as the compounds elute from the column. In GC-MS the mass spectrometer acts as detector.</p><br/><br/><br />
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<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/2/22/VUPVGas_chromatography_2.jpg" alt="sample_injector"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 1. </span>.Gas Chromatography diagram</p></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;">K. Murray/ Wikimedia Commons / CC-BY-SA-3.0.</p></div><br/><br />
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<p>Example of a chromatogram obtained by GC: each peak corresponds to a different molecule.</p><br/><br/><br />
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<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/b/bf/Gas_chromatography_3_2.jpg" alt="molecules_gc"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 2. </span>.Chromatogram obtained by Gas chromatography</p></div><br/><br/><br />
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<br/><p class="subpart">MASS SPECTROMETRY</p><br/><br/><br />
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<br />
<br />
<p>Mass spectrometry is an analytical technique capable of separating charged ions according to their mass-to-charge ratio (m/z) and measuring their abundance.</p><br/><br/><br />
<p>The three basic components of a mass spectrometer are:</p><br />
<ul class="method"><br />
<br />
<li>The ion source, where ionization takes place. </li><br />
<li>The mass analyser.</li><br />
<li>The detector. </li><br />
<br />
</ul><br />
<br />
<p>MS systems differ in the methods used to generate and separate the ions. Our MS system employs electron ionization (EI), a quadrupole mass analyser, and an electron multiplier detector.<br />
</p><br/><br/><br />
<br />
<p><b>How it works</b><br />
</p><br/><br />
<br />
<ul class="method"><br />
<br />
<li><b>Ionization:</b> <br />
The different compounds in the sample mixture enter the ionization source as they elute form the column. There, they are bombarded with high-energy electrons (70eV), which break the molecules into charged fragments of a range of different masses, which are characteristic for each compound.<br />
</li><br/><br />
<li><b>Analysis:</b> <br />
The resulting fragments are separated according to their m/z ratio in the quadrupole analyser. A quadrupole consists of four cylindrical rods, two of them having positive electric potential while the other two are negatively charged. A radio frequency voltage is applied between the rod pairs creating an oscillating electric field. Only the ions with a given m/z will maintain its trajectory and cross the quadrupole to reach the detector, while the rest will be deflected. The voltage applied can be continuously changed (full scan) to monitor a range of m/z values, or it can be set to monitor only specific m/z ions (single ion monitoring mode, SIM)<br />
</li><br/><br />
<li><b>Detection:</b><br />
The ions that cross the analyzer reached the detector which converts ions to electric currency. The more quantity of ions that arrive, the greater the electron current produced. Therefore, the system is capable of quantifying the arriving ions by measuring the produced electric signal.<br />
</li><br/><br />
<br />
</ul><br />
<p>As a result a mass spectrum for each compound is obtained, i.e., the pattern of the ion fragments in which that compound breaks down, characterised by their m/z ratio and their relative abundance. This mass spectrum is characteristic for each substance and therefore a very valuable tool for compound identification.</p><br/><br/><br />
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<div align="center"><img width="600px" src="https://static.igem.org/mediawiki/2014/9/9e/VUPV_mass.png"></img></div><br/><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Figure 3. </span>(Z)-11-Hexadecn-1-ol mass spectrum</p></div><br />
<div align="center"><p style="font-size: 0.8em; width: 70%;"><span class="black-bold">Source</span>. NIST Chemistry Webbook</p></div><br/><br />
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<p>You can find the results of the GC-MS analysis <a hrf=https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis> here.</a></a></p><br/><br />
<p>To see more details about GC-MS conditions <a class="blue-bold">see Protocol</a>.</p><br/><br/><br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/sample_preparation"><strong>&larr; Go to Sample Preparation</strong></a><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/File:VUPV_mass.pngFile:VUPV mass.png2014-10-18T00:18:34Z<p>Alquiru: </p>
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<div></div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/eagTeam:Valencia UPV/Project/eag2014-10-17T23:38:45Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results">Results</a> > <a>Electroantennography</a></h3></p><br/><br/><br />
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<div align="center"><span class="coda"><roja>E</roja>lectroantennography</span> </div><br/><br/><br />
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<div><br />
<img width="300px" style="float:right; margin-left: 15px;" src="https://static.igem.org/mediawiki/2014/f/f3/VUPVIMG_20141006_135403.jpg" alt="EAG_1"></img><br />
<p>As explained in the methodology section (<a href="#" class="normal-link-page">see Methodology: Electroantennography</a>) we performed an electroantennography (EAG) to test the moth response to pheromones. Insects can detect pheromones through their antennae, then an electrical impulse is transmitted from them to the brain in order to trigger moth response to the pheromones. The EAG allows us to detect these electrical impulses by connecting one insect antenna to two electrodes that will amplify this impulse in order to be detected.</p><br/><br/><br />
<br />
<p>We connected one antenna from a male moth, Sesamia nonagrioides , with the two electrodes. Then , an air current with a leaf extract containing our pheromones was applied (Figure 3. Signal 1). As it can be appreciated, as the extract was applied the antenna transmitted an electrical impulse. This was the moth response to our insect pheromones produced in plant.</p><br/><br />
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<br/><p style="text-align: right; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1</span>. Side view of an insect antenna connected to EAG electrodes.</p><br/><br />
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</div><br />
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<div><img style="float:left; margin-right: 15px;" width="300px" src="https://static.igem.org/mediawiki/2014/6/69/VUPVIMG_4156.JPG" alt="EAG_2"></img></div><br/><br />
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<br />
<p><br/><br/>As a control, we also applied an air current with no pheromones in suspension. (Figure 3. Signal 2) The antena did not transmit any electrical signal.</p><br/><br />
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<br/><p style="text-align: right; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 2</span>. Upper view of an insect antenna connected to EAG electrodes .</p><br/><br />
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<br />
<div align="center"><img width="500px" src="https://static.igem.org/mediawiki/2014/5/53/VUPVEAGi.png" alt="EAG"></img></div><br/><br />
<div align="center"><p style="text-align: center; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 3</span>. Electroantennography analysis of Sesamia nonagroides response to sexual pheromones produced in genetically engineered Nicotiana Benthamiana plants. Signal1: Antennal response to the Sexy Plant leaf extract. Signal 2: Antennal response to an air puff.</p></div><br/><br />
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<p>The volatiles in our Sexy Plants induced detectable electric pulses that could indicate a pheromone response, although further testing will be required for confirmation.</p><br/><br/><br />
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<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results/pheromone_analysis"><strong>&larr; Go to Pheromone Analysis</strong></a><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/File:VUPVEAGi.pngFile:VUPVEAGi.png2014-10-17T23:26:58Z<p>Alquiru: uploaded a new version of &quot;File:VUPVEAGi.png&quot;: Reverted to version as of 23:23, 17 October 2014</p>
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<div></div>Alquiruhttp://2014.igem.org/File:VUPVEAGi.pngFile:VUPVEAGi.png2014-10-17T23:23:28Z<p>Alquiru: uploaded a new version of &quot;File:VUPVEAGi.png&quot;</p>
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<div></div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/overviewTeam:Valencia UPV/Project/overview2014-10-17T22:42:19Z<p>Alquiru: </p>
<hr />
<div>{{:Team:Valencia_UPV/header}}<br />
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<div align="center"><div id="cn-box" align="justify"><br />
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<p><h3 class="hook" align="left"><a>Project</a> > <a>Overview</a></h3></p><br/><br />
<br />
<div align="center"><span class="coda"><roja>P</roja>roject <roja>O</roja>verview</span> </div><br/><br/><br />
<br />
<p>Pests cause <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/problem" class="normal-link-page">great economical loses</a> in agriculture and impede an optimal use of the resources. <span class="red-bold">Sexy Plant</span> rises as a pest control strategy based on the use of <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/moths_behavior" class="normal-link-page">mating disruption</a> by sex pheromones. The release of sex pheromones impedes male moths to find their females, avoiding crop-damaging larvae to be born.</p><br/><br />
<br />
<p>Sexy Plant is engineered to <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesis" class="normal-link-page">produce</a> and <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/release" class="normal-link-page">release</a> three moth sex pheromones which affect a broad number of Lepidoptera species (moths). We implemented previous work by Ding et al. [1] to produce the first two pheromones (<span class="red-bold">Z11-16:OH</span> and <span class="blue-bold">Z11-16:OAc</span>). Inspired by Hagström et al [2], we planned the introduction of a Fatty Acid reductase to transform <span class="red-bold">Z11-16:OH</span> to <span class="green-bold">Z11-16:Ald</span>. As result, the sexy plant will be able to produce three sex pheromones involved in moth’s mating disruption (Figure 1).</p><br/><br />
<br />
<div align="center"><img src="https://static.igem.org/mediawiki/2014/4/43/VUPVOv_fig2.png" alt="pheromone_pathway" title="Pheromone Pathway" width="450px"></img></div><br/><br />
<div align="center"><p style="text-align: center; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1. Biosynthetic pathway of moth sex pheromones</span>. Sex pheromones are bordered in purple.</p></div><br/><br />
<br />
<br />
<p>Biosafety is a major concern in our project. For that reason, we have developed a <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety" class="normal-link-page">biosafety module</a> which allows an easy identification of the plant by identity preservation and prevents the plant to spread its genetic material via pollen.</p><br/><br />
<br />
<div align="center"><img width="400px" src="https://static.igem.org/mediawiki/2014/6/64/VUPVOv_fig1.png"></img></div><br/><br />
<br />
<p>In addition, we decided to enable external control of pheromone production, designing a <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/switch" class="normal-link-page">genetic switch</a> that controls the activation of the biosynthetic pathway. This genetic switch turns on the production of pheromones when a solution containing CuSO4 is sprayed on the plant.</p><br/><br />
<br />
<br />
<p>The different genetic modules are put together to create the complete system. The system resembles an electric circuit, as it is represented in figure 2. Our genetic circuit, once introduced in <i>Nicotiana benthamiana</i> creates the <span class="red-bold">Sexy Plant</span>, a plant to fight moths and avoid damages in crops.</p><br/><br />
<br />
<br />
<div align="center"><img width="400px" src="https://static.igem.org/mediawiki/2014/a/ac/VUPVOverview_figure_2.png" alt="circuit"></img></div><br/><br />
<br />
<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 2. Our system in an electric circuit-like format</span>. The plant’s own metabolism is used as the energy source. The biosafety module, composed by identity preservation and male sterility parts is always under expression in the system, to ensure the security of the transgenic plant. Pheromone synthetic pathways (A,B or C, depending on the problem pest) are activated using a genetic switch sensitive to Copper Sulfate, so system will not be producing pheromones unless it is activated with Copper-rich solution.</p></div><br/><br/><br/><br/><br/><br/><br />
<br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV"><strong>&larr; Go to Home</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules"><strong>Go to Modules</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results"><strong>Go to Results &rarr;</strong></a></div><br/><br/><br/><br/><br />
<br />
<p align="center" class="black-bold">REFERENCES</p><br/><br />
<div style="position: relative; left: 3%; width: 96%;"><ol><br />
<li>Ding BJ, Hofvander P, Wang HL, Durrett TP, Stymne S, et al. (2014) A plant factory for moth pheromone production. Nat Commun 5: 3353.</li><br />
<li>Hagström A, Wang HL, Lienard MA, Lassance JM, Johansson T, et al. (2013) A moth pheromone brewery: production of (Z)-11-hexadecenol by heterologous co-expression of two biosynthetic genes from a noctuid moth in a yeast cell factory. Microb Cell Fact 12: 125.</li><br />
</ol></div><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/overviewTeam:Valencia UPV/Project/overview2014-10-17T22:41:32Z<p>Alquiru: </p>
<hr />
<div>{{:Team:Valencia_UPV/header}}<br />
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<br />
<p><h3 class="hook" align="left"><a>Project</a> > <a>Overview</a></h3></p><br/><br />
<br />
<div align="center"><span class="coda"><roja>P</roja>roject <roja>O</roja>verview</span> </div><br/><br/><br />
<br />
<p>Pests cause <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/problem" class="normal-link-page">great economical loses</a> in agriculture and impede an optimal use of the resources. <span class="red-bold">Sexy Plant</span> rises as a pest control strategy based on the use of <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/moths_behavior" class="normal-link-page">mating disruption</a> by sex pheromones. The release of sex pheromones impedes male moths to find their females, avoiding crop-damaging larvae to be born.</p><br/><br />
<br />
<p>Sexy Plant is engineered to <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesis" class="normal-link-page">produce</a> and <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/release" class="normal-link-page">release</a> three moth sex pheromones which affect a broad number of Lepidoptera species (moths). We implemented previous work by Ding et al. [1] to produce the first two pheromones (<span class="red-bold">Z11-16:OH</span> and <span class="blue-bold">Z11-16:OAc</span>). Inspired by Hagström et al [2], we planned the introduction of a Fatty Acid reductase to transform <span class="red-bold">Z11-16:OH</span> to <span class="green-bold">Z11-16:Ald</span>. As result, the sexy plant will be able to produce three sex pheromones involved in moth’s mating disruption (Figure 1).</p><br/><br />
<br />
<div align="center"><img src="https://static.igem.org/mediawiki/2014/4/43/VUPVOv_fig2.png" alt="pheromone_pathway" title="Pheromone Pathway" width="450px"></img></div><br/><br />
<div align="center"><p style="text-align: center; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1. Biosynthetic pathway of moth sex pheromones</span>. Sex pheromones are bordered in purple.</p></div><br/><br />
<br />
<br />
<p>Biosafety is a major concern in our project. For that reason, we have developed a <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety" class="normal-link-page">biosafety module</a> which allows an easy identification of the plant by identity preservation and prevents the plant to spread its genetic material via pollen.</p><br/><br />
<br />
<div align="center"><img width="400px" src="https://static.igem.org/mediawiki/2014/6/64/VUPVOv_fig1.png"></img></div><br/><br />
<br />
<p>In addition, we decided to enable external control of pheromone production, designing a <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/switch" class="normal-link-page">genetic switch</a> that controls the activation of the biosynthetic pathway. This genetic switch turns on the production of pheromones when a solution containing CuSO4 is sprayed on the plant.</p><br/><br />
<br />
<br />
<p>The different genetic modules are put together to create the complete system. The system resembles an electric circuit, as it is represented in figure 2. Our genetic circuit, once introduced in <i>Nicotiana benthamiana</i> creates the Sexy Plant, a plant to fight moths and avoid damages in crops.</p><br/><br />
<br />
<br />
<div align="center"><img width="400px" src="https://static.igem.org/mediawiki/2014/a/ac/VUPVOverview_figure_2.png" alt="circuit"></img></div><br/><br />
<br />
<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 2. Our system in an electric circuit-like format</span>. The plant’s own metabolism is used as the energy source. The biosafety module, composed by identity preservation and male sterility parts is always under expression in the system, to ensure the security of the transgenic plant. Pheromone synthetic pathways (A,B or C, depending on the problem pest) are activated using a genetic switch sensitive to Copper Sulfate, so system will not be producing pheromones unless it is activated with Copper-rich solution.</p></div><br/><br/><br/><br/><br/><br/><br />
<br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV"><strong>&larr; Go to Home</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules"><strong>Go to Modules</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/results"><strong>Go to Results &rarr;</strong></a></div><br/><br/><br/><br/><br />
<br />
<p align="center" class="black-bold">REFERENCES</p><br/><br />
<div style="position: relative; left: 3%; width: 96%;"><ol><br />
<li>Ding BJ, Hofvander P, Wang HL, Durrett TP, Stymne S, et al. (2014) A plant factory for moth pheromone production. Nat Commun 5: 3353.</li><br />
<li>Hagström A, Wang HL, Lienard MA, Lassance JM, Johansson T, et al. (2013) A moth pheromone brewery: production of (Z)-11-hexadecenol by heterologous co-expression of two biosynthetic genes from a noctuid moth in a yeast cell factory. Microb Cell Fact 12: 125.</li><br />
</ol></div><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesisTeam:Valencia UPV/Project/modules/biosynthesis2014-10-17T22:39:48Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/modules">Modules</a> > <a>Pheromone Biosynthesis</a> </h3></p><br/></br><br />
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<div align="center"><span class="coda"><roja>P</roja>heromone <roja>B</roja>iosynthesis</span> </div><br/><br/><br />
<br />
<p><strong>Biosynthesis</strong></p><br/><br />
<br />
<p>This is the core of our project. <span class="red-bold">Sexy Plant</span> is a pest control choice based on the production of sexual pheromones, which will confuse male insects and make them unable to find the female. For that reason, engineering the genome of <span class="italic">Nicotiana benthamiana</span> to produce a set of sexual pheromones was our priority.</p><br/><br/><br />
<br />
<p>We wanted to produce pheromones to generate mating disruption in species which cause great damage in crops, so we did some research and consulted experts in the CEQA . We also took into account that they should be species with already identified sexual pheromones as well as their biosynthetic pathways. We found the perfect group of species to fight against, Moths.</p><br/><br/><br />
<br />
<p>Sexual pheromones for a large number of moths have already been identified. Most of these species use type I pheromones. These consist of “straight-chain compounds 10-18 carbons in length with a functional group of a primary alcohol, aldehyde, or acetate ester, and usually with several double bonds” [1]. Moths de novo synthesize these pheromones in the pheromone gland (PG) through modifications of fatty acid biosynthetic pathways” [5] Three of the most commonly found major components compounds are <span class="blue-bold">Z11-16:OAc</span>, <span class="green-bold">Z11-16:Ald</span> and <span class="red-bold">Z11-16:OH</span> [1] (see figure 1). In addition, biosynthetic pathways of these pheromones had already been elucidated in some species [2-5], which was a great help to create our biosynthetic pathways.</p><br/><br/><br/><br />
<br />
<br />
<div align="center"><img width="320px" src="https://static.igem.org/mediawiki/2014/8/80/VUPVBiosynthesis_figure_1.png" alt="pheromone_structures" title="Moths Pheromones Structure"></img></div><br/><br />
<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1. Structure of moths sexual pheromones</span>. A: Structure of <span class="red-bold">Z11-16:OH</span>. B:Structure of <span class="green-bold">Z11-16:Ald</span>. C: Structure of <span class="blue-bold">Z11-16:OAc</span>.</p></div><br/><br/><br />
<br />
<br />
<p>In previous transcriptome analysis from the moth <span class="italic">Agrotis ipsilon</span>, specific sets of enzymes were identified to be differentially expressed in the pheromone glands. These enzymes were Acetyl-CoA carboxylases, fatty acid synthases, desaturases, acyl-CoA reductases, alcohol oxidases, aldehyde reductases and acetyltransferases. They were found to be differentially expressed in the moths pheromone glands compared to the rest of the organism [5]. These results mean these enzymes are involved in pheromones biosynthesis.</p><br/><br/><br />
<br />
<p><strong>Previous approaches</strong></p><br/><br />
<br />
<p>Moth sexual pheromones had already been synthesized previously using different approaches: Hagström et al produced <span class="red-bold">Z11-16:OH</span> and <span class="green-bold">Z11-16:Ald</span> in yeast [6]. Ding et al produced <span class="blue-bold">Z11-16:OAc</span> in <span class="italic">Nicotiana benthamiana</span>, our chassis plant, by transient expression of individual enzymes [7]. However, the production of these three pheromones in plant had never been tested using an in cis multigenic construction … until we arrived.</p><br/><br/><br />
<br />
<p><strong>Our system</strong></p><br/><br />
<br />
<p>We created a plant able to synthesize sexual pheromones <span class="blue-bold">Z11-16:OAc</span>, <span class="green-bold">Z11-16:Ald</span> and <span class="red-bold">Z11-16:OH</span>, the most important ones in moths sexual behavior (see the tables 1 to 3 below). Sexy Plant expresses sets of genes which transform Palmitic Acid CoA, an abundant compound in <span class="italic">N.benthamiana</span> leafs, into these three pheromones. Depending on the pheromone to be produced, different pathways are introduced in Biobricks standard and expressed in the plant.</p><br/><br/><br />
<br />
<img width="250px" style="float:right;" src="https://static.igem.org/mediawiki/2014/3/3c/VUPVBiosynthesis_side.png" alt="plants_moths"></img><br/><br />
<br />
<p><br/>As it was done in Ding's work [7], a Δ11 desaturase from <span class="italic">Amyelois transitella</span> (accession number JX964774) and HarFAR_KKYR, an improved version of HarFAR-3 fatty acid reductase from <span class="italic">Helicoverpa armigera</span> (accession number JF709978) were expressed to produce <span class="red-bold">Z11-16:OH</span>. Additionally, 1,2-diacyl-sn-glycerol:acetyl-CoA acetyltransferase from <span class="italic">Euonymus alatus</span> (accession number GU594061) was expressed along with the other two enzymes to produce <span class="blue-bold">Z11-16:OAc</span> [7]. Inspired by Hagström approach [6], production of <span class="green-bold">Z11-16:Ald</span> could be accomplished by over expressing a fatty-acid alcohol oxidase from <span class="italic">Nicotiana benthamiana</span>. The whole biosynthetic pathway is depicted in figure 2.</p><br/><br/><br/><br/><br />
<br />
<br />
<div align="center"><img width="450px" src="https://static.igem.org/mediawiki/2014/f/fd/VUPVBiosynthesis_figure_2.png" alt="pheromone_pathway" title="Pheromones Pathway"></img></div><br/><br />
<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 2. Biosynthetic pathway of moth sexual pheromones</span>. Sexual pheromones are bordered in purple. Taking Palmitic Acid CoA (16:CoA) as substrate, the expression of the genes AtrΔ11 (Desaturase) and HarFAR (Reductase) leads to the production of <span class="red-bold">Z11-16:OH</span>. By adding the gene EaDAcT (Acetyltransferase) in the previous system, production displaced to obtaining <span class="blue-bold">Z11-16:OAc</span>. If a Alcohol Oxidase (FAO) is included instead of EaDAcT, <span class="green-bold">Z11-16:Ald</span> will be produced.</p></div><br/><br/><br />
<br />
<br />
<p><strong>Finding pests</strong></p><br/><br />
<br />
<p>Complementing our efforts to produce the pheromones, we did a thorough search in the Pherobase database (http://www.pherobase.com) with software we specifically developed for this purpose and checked which insects met the conditions both to have one of the pheromones produced by the Sexy Plant as a major pheromone component and also to be considered a plague. Results from this analysis can be found in the tables below. We can conclude that there is a large number of insects causing plagues that are affected by our pheromones.</p><br/><br/><br />
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<br/><br/><br/><br />
<br />
<p>Insects attracted by <span class="green-bold">Z11-16:Ald</span> and <span class="red-bold">Z11-16:OH</span></p><hr/><br/><br/><br />
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<div align="center"><table class="normal-table"><br />
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<tr><br />
<th>Importance</th><br />
<th>Insect</th><br />
<th>Plague</th><br />
</tr><br />
<br />
<tr><br />
<td>High</td><br />
<td>Helicoverpa armigera</td><br />
<td>Many crops, cotton, ornamentals, fruit trees...</td><br />
</tr><br />
<br />
<tr><br />
<td>High</td><br />
<td>Helicoverpa zea</td><br />
<td>Many crops, cotton, linum...</td><br />
</tr><br />
<br />
<tr><br />
<td>High</td><br />
<td>Heliothis peltigera</td><br />
<td>Cotton</td><br />
</tr><br />
<br />
<tr><br />
<td>High</td><br />
<td>Heliothis virescens</td><br />
<td>Many crops, cotton, tobacco, fruit trees...</td><br />
</tr><br />
<br />
<tr><br />
<td>Medium</td><br />
<td>Gortyna xanthenes</td><br />
<td>Artichoke (Comunidad Valenciana)</td><br />
</tr><br />
<br />
<tr><br />
<td>Medium</td><br />
<td>Platyptilia carduidactyla</td><br />
<td>Artichoke</td><br />
</tr><br />
<br />
<tr><br />
<td>Medium</td><br />
<td>Diatraea considerata</td><br />
<td>Sugar cane</td><br />
</tr><br />
<br />
</table></div><br/><br/><br/><br/><br />
<br />
<p>Insects attracted by <span class="blue-bold">Z11-16:OAc</span> and <span class="red-bold">Z11-16:OH</span></p><hr/><br/><br/><br />
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<div align="center"><table class="normal-table"><br />
<br />
<tr><br />
<th>Importance</th><br />
<th>Insect</th><br />
<th>Plague</th><br />
</tr><br />
<br />
<tr><br />
<td>Medium</td><br />
<td>Crocidolomia binotalis</td><br />
<td>Cabbage</td><br />
</tr><br />
<br />
<tr><br />
<td>Medium</td><br />
<td>Mamestra brassicae</td><br />
<td>Coliflower</td><br />
</tr><br />
<br />
<tr><br />
<td>Medium</td><br />
<td>Feltia jaculifera</td><br />
<td>Maize, sorghum</td><br />
</tr><br />
<br />
<tr><br />
<td>Medium</td><br />
<td>Euxoa messoria</td><br />
<td>Apple, cultivated vegetables, flowers...</td><br />
</tr><br />
<br />
<tr><br />
<td>Medium</td><br />
<td>Sesamia calamistis</td><br />
<td>Sugar cane</td><br />
</tr><br />
<br />
<tr><br />
<td>Medium</td><br />
<td>Platyptilia carduidactyla</td><br />
<td>Maize</td><br />
</tr><br />
<br />
</table></div><br/><br/><br/><br/><br />
<br />
<p>Insects attracted by <span class="red-bold">Z11-16:OH</span></p><hr/><br/><br/><br />
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<div align="center"><table class="normal-table"><br />
<br />
<tr><br />
<th>Importance</th><br />
<th>Insect</th><br />
<th>Plague</th><br />
</tr><br />
<br />
<tr><br />
<td>Medium</td><br />
<td>Chilo zacconius</td><br />
<td>Rice</td><br />
</tr><br />
<br />
</table></div><br/><br/><br/><br/><br />
<br />
<br />
<div align="center"><br />
<a class="button-content" id="goto-left" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/moths_behavior"><strong>&larr; Go to Moths Behaviour</strong></a><br />
<a class="button-content" id="goto-middle" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules"><strong>Go to Modules</strong></a><br />
<a class="button-content" id="goto-right" align="center" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/release"><strong>Go to Release &rarr;</strong></a></div></br></br></br><br/><br />
<br />
<p align="center"><strong>References</strong></p><br/><br />
<div style="position: relative; left: 3%; width: 96%;"><ol><br />
<li>Matsumoto S (2010) Molecular Mechanisms Underlying Sex Pheromone Production in Moths. Bioscience, Biotechnology, and Biochemistry 74: 223-231.</li><br />
<li>Choi M-Y, Han KS, Boo KS, Jurenka RA (2002) Pheromone biosynthetic pathways in the moths Helicoverpa zea and Helicoverpa assulta. Insect Biochemistry and Molecular Biology 32: 1353-1359.</li><br />
<li>Wang H-L, Zhao C-H, Wang C-Z (2005) Comparative study of sex pheromone composition and biosynthesis in Helicoverpa armigera, H. assulta and their hybrid. Insect Biochemistry and Molecular Biology 35: 575-583.</li><br />
<li>Fu X, Fukuzawa M, Tabata J, Tatsuki S, Ishikawa Y (2005) Sex pheromone biosynthesis in Ostrinia zaguliaevi, a congener of the European corn borer moth O. nubilalis. Insect Biochemistry and Molecular Biology 35: 621-626.</li><br />
<br />
<li>Gu S-H, Wu K-M, Guo Y-Y, Pickett J, Field L, et al. (2013) Identification of genes expressed in the sex pheromone gland of the black cutworm Agrotis ipsilon with putative roles in sex pheromone biosynthesis and transport. BMC Genomics 14: 636.</li><br />
<li>Hagström A, Wang H-L, Lienard M, Lassance J-M, Johansson T, et al. (2013) A moth pheromone brewery: production of (Z)-11-hexadecenol by heterologous co-expression of two biosynthetic genes from a noctuid moth in a yeast cell factory. Microbial Cell Factories 12: 125.</li> <br />
<li>Ding BJ, Hofvander P, Wang HL, Durrett TP, Stymne S, et al. (2014) A plant factory for moth pheromone production. Nat Commun 5: 3353.</li> <br />
</ol><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/VUPV_PartsTeam:Valencia UPV/VUPV Parts2014-10-17T22:33:45Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Achievements</a> > <a>Parts</a></h3></p><br/><br />
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<div align="center"><span class="coda"><roja>P</roja>arts</span> </div><br/><br />
<p>Our team sent 7 BioBrick Parts to the <a class="normal-link-page" href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2014&group=Valencia_UPV">Registry</a>.</p><br/><br />
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<div align="center"><groupparts>iGEM014 Valencia_UPV</groupparts></div><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554000">BBa_K1554000</a> – Ta29 Promoter</b></p><br />
<p>The Ta29 promoter is anther-specific. Anthers are the male organs of the flower, and their function is to produce pollen. A cytotoxic gene (barnase, ipt, RIP...) is usually used under the control of this specific promoter leading to a plant unable to produce pollen, inducing male-sterility. <a href=https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety> See Biosafety</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554001">BBa_K1554001</a> – AtrΔ11</b></p><br />
<p>The AtrΔ11 protein is a delta-11-desaturase from Amyelois transitella that introduces an unsaturation between C11 and C12 in long-chain fatty acids. This DNA sequence underwent codon usage optimization for Nicotiana benthamiana.<br />
Acyl-CoA + Reduced acceptor + O2 = Delta11-acyl-CoA + Acceptor + 2 H2O<br />
This part was used in the Insect sexual pheromone production pathway. <a href=https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesis> See Biosynthesis</p><br/><br />
<br />
<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554002">BBa_K1554002</a> – HarFAR</b></p><br />
<p>The HarFAR protein is a fatty acid reductase that catalyses the conversion of a long-chain fatty acid carboxyl group to an alcohol group. The DNA sequence was obtained from Helicoverpa armigera HarFAR-3 protein, including an Endoplasmic Reticulum retention signal (KKYR) in the C-terminal end. In addition, codon usage optimization was performed for Nicotiana benthamiana.<br />
This part was used in the Insect sexual pheromone production pathway. <a href=https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesis> See Biosynthesis</p><br/><br />
<br />
<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554003">BBa_K1554003</a> – EaDAcT</b></p><br />
<p>The EaDAcT protein is a diacylglycerol acetyltransferase coming from Euonymus alatus. This enzyme can transform alcohol gropus from fatty acid to aldehyde gropus. This DNA sequence underwent codon usage optimization for Nicotiana benthamiana.<br />
This part was used in the Insect sexual pheromone production pathway. <a href=https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosynthesis> See Biosynthesis </a></p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554004">BBa_K1554004</a> – Yellow biosafety module for plants</b></p><br />
<p>This biosafety device for plants consists of two submodules: a male sterility submodule and an identity preservation submodule. The male sterility submodule consists of a barnase specifically expressed in anthers under the regulation of the TA29 tapetum-specific promoter. As result, pollen from these plants is not fertile. The identity preservation submodule expresses the yellow chromoprotein AmilGFP, so that plants can be visually differentiated from non-transgenic plants. See Results: Constructs-Biosafety</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554005">BBa_K1554005</a> - Blue biosafety module for plants</b></p><br />
<p>This biosafety device is composed of the same parts as the previously explained one, BBa_K1554004. This device uses AmilCP as chromoprotein instead of AmilGFP. See Results: Constructs-Biosafety</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554006">BBa_K1554006</a> - Omega Undercover</b></p><br />
<p>The Omega undercover vector allows the conversión of GoldenBraid standard parts to BioBrick’s. <a href=https://2014.igem.org/Team:Valencia_UPV/Project/modules/methodology/parts_construction> See Methodology: Parts construction </a></p><br/><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/VUPV_PartsTeam:Valencia UPV/VUPV Parts2014-10-17T22:10:25Z<p>Alquiru: </p>
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<div>{{:Team:Valencia_UPV/header}}<br />
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<br />
<div align="center"><span class="coda"><roja>P</roja>arts</span> </div><br/><br />
<p>Our team sent 7 BioBrick Parts to the <a class="normal-link-page" href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2014&group=Valencia_UPV">Registry</a>.</p><br/><br />
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<br />
<div align="center"><groupparts>iGEM014 Valencia_UPV</groupparts></div><br />
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<html></br><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554000">BBa_K1554000</a> – Ta29 Promoter</b></p><br />
<p>The Ta29 promoter is anther-specific. Anthers are the male organs of the flower, and their function is to produce pollen. A cytotoxic gene (barnase, ipt, RIP...) is usually used under the control of this specific promoter leading to a plant unable to produce pollen, inducing male-sterility. (see Biosafety)</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554001">BBa_K1554001</a> – AtrΔ11</b></p><br />
<p>The AtrΔ11 protein is a delta-11-desaturase from Amyelois transitella that introduces an unsaturation between C11 and C12 in long-chain fatty acids. This DNA sequence underwent codon usage optimization for Nicotiana benthamiana.<br />
Acyl-CoA + Reduced acceptor + O2 = Delta11-acyl-CoA + Acceptor + 2 H2O<br />
This part was used in the Insect sexual pheromone production pathway (link Biosynthesis)</p><br/><br />
<br />
<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554002">BBa_K1554002</a> – HarFAR</b></p><br />
<p>The HarFAR protein is a fatty acid reductase that catalyses the conversion of a long-chain fatty acid carboxyl group to an alcohol group. The DNA sequence was obtained from Helicoverpa armigera HarFAR-3 protein, including an Endoplasmic Reticulum retention signal (KKYR) in the C-terminal end. In addition, codon usage optimization was performed for Nicotiana benthamiana.<br />
This part was used in the Insect sexual pheromone production pathway (link Biosynthesis)</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554003">BBa_K1554003</a> – EaDAcT</b></p><br />
<p>The EaDAcT protein is a diacylglycerol acetyltransferase coming from Euonymus alatus. This enzyme can transform alcohol gropus from fatty acid to aldehyde gropus. This DNA sequence underwent codon usage optimization for Nicotiana benthamiana.<br />
This part was used in the Insect sexual pheromone production pathway (link Biosynthesis)</p><br/><br />
<br />
<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554004">BBa_K1554004</a> – Yellow biosafety module for plants</b></p><br />
<p>This biosafety device for plants consists of two submodules: a male sterility submodule and an identity preservation submodule. The male sterility submodule consists of a barnase specifically expressed in anthers under the regulation of the TA29 tapetum-specific promoter. As result, pollen from these plants is not fertile. The identity preservation submodule expresses the yellow chromoprotein AmilGFP, so that plants can be visually differentiated from non-transgenic plants.</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554005">BBa_K1554005</a> - Blue biosafety module for plants</b></p><br />
<p>This biosafety device is composed of the same parts as the previously explained one, BBa_K1554004. This device uses AmilCP as chromoprotein instead of AmilGFP.</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554006">BBa_K1554006</a> - Omega Undercover</b></p><br />
<p>The Omega undercover vector allows the conversión of GoldenBraid standard parts to BioBrick’s. Methodology: Parts contruction</p><br/><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/VUPV_PartsTeam:Valencia UPV/VUPV Parts2014-10-17T22:07:58Z<p>Alquiru: </p>
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<br />
<div align="center"><span class="coda"><roja>P</roja>arts</span> </div><br/><br />
<p>Our team sent 7 BioBrick Parts to the <a class="normal-link-page" href="http://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2014&group=Valencia_UPV">Registry</a>.</p><br/><br />
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<div align="center"><groupparts>iGEM014 Valencia_UPV</groupparts></div><br />
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<html></br><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554000">BBa_K1554000</a> – Ta29 Promoter</b></p><br />
<p>The Ta29 promoter is anther-specific. Anthers are the male organs of the flower, and their function is to produce pollen. A cytotoxic gene (barnase, ipt, RIP...) is usually used under the control of this specific promoter leading to a plant unable to produce pollen, inducing male-sterility. (see Biosafety)</p><br/><br />
<br />
<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554001">BBa_K1554001</a> – AtrΔ11</b></p><br />
<p>The AtrΔ11 protein is a delta-11-desaturase from Amyelois transitella that introduces an unsaturation between C11 and C12 in long-chain fatty acids. This DNA sequence underwent codon usage optimization for Nicotiana benthamiana.<br />
Acyl-CoA + Reduced acceptor + O2 = Delta11-acyl-CoA + Acceptor + 2 H2O<br />
This part was used in the Insect sexual pheromone production pathway (link Biosynthesis)</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554002">BBa_K1554002</a> – HarFAR</b></p><br />
<p>The HarFAR protein is a fatty acid reductase that catalyses the conversion of a long-chain fatty acid carboxyl group to an alcohol group. The DNA sequence was obtained from Helicoverpa armigera HarFAR-3 protein, including an Endoplasmic Reticulum retention signal (KKYR) in the C-terminal end. In addition, codon usage optimization was performed for Nicotiana benthamiana.<br />
This part was used in the Insect sexual pheromone production pathway (link Biosynthesis)</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554003">BBa_K1554003</a> – EaDAcT</b></p><br />
<p>The EaDAcT protein is a diacylglycerol acetyltransferase coming from Euonymus alatus. This enzyme can transform alcohol gropus from fatty acid to aldehyde gropus. This DNA sequence underwent codon usage optimization for Nicotiana benthamiana.<br />
This part was used in the Insect sexual pheromone production pathway (link Biosynthesis)</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554004">BBa_K1554004</a> – Yellow biosafety module for plants</b></p><br />
<p>This biosafety device for plants consists of two submodules: a male sterility submodule and an identity preservation submodule. The male sterility submodule consists of a barnase specifically expressed in anthers under the regulation of the TA29 tapetum-specific promoter. As result, pollen from these plants is not fertile. The identity preservation submodule expresses the yellow chromoprotein AmilGFP, so that plants can be visually differentiated from non-transgenic plants.</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554005">BBa_K1554005</a> - Blue biosafety module for plants</b></p><br />
<p>This biosafety device is composed of the same parts as the previously explained one, BBa_K1554004. This device uses AmilCP as chromoprotein instead of AmilGFP.</p><br/><br />
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<p> <b><a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1554006">BBa_K1554006</a> - Omega Undercover</b></p><br />
<p>The Omega undercover vector allows the conversión of GoldenBraid standard parts to BioBrick’s. Methodology: Parts contruction</p><br/><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Team/Students/Lucia_EstellesTeam:Valencia UPV/Team/Students/Lucia Estelles2014-10-17T21:47:16Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Team</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students">Students</a> > <a>Lucía Estellés López</a></h3></p><br />
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<img alt="Profile_Jana" title="Lucía Estellés" src="https://static.igem.org/mediawiki/2014/a/af/VUPVProfile_Lucia.jpg" class="img-profile-desc"></img><br />
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<h4 class="title-desc">Lucía Estellés</h4><br/><br />
<p>Lucía is currently undertaking a double degree in Biotechnology BSc at the Polytechnic University of Valencia and Applied Bioinformatics MSc at Cranfield University. She loved participating in the iGEM competition as she had a great opportunity to learn more about Systems Biology.</p><br/><br />
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Lucía likes learning new things, the most diverse as possible, and reading. </p><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Team/Students/Lucia_EstellesTeam:Valencia UPV/Team/Students/Lucia Estelles2014-10-17T21:44:53Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Team</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students">Students</a> > <a>Lucía Estellés López</a></h3></p><br />
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<h4 class="title-desc">Lucía Estellés</h4><br/><br />
<p>Lucía is currently undertaking a double degree in Biotechnology BSc at the Polytechnic University of Valencia and Applied Bioinformatics MSc at Cranfield University. She loved participating in the iGEM competition as she had a great opportunity to learn more about Systems Biology.</p><br/><br />
<p><br />
Lucía likes learning new things, the most diverse as possible, and reading. </p><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Team/StudentsTeam:Valencia UPV/Team/Students2014-10-17T21:41:17Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Team</a> > <a>Students</a></h3></p></br><br />
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<div align="center"><span class="coda"><roja>S</roja>tudents</span> </div><br />
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<p><h4>Alba Rubert</h4><br />
Biotechnology<br/><br/><br/><br/><br/><br/>For more details: click <a href="#">here</a><br />
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<p><h4>Alejandra González</h4><br />
Engineering<br/><br/><br/><br/><br/><br/>For more details: click <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students/Alejandra_Gonzalez">here</a><br />
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<p><h4>Alfredo Quijano</h4><br />
Biotechnology<br/><br/><br/><br/><br/><br/>For more details: click <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students/Alfredo_Quijano">here</a><br />
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<img alt="Profile_Ivan" class="img-profile" title="Ivan Llopis" src="https://static.igem.org/mediawiki/2014/4/4b/Valencia_UPV_Profile_Ivan_mod.png"></img><br />
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<p><h4>Ivan Llopis</h4><br />
Engineering<br/><br/><br/><br/><br/><br/>For more details: click <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students/Ivan_Llopis">here</a><br />
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<p><h4>Jose Gavaldá</h4><br />
Biotechnology<br/><br/><br/><br/><br/><br/>For more details: click <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students/Jose_Gavalda">here</a><br />
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<p><h4>Lucía Estellés</h4><br />
Biotechnology<br/><br/><br/><br/><br/><br/>For more details: click <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students/Lucia_Estelles">here</a><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Project/results/biosafetyTeam:Valencia UPV/Project/results/biosafety2014-10-17T21:37:24Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Project</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Project/results">Results</a> > <a>Biosafety</a></h3></p><br/><br/><br />
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<div align="center"><span class="coda"><roja>B</roja>iosafety</span> </div><br/><br/><br />
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<p>Our goal was to develop a sterile and easily identifiable plant. In order to do this, we created a module in collaboration with <a href="https://2014.igem.org/Team:NRP-UEA-Norwich" class="normal-link-page">NRP-UEA-Norwich</a> which incorporated an RNAse (barnase) under the regulation of a tapetum specific promoter (TA29) and a chromoprotein</p><br/><br />
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<p>Male Sterility strategy was not possible to test in a transient approach, but both components are well documented (<a class="normal-link-page" href="https://2014.igem.org/Team:Valencia_UPV/Project/modules/biosafety">see Biosafety Module</a>).</p><br/><br />
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<p>The NRP-UEA-Norwich provided us with blue and yellow chromoproteins transcriptional units (TU) and we agroinfiltrated both TUs in <i>N. benthamiana</i> along with a GFP control. Chromoprotein detection was impossible by the naked eye (Figure 1) even though GFP control was expressed (Figure 2). Plants agroinfiltration was correct since GFP was expressed in the leaf.</p><br/><br/><br />
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<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 1. Plants agroinfiltrated with NRP-UEA-Norwich chromoproteins</span>. TUs containing Blue (left) and Yellow (Right) chromoproteins were agroinfiltrated in N.benthamiana. None of the chromoproteins could be detected by the naked eye.</p></div><br/><br/><br />
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<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 2. Agroinfiltration GFP control</span>. Agroinfiltration control shows GFP expression, indicating that non-detection of chromoproteins is not due to a failure in agroinfiltration.</p></div><br/><br/><br />
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<p>Nevertheless, identity preservation is an important part of the biosafety module, and it must be happen efficiently. <a href="https://2014.igem.org/Team:NRP-UEA-Norwich" class="normal-link-page">NRP-UEA-Norwich</a> team suggested leaf degreening in order to observe the chromoproteins, but we don’t consider this strategy convenient for our purpose since identity should be easily recognisable in plants without any kind of treatment. As an alternative to chromoproteins, we propose the use of two transcriptional factors to enhance anthocyanins production.</p><br/><br />
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<p>Our identity preservation construct consists of two transcriptional units carrying the tomato (<i>Solanum lycopersicum</i>) transcriptional factors (SlANT1, SlJAF13) which are regulated by the 35S constitutive promoter. Both transcriptional factors are involved in flavonoids biosynthetic pathway regulation; it is that they enhance anthocyanin accumulation.</p><br/><br />
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<p>SlANT1 and SlJAF13 are <i>S. lycopersicum</i> orthologous of the <i>Antirrhinum majus</i> Rosea1 and Delila genes. In previous researchs, ectopic over-expression of these transcription factors under the control of the E8 fruit-specific promoter increases the transcript levels of most of the anthocyanin biosynthetic genes in tomato fruit leading to high levels of <i>anthocyanins</i> [1,2]. In <i>N. benthamiana</i>, transient expression of SIANT1 and SIJAF3 activates the expression of several flavonoid biosynthetic genes leading to a change of the colour on the leaf due to accumulation of anthocyanins.</p><br/><br />
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<p>We transiently transformed this new identity preservation module (containing SIANT1 and SIJAF13 transcriptional units) by agroinfiltration into <i>N. benthamiana</i>. As result, the anthocyanin accumulation drives our plant to a violet colour change that can be observed by the naked eye (Figure 3).</p><br/><br />
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<div align="center"><p style="text-align: justify; font-style: italic; font-size: 0.8em; width: 700px;"><span class="black-bold">Figure 3. Anthocyanins accumulation in <i>N.benthamiana</i></span>. SlANT1 and SlJAF13 expression produces accumulation of anthocyanins in agroinfiltrated plant (Left), showing a purple colour in contrast with a green wild type leaf (Right, held by hand).</p></div><br/><br/><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/File:VUPV_Rosea.pngFile:VUPV Rosea.png2014-10-17T21:34:55Z<p>Alquiru: </p>
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<div></div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/policy/activitiesTeam:Valencia UPV/policy/activities2014-10-17T20:55:12Z<p>Alquiru: </p>
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<div align="center"><span class="coda"><roja>A</roja>ctivities</span> </div><br/><br/><br />
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<p>This are the activities and meetings in which we participated:</p><br />
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<h3><b>Permanent activities and meetings</b></h3><br />
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<h3> Center for Chemical Ecology (CEQA), Valencia.</h3><br />
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CEQA has expertise in the isolation and identification of semiochemicals, the formulation of pheromone attractants and controlled rate emitters.</p><br />
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Jaime Primo Millo is the Director of the Agricultural Center for Chemical Ecology (CEQA), and professor of organic chemistry at UPV. Jaime Primo, Ismael Navarro, Vicente Navarro and Sandra Vacas have continuously advised the Valencia UPV iGEM team since we first met them. They are developing systems to control the principal citrus pests, and their experience on “insects sexual confusion” has been invaluable for us, and has improved the achievements of our <span class="red-bold">Sexy Plant</span> as a new pest management method. <br />
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<h3><i>"Projects like The Sexy Plant show us the importance Synthetic Biology may have in the future of agriculture"</i></h3></html><br />
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<p>Mr. Jorge Silva is the Head of Bayer CropScience Technical Department. Jorge liked <span class="red-bold">Sexy Plant</span> as a system to fight against pests. He highlighted the necessity to find new approaches in sustainable agriculture, and that our project could have future in the market. We discussed the advances in plant synthetic biology and he concluded “projects like Sexy Plant show us the importance Synthetic Biology may have in the future of agriculture”. Jorge gave us invaluable feedback on how to improve certain details to ease the commercialization of Sexy Plant in the future, and asked us to send an executive summary of our project to Bayer CropScience headquarters in Ghent. As result of the meeting, Bayer CropScience wrote an official support letter for our project, and asked to be informed about further developments of the project in the future. </p><br />
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<h3><i>“Sexy Plant could be a good approach to reduce the costs of pheromone production” </i></h3><br />
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<p>Jose Maria Garcia Alvarez-Coque is a social researcher and Director of the Sustainable Agriculture group, at the Universitat Politècnica de Valencia (UPV). </p><br />
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He considers the currently pheromone synthesis as an expensive method to produce insect pheromones. “Sexy Plant could be a good approach to reduce the costs of pheromone production”. This project is another ecological way to manage pests, such as the production of auxiliary insects to protect crops. In addition, Sexy Plant can be a safe, sustainable and environmentlly friendly method, as far as it respects agricultural and environmental specifications. <span class="red-bold">Sexy Plant</span> modules like Identity Preservation or Sterility are characteristics that assist the long-term use of this plant. </p><br/><br />
<p align="right"> Valencia, September 2014</p><br />
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<h3><i>“We have been using sexual confusion as a pest management method in rice crops for the last 20 years” </i></h3><br />
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Paco Girona is agriculture engineer at Cooperatives for Agrofood industry (FECOAV). He makes efforts to extend the use of ecological pest methods over some typical crops in Valencia, Spain. </p><br />
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Around the Albufera Natural Park in Valencia, the cycle sown rice starts in May when the farmers should start protecting their crops from the <i>Chilo</i> lepidopter (moth) pest. During the first days of May, technicians place pheromone sticks between rice plants to release the Chilo female moth pheromone into the environment. The planted area is around 15,000 Ha of crop, and they are protected by 462,960 pheromone sticks. This technique has been used for the last 20 years, and the economic losses have been cut down to approximately a mere 0.5%. Paco Girona explains that Sexy Plant coul be a wonderful alternative to chemically synthetized pheromone sticks, specially during its assembly, the most hazardous part of its production. “If you get the <span class="red-bold">Sexy Plant</span>, I take my hat off” </p><br />
<p align="right"> Valencia, October 2014</p><br />
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<p>Our team also talked with Professor Sam Tothill, Professor of Biosensors in Health at Cranfield University, United Kingdom. Her current research mainly focuses in biosensors for microbial contaminants and pathogen's (bacteria, viruses, fungi) and their toxins. We really appreciated her feedback as part of her research consist in the detection of contaminants such as pesticides in food. We discussed about safety issues and how could our project be implemented in agriculture. She really liked the about the <span class="red-bold">Sexy Plant</span> project and wished our team succeeded at iGEM. </p><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Team/Students/Lucia_EstellesTeam:Valencia UPV/Team/Students/Lucia Estelles2014-10-17T20:51:57Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Team</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students">Students</a> > <a>Lucía Estellés López</a></h3></p><br />
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<h4 class="title-desc">Lucía Estellés</h4><br/><br />
<p>Lucía is currently undertaking a double degree in Biotechnology BSc at the Polytechnic University of Valencia and Applied Bioinformatics MSc at Cranfield University. She loved participating in the iGEM competition as she had a great opportunity to learn more about Systems Biology.</p><br/><br />
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Lucía likes learning new things, the most diverse as possible, and reading. </p><br />
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{{:Team:Valencia_UPV/footer_img}}</div>Alquiruhttp://2014.igem.org/Team:Valencia_UPV/Team/Students/Lucia_EstellesTeam:Valencia UPV/Team/Students/Lucia Estelles2014-10-17T20:51:19Z<p>Alquiru: </p>
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<p><h3 class="hook" align="left"><a>Team</a> > <a href="https://2014.igem.org/Team:Valencia_UPV/Team/Students">Students</a> > <a>Lucía Estellés López</a></h3></p><br />
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<h4 class="title-desc">Lucía Estellés</h4><br/><br />
<p>Lucía is currently undertaking a double degree in Biotechnology BSc at the Polytechnic University of Valencia and Applied Bioinformatics MSc at Cranfield University. She loved participating in the iGEM competition as she had a great oportunity to learn more about Systems Biology.</p><br/><br />
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Lucía likes learning new things, the most diverse as possible, and reading. </p><br />
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