Difference between revisions of "Team:Washington"

 
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        <ul>
                     <p><a href="https://2015.igem.org/Team:Example2/Practices">HUMAN PRACTICES</a></p>
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            <a href="https://2015.igem.org/Team:Washington">
                     <p><a href="https://2015.igem.org/Team:Example2/Safety">SAFETY</a></p>
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                <li>Home
                     <p><a href="https://2015.igem.org/Team:Example2/Modeling">MODELING</a></p>
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                  <ul>
                     <p><a href="https://2015.igem.org/Team:Example2/Measurement">MEASUREMENT</a></p>
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                     <li><a href="https://2014.igem.org/Team:Washington">UW 2014</a></li>
                     <p><a href="https://2015.igem.org/Team:Example2/Software">SOFTWARE</a></p>
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                    <li><a href="https://2013.igem.org/Team:Washington">UW 2013</a></li>
                     <p><a href="https://2015.igem.org/Team:Example2/Entrepreneurship">ENTREPRENEURSHIP</a></p>
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                    <li><a href="https://2012.igem.org/Team:Washington">UW 2012</a></li>
                 </div>
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                    <li><a href="https://2011.igem.org/Team:Washington">UW 2011</a></li>
 +
                    <li><a href="https://2010.igem.org/Team:Washington">UW 2010</a></li>
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                    <li><a href="https://2009.igem.org/Team:Washington">UW 2009</a></li>
 +
                    <li><a href="https://2008.igem.org/Team:University_of_Washington">UW 2008</a></li>
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                    <li><a href="https://2013.igem.org/Main_Page">iGEM Homepage</a></li>
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                  </ul>
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</li>
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            </a>
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            <a href="https://2015.igem.org/Team:Washington/Auxin">
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                 <li>Auxin
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                    <div class="collapse navbar-collapse" id="bs-example-navbar-collapse-1">
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                        <ul class="nav navbar-nav">
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                            <li>
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                                <a href="https://2015.igem.org/Team:Washington/Auxin#Introduction">Introduction</a>
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                            </li>
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                            <li>
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                                <a href="https://2015.igem.org/Team:Washington/Auxin#Methods">Methods</a>
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                            </li>
 +
                            <li>
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                                <a href="https://2015.igem.org/Team:Washington/Auxin#Results">Results</a>
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                            </li>
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                            <li>
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                                <a href="https://2015.igem.org/Team:Washington/Auxin#Conclusion">Conclusion</a>
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                            </li>
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                            <li>
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                                <a href="https://2015.igem.org/Team:Washington/Parts#Auxin">Biobrick</a>
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                            </li>
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                        </ul>
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                     </div>
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                </li>
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            </a>
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            <a href="https://2015.igem.org/Team:Washington/Aptazyme">
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                <li>Aptazyme
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                    <ul>
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                            <li>
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                                <a href="https://2015.igem.org/Team:Washington/Aptazyme#Introduction">Introduction</a>
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                            </li>
 +
                            <li>
 +
                                <a href="https://2015.igem.org/Team:Washington/Aptazyme#Methods">Methods</a>
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                            </li>
 +
                            <li>
 +
                                <a href="https://2015.igem.org/Team:Washington/Aptazyme#Results">Results</a>
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                            </li>
 +
                            <li>
 +
                                <a href="https://2015.igem.org/Team:Washington/Aptazyme#Conclusion">Conclusion</a>
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                            </li>
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                            <li>
 +
                                <a href="https://2015.igem.org/Team:Washington/Parts#Aptazyme">Biobrick</a>
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                            </li>
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                     </ul>
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                </li>
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            </a>
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            <a href="https://2015.igem.org/Team:Washington/Paper_Device">
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                <li>Paper Device             
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                    <ul>
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                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Paper_Device#Introduction">Introduction</a>
 +
                        </li>
 +
                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Paper_Device#Methods">Methods</a>
 +
                        </li>
 +
                            <li>
 +
                                <a href="https://2015.igem.org/Team:Washington/Design">Design</a>
 +
                            </li>
 +
                            <li>
 +
                                <a href="https://2015.igem.org/Team:Washington/Paper_Device#Results">Results</a>
 +
                            </li>
 +
                            <li>
 +
                                <a href="https://2015.igem.org/Team:Washington/Paper_Device#Conclusion">Conclusion</a>
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                            </li>
 +
                     </ul>
 +
                </li>
 +
            </a>
 +
            <a href="https://2015.igem.org/Team:Washington/Modeling">
 +
                <li>Modeling             
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                    <ul>
 +
                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Modeling#Paper_Device">Paper Device</a>
 +
                        </li>
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                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Modeling#Aptazyme">Aptazyme</a>
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                        </li>
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                     </ul>
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                </li>
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            </a>
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            <a href="https://2015.igem.org/Team:Washington/Practices">
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                <li>Human Practices             
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                    <ul>
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                        <li>
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                            <a href="https://2015.igem.org/Team:Washington/Practices#Outreach">Outreach</a>
 +
                        </li>
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                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Practices#Integrated">Integrated</a>
 +
                        </li>
 +
                     </ul>
 +
                </li>
 +
            </a>
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            <a href="https://2015.igem.org/Team:Washington/Protocols">
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                <li>Protocols
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                    <ul>
 +
                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Protocols#Experiments">Experiments</a>
 +
                        </li>
 +
                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Protocols#Safety">Safety</a>
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                        </li>
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                     </ul>
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                </li>
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            </a>
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            <a href="https://2015.igem.org/Team:Washington/Team">
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                <li>Team                       
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                    <ul>
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                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Team#Members">Members</a>
 +
                        </li>
 +
                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Attributions">Attributions</a>
 +
                        </li>
 +
                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Team#Sponsors">Sponsors</a>
 +
                        </li>
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                        <li>
 +
                            <a href="https://2015.igem.org/Team:Washington/Team#Judging_Form">Judging Form</a>
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                        </li>
 +
                    </ul>
 +
                 </li>
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            </a>
 +
        </ul>
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    </div>
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    <!-- End of menu  -->
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    <!-- Start of content -->
  
<h2> Welcome to iGEM 2015! </h2>
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<br></br>
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        <h2 align=center> Lab on a Strip: Developing a Novel Platform for Yeast Biosensors </h2>
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        <h2> Project Overview </h2>
  
<h2> Project Description </h2>
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        <p>The Pacific Ocean is home to a wide range of marine life, including the food source of many filter-feeders, toxin-producing algae. When algal blooms are ingested by shellfish, the toxins produced by the algae are caught within shellfish tissue. Although these toxins are harmful to us, they aren’t to the shellfish, giving collectors no immediate sign of danger. Biotoxins are also just generally difficult to detect; contrary to popular belief, algal blooms are not always the striking crimson of “red tides.” Thus, blooms may not be discovered until after a poisoned shellfish is found. The Washington State Department of Health and commercial shellfish farmers conduct periodic surveys of local beaches to catch contaminations early, but these methods are costly, time-consuming, and not always effective. This can especially pose a dilemma for individual shellfish hunters, who do not have the resources to screen their shellfish for toxins.  With current detection methods, the crowds swarming to Seattle’s famous Pike Place Market and popular raw oyster bars are constantly at risk.</p><p><img src="https://static.igem.org/mediawiki/2015/f/ff/Igem_red_tide.jpeg" width=572 height=500 align="center" ></p>
  
<h2>Overview </h2>
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<br></br>
<p>For commercial shellfish farmers and recreational hunters alike, marine biotoxins pose a significant threat to health and welfare. With this project, we aim to create an inexpensive and easy-to-use test kit for the detection of the shellfish toxin okadaic acid using engineered yeast strains and DNA aptamers on a paper device. We also hope that this project paves the way for a new class of biosensors capable of detecting a wide range of small molecules. </p>
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        <p>We have developed a much cheaper diagnostic tool in which genetically-modified baker’s yeast is grown on a paper device and is able to produce an easy-to-read color output in the presence of a target molecule. Imagine if you could simply dip a sheet of paper into your bucket of shellfish, wait only (insert amount of time) and tell if your products are safe to consume. The proof-of-concept systems we’ve engineered detect the plant hormone auxin and the molecule theophylline. However, we’ve implemented a number of techniques to ensure the versatility of our systems thus, they can be easily modified and further developed to test for a wide variety of other molecules.</p>
  
<h2>What is the context of this research? </h2>
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  <body onload="init()">
<p>Marine toxins are an increasing problem in Washington State waters. Produced in high concentrations by microorganisms during algae blooms, they are ingested by filter-feeding shellfish, causing illness and death in human consumers. Biotoxins are also difficult to detect; contrary to popular belief, algal blooms are not always the striking crimson of “red tides.” Thus, blooms may not be discovered until after a poisoned shellfish is found. The Washington State Department of Health and commercial shellfish farmers conduct periodic surveys of local beaches to catch contaminations early, but these methods are costly, time-consuming, and not always effective. This can especially pose a dilemma for individual shellfish hunters, who do not have the resources to screen their shellfish for toxins. </p>
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    <ul id="tabs">
 +
      <li><a href="#Auxin">Auxin Pathway</a></li>
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      <li><a href="#Theophylline">Theophylline Pathway</a></li>
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    </ul>
  
<h2>What is the significance of this project? </h2>
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<div class="tabContent" id="Auxin">
<p>Our project aims to combine the emerging fields of synthetic biology and paper diagnostics to create an affordable, accessible, and accurate diagnostic test kit that would allow farmers and the public to test shellfish for common biotoxins. This “lab on a strip” will be a critical step forward in marine toxin detection, as it will cut nearly 20 hours off the time needed to obtain results, allowing farmers to screen at a lower cost and empowering individual hunters to confirm the safety of their shellfish. This is also the first project to attempt to grow yeast on a paper device, and if successful, could open the door to a wide range of similar biosensors. Such sensors would have applications in medicine, food, and the environment worldwide. </p>
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      <div>
 +
<p1>In the Auxin detection pathway, a DNA binding domain, a degron domain and a repressor domain are fused to suppress the expression of a reporter gene, LacZ. In the presence of Auxin, a plant hormone, along with a corresponding F-Box protein will lead to the fusion protein suppressing the reporter will be ubiquitinated allowing the reporter to be expressed.  </p1>
 +
      </div>
 +
    </div>
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<div class="tabContent" id="Theophylline">
 +
      <div>
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<p1>In this design RNA aptamers are used to sense our target molecule, theophylline.  Aptamers, unlike antibodies, can actually bind to virtually any molecule, allowing for a more versatile system.  We’ve implemented a ribozyme switch which, when active, cleaves the mRNA code of our target sequence, hindering the production of GFP by default. However, in the presence of theophylline our switch becomes inactive, allowing for the expression of our target gene.  This system is useful because it is faster-acting than more traditional expression pathways, and can be generalized to many other small molecules by changing the aptamer sequence.</p1>
 +
      </div>
 +
    </div>
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</body>
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<p><br><br></br></br></p>
  
<h2>What are the goals of the project?
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                                </div>
<p>1. Develop a paper microfluidic device that houses yeast; it will provide adequate nutrients for cell growth, but will also be freeze-dried for long-term storage.
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                            </div>
<p>2. Create a detection system for the plant hormone auxin in which yeast produce a color in response to an auxin input; test it in cells growing on normal media.
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<p>3. Clone each part from this system into standard plasmids for submission to BioBrick registry.
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<p>4. Show that when grown on paper, yeast can reliably detect auxin.
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<p>5. Use aptamers (short DNA sequences) to detect for okadaic acid, a shellfish toxin that causes Diarrhetic Shellfish Poisoning.
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<p>6. Show that the aptamer system for detection can be implemented on our paper platform and reliably detect for okadaic acid.
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<p>7. Improve safety of shellfish consumers in the NW and the world!
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<p>Your team has been approved and you are ready to start the iGEM season! </p>
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<h4>Before you start: </h4>
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<p> Please read the following pages:</p>
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<ul>
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<li>  <a href="https://2015.igem.org/Requirements">Requirements page </a> </li>
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<li> <a href="https://2015.igem.org/Wiki_How-To">Wiki Requirements page</a></li>
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</ul>
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<div class="highlightBox">
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<h4> Styling your wiki </h4>
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<p>You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.</p>
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<p>While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.</p>  
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<h4> Editing your wiki </h4>
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                        </html>
<p>On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world! </p>
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<p> <a href="https://2015.igem.org/wiki/index.php?title=Team:Washington&action=edit"> Click here to edit this page! </a></p>
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<p>See tips on how to edit your wiki on the <a href="https://2015.igem.org/TemplatesforTeams_Code_Documentation">Template Documentation</a> page.</p>
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<h4>Templates </h4>
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<p> This year we have created templates for teams to use freely. More information on how to use and edit the templates can be found on the
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<a href="https://2015.igem.org/TemplatesforTeams_Code_Documentation">Template Documentation </a> page.</p>
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<h4>Tips</h4>
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<p>This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: </p>
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<ul>
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<li>State your accomplishments! Tell people what you have achieved from the start. </li>
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<li>Be clear about what you are doing and how you plan to do this.</li>
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<li>You have a global audience! Consider the different backgrounds that your users come from.</li>
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<li>Make sure information is easy to find; nothing should be more than 3 clicks away.  </li>
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<li>Avoid using very small fonts and low contrast colors; information should be easy to read.  </li>
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<li>Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="https://2015.igem.org/Calendar_of_Events">iGEM 2015 calendar</a> </li>
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<li>Have lots of fun! </li>
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</ul>
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<h4>Inspiration</h4>
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<p> You can also view other team wikis for inspiration! Here are some examples:</p>
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<ul>
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<li> <a href="https://2014.igem.org/Team:SDU-Denmark/"> 2014 SDU Denmark </a> </li>
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<li> <a href="https://2014.igem.org/Team:Aalto-Helsinki">2014 Aalto-Helsinki</a> </li>
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<li> <a href="https://2014.igem.org/Team:LMU-Munich">2014 LMU-Munich</a> </li>
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<li> <a href="https://2014.igem.org/Team:Michigan"> 2014 Michigan</a></li>
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<li> <a href="https://2014.igem.org/Team:ITESM-Guadalajara">2014 ITESM-Guadalajara </a></li>
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<li> <a href="https://2014.igem.org/Team:SCU-China"> 2014 SCU-China </a></li>
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</ul>
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<h4> Uploading pictures and files </h4>
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<p> You can upload your pictures and files to the iGEM 2015 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name. <br />
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When you upload, set the "Destination Filename" to <code>Team:YourOfficialTeamName/NameOfFile.jpg</code>. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)</p>
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<a href="https://2015.igem.org/Special:Upload">CLICK HERE TO UPLOAD FILES</a>
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Latest revision as of 23:56, 18 September 2015





Lab on a Strip: Developing a Novel Platform for Yeast Biosensors

Project Overview

The Pacific Ocean is home to a wide range of marine life, including the food source of many filter-feeders, toxin-producing algae. When algal blooms are ingested by shellfish, the toxins produced by the algae are caught within shellfish tissue. Although these toxins are harmful to us, they aren’t to the shellfish, giving collectors no immediate sign of danger. Biotoxins are also just generally difficult to detect; contrary to popular belief, algal blooms are not always the striking crimson of “red tides.” Thus, blooms may not be discovered until after a poisoned shellfish is found. The Washington State Department of Health and commercial shellfish farmers conduct periodic surveys of local beaches to catch contaminations early, but these methods are costly, time-consuming, and not always effective. This can especially pose a dilemma for individual shellfish hunters, who do not have the resources to screen their shellfish for toxins. With current detection methods, the crowds swarming to Seattle’s famous Pike Place Market and popular raw oyster bars are constantly at risk.



We have developed a much cheaper diagnostic tool in which genetically-modified baker’s yeast is grown on a paper device and is able to produce an easy-to-read color output in the presence of a target molecule. Imagine if you could simply dip a sheet of paper into your bucket of shellfish, wait only (insert amount of time) and tell if your products are safe to consume. The proof-of-concept systems we’ve engineered detect the plant hormone auxin and the molecule theophylline. However, we’ve implemented a number of techniques to ensure the versatility of our systems thus, they can be easily modified and further developed to test for a wide variety of other molecules.

In the Auxin detection pathway, a DNA binding domain, a degron domain and a repressor domain are fused to suppress the expression of a reporter gene, LacZ. In the presence of Auxin, a plant hormone, along with a corresponding F-Box protein will lead to the fusion protein suppressing the reporter will be ubiquitinated allowing the reporter to be expressed.
In this design RNA aptamers are used to sense our target molecule, theophylline. Aptamers, unlike antibodies, can actually bind to virtually any molecule, allowing for a more versatile system. We’ve implemented a ribozyme switch which, when active, cleaves the mRNA code of our target sequence, hindering the production of GFP by default. However, in the presence of theophylline our switch becomes inactive, allowing for the expression of our target gene. This system is useful because it is faster-acting than more traditional expression pathways, and can be generalized to many other small molecules by changing the aptamer sequence.