Difference between revisions of "Team:SHSBNU China/Improve"

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    <title>Improvements</title>
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    <script src="https://2018.igem.org/Template:SHSBNU_China/Sample_js?action=raw&ctype=text/javascript" type="text/javascript"></script>
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<h3>★  ALERT! </h3>
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<div style="height:5vw;" class="first_classfication" onmouseover="fcova()" onmouseout="fcoua()" id="fc_Project"><!--Project-->
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<p>This page is used by the judges to evaluate your team for the <a href="https://2018.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2018.igem.org/Judging/Awards"> award listed below</a>. </p>
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2018.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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<a class="fc_link" style="height:100%; width:100%;" href="https://2018.igem.org/Team:SHSBNU_China/Project">
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<h4 class="fc_content">Project</h4>
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<a class="fc_link" style="height:100%; width:100%;" href="https://2018.igem.org/Team:SHSBNU_China/Experiments">
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<h4 class="fc_content">Experiment</h4>
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<div style="height:5vw; padding: 0vw; align-self: center; vertical-align: middle; display: table; line-height: 3vw;" class="first_classfication" onmouseover="fcovh()" onmouseout="fcouh()" id="fc_Team"><!--Team-->
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<a class="fc_link" style="height:100%; width:100%;" href="https://2018.igem.org/Team:SHSBNU_China/Team">
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<h4 style="padding-right: 0.30vw">Team</h4>
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<a href="https://2018.igem.org/Team:SHSBNU_China">
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<p style="font-family: Serif;font-size: 3.5vw; font-weight: bold;text-align: center;margin: 0.5vw 0vw 0.5vw 0vw">Improve</p>
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<a class="snd_class" href="https://2018.igem.org/Team:SHSBNU_China/Improve#I">Overview</a>
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<a class="snd_class" href="https://2018.igem.org/Team:SHSBNU_China/Improve#ID">Improvement Details</a>
  
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<h1>Improve</h1>
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<a class="snd_class" href="https://2018.igem.org/Team:SHSBNU_China/Improve#R">References</a>
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<p>For teams seeking to improve upon a previous part or project, you should document all of your work on this page. Please remember to include all part measurement and characterization data on the part page on the Registry. Please include a link to your improved part on this page.</p>
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<h3>Gold Medal Criterion #2</h3>
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<p><b>Standard Tracks:</b> Create a new part that has a functional improvement upon an existing BioBrick part. The sequences of the new and existing parts must be different. You must perform experiments with both parts to demonstrate this improvement.  Document the experimental characterization on the Part's Main Page on the Registry for both the existing and new parts. Both the new and existing Main Page of each Part’s Registry entry must reference each other. Submit a sample of the new part to the Registry.
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The existing part must NOT be from your 2018 part number range and must be different from the part documented in bronze #4.
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<div id="articles">
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<h1>Improve</h1>
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<b>Special Tracks:</b> Improve the function of an existing iGEM project (that your current team did not originally create) and display your achievement on your wiki.</p>
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<h2 id="I">Improvements</h2>
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<div class="content">
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<p><img src="https://static.igem.org/mediawiki/2018/b/b9/T--SHSBNU_China--21000.png" style="width:75%"/></image></p>
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    Previously, there was only CsgA part uploaded in iGEM parts website. Our Team improved part <a href="http://parts.igem.org/Part:BBa_K1583000">Part BBa_K1583000</a> by adding a SpyTag sequence. It fused to gene <em>csgA</em>, enabling CotA laccase to be fixed onto the biofilm to form a covalent bond--SpyTag-SpyCatcher.
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<h2 id="ID">Improvement Details</h2>
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By using sfGFP-SpyCatcher protein, we tested the binding efficiency of the covalence SpyTag-SpyCatcher since sfGFP is a non-toxic and common used flurorescent protein. Gene <em>csgA</em> on the plasmid of pET28a was transferred into ΔMG1655 as control group. Gene <em>csgA – spytag</em> on the plasmid of pET28a was transferred into ΔMG1655 as experiment. The experiment was divided into 4 groups. The bateria in the Reaction Stock had neither <em>csgA</em> nor <em>SpyTag</em> in neither genome nor plasmid; the ones in Group 1 had <em>csgA</em> in the genome; the ones in Group 2 had <em>sfGFP</em> in the plasmid; the ones in Group 3 had <em>csgA-SpyTag</em> in the genome and the plasmid. The treatment of the experiment is followed by the procedure below:
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<p><img src="https://static.igem.org/mediawiki/2018/9/92/T--SHSBNU_China--improve1.jpg" style="width:75%"></p>
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<div style="width:20vw" class="content_pic_right">
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<img class="pictures" id = "21002" src="https://static.igem.org/mediawiki/2018/5/57/T--SHSBNU_China--21002.jpg"/>
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<p class="pic_text">The liquid after centrifugation was extracted and measured fluorescence. </p>
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As can be seen from the result, the experiment group showed the most decrease of sfGFP-SpyCatcher protein. The difference between the control group and experiment group was significant. Thus we could confirm our CsgA-SpyTag system is functional.</p>
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<p style="font-size:1vw;font-family:'Times New Roman';font-style:italic;color:#666666">SHSBNU_China 2018, Created by Azir</p>
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Revision as of 19:12, 17 October 2018

Improvements