Difference between revisions of "Team:Goettingen/Proof"

 
(29 intermediate revisions by 2 users not shown)
Line 1: Line 1:
 
{{Goettingen/CSS}}
 
{{Goettingen/CSS}}
 +
  
 
<html>
 
<html>
  
−
<!-- *************************** SIDEBAR **************************** -->
+
<head>
  
−
<div class="sidebar">
 
−
<br>
 
  
−
<div class="sidebar">
+
</head>
  
−
<!-- ****** CONTACT ***** -->
 
  
−
<h1 style="text-align: center;">Contact</h1>
 
−
<br>
 
−
<ul>
 
−
<li><a href="https://www.facebook.com/iGEM-Team-Goettingen-687557807970778"><img src="https://static.igem.org/mediawiki/2016/3/39/T--Goettingen--Facebook_icon.png" width="32px"/>&nbsp; Facebook</a></li>
 
−
<li><a href="mailto:igem2016@gwdg.de"><img src="https://static.igem.org/mediawiki/2016/4/42/T--Goettingen--Mail_icon.png" width="32px"/>&nbsp; E-Mail</a></li>
 
−
<li><a href="http://igem2016.uni-goettingen.de/"><img src="https://static.igem.org/mediawiki/2016/f/fb/T--Goettingen--Web_icon.png" width="32px"/>&nbsp; Homepage</a></li>
 
−
<br>
 
−
<li><a href="http://appmibio.uni-goettingen.de/"><img src="https://static.igem.org/mediawiki/2016/d/d0/T--Goettingen--Post.png" width="32px"/>&nbsp; Address</a></li>
 
−
</ul>
 
−
</div>
 
  
−
<a href="http://appmibio.uni-goettingen.de/">
 
−
<div class="sidebaraddress">
 
−
<font size="-1">
 
−
iGEM Team Göttingen 2016<br>
 
−
Institut für Mikrobiologie und Genetik<br>
 
−
AG Prof. Dr. Rolf Daniel<br>
 
−
Grisebachstr. 8<br>
 
−
37077 Göttingen<br>
 
−
</font>
 
−
<centering>
 
−
<br>
 
−
</div>
 
−
</a>
 
  
 +
<body>
  
−
<div class="sidebar">
 
  
−
<!-- *** NAVIGATION *** -->
 
  
−
<h1 style="text-align: center;">Navigation</h1>
 
  
−
<br>
 
  
−
<div class="menu_wrapper" >
+
<div id="pageContent">
  
−
<ul id="accordion" class="accordion">
 
  
−
<li class="menu_item"> <a href="https://2016.igem.org/Team:Goettingen">Home </a> </li>
 
  
  
−
<li class="menu_item"> <div class="icon plus"></div> Team
 
−
<ul class="submenu">
 
−
<li> <a href=" https://2016.igem.org/Team:Goettingen/Team"> Team  </a> </li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Collaborations">Collaborations</a> </li>
 
−
                    </ul>
 
−
                </li>
 
  
−
<li class="menu_item" style="background-color: #e7e7e7;"> <div class="icon plus"></div> <b>Project</b>
 
−
<ul class="submenu">
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Description">Description</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Design">Design</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Experiments"> Experiments </a></li>
 
−
<li> <a style="background-color: #d8d8d8;" href="https://2016.igem.org/Team:Goettingen/Proof"><b>Proof of Concept</b></a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Demonstrate">Demonstrate</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Results"> Results </a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Notebook"> Notebook </a></li>
 
−
                    </ul>
 
−
                </li>
 
  
−
<li class="menu_item"> <div class="icon plus"></div> Parts
+
<!-- ***************** MAIN PAGE ************************ -->
−
<ul class="submenu">
+
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Parts">Parts</a></li>
+
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Basic_Part">Basic Parts </a></li>
+
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Composite_Part">Composite Parts</a></li>
+
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Part_Collection">Part Collection</a></li>
+
−
                    </ul>
+
−
                </li>
+
  
 +
<!-- page content plus sidebar -->
  
−
<li class="menu_item"> <a href="https://2016.igem.org/Team:Goettingen/Safety">Safety</a> </li>
+
  <div class="row">
 +
  <div class="col-md-12">  
 +
 
  
  
−
<li class="menu_item"> <a href="https://2016.igem.org/Team:Goettingen/Attributions">Attributions</a></li>
 
  
  
−
<li class="menu_item"> <div class="icon plus"></div> Human Practices
 
−
<ul class="submenu">
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Human_Practices">Human Practices</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/HP/Silver">Silver</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/HP/Gold">Gold</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Integrated_Practices">Integrated Practices</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Engagement">Engagement</a></li>
 
−
                    </ul>
 
−
                </li>
 
  
  
−
<li class="menu_item"> <div class="icon plus"></div> Awards
 
−
<ul class="submenu">
 
−
<li><a href="https://2016.igem.org/Team:Goettingen/Entrepreneurship">Entrepreneurship</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Hardware">Hardware</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Software">Software</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Measurement">Measurement</a></li>
 
−
<li> <a href="https://2016.igem.org/Team:Goettingen/Model">Model</a></li>
 
  
−
                    </ul>
+
<!-- ***************** PAGE CONTENT ************************ -->
−
                </li>
+
−
</ul>
+
  
−
</div>
 
−
</div>
 
  
 +
  <div class="row">
 +
  <div class="col-md-9"> <!-- page content -->
  
−
<div class="sidebar" >
 
  
 +
<!-- heading -->
  
−
<!-- *** SPONSORS *** -->
+
<div class="heading">
 +
 
 +
<h1> Proof of Concept </h1>
 +
 
 +
  </div> <!-- close text div -->
  
  
−
<br>  
+
<!-- actual content -->
  
−
<h1 style="text-align: center;">Sponsors</h1>
+
<div class="row">  
 +
<div class="col-md-12">
  
  
 +
<div class="text">
  
−
<div class="sidebarsponsors" >
 
  
−
<a href="https://www.bbraun.de/de.html"><img class="wp-image-346 aligncenter" style="width: 100%; height: 100%;" src="https://static.igem.org/mediawiki/2016/5/5c/T--Goettingen--bbraun_small.png" alt="bbraun" /></a>
+
<p>Our aim was to transport Vitamin B<sub>12</sub>, a substance which has no known cellular exporter in nature, through the inner membrane out of the cytoplasm. Our different B<sub>12</sub> Synporter proteins consist of a TorA signal peptide that mediates export through the Twin Arginine Translocation (Tat) system, linked to a B<sub>12</sub>-binding protein domain. This construct was produced, together with B<sub>12</sub>, by the B<sub>12</sub> prototrophic enterobacteria <em>R. planticola</em> and <em>S. blattae</em>.<p>
  
 +
<hr>
 +
<img class="photo" src="https://static.igem.org/mediawiki/2016/5/54/T--Goettingen--Synporter.png" style="width:80%;"/>
 +
<hr>
  
  
−
<a href="http://www.mn-net.com/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/1/1c/T--Goettingen--m-n_small.png" alt="Macherey-Nagel" /></a><a href="http://www.zymoresearch.com/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/c/ce/T--Goettingen--zymo_small.png" alt="zymo" /></a>
+
<p>We were able to detect the Synporter proteins in the periplasm of the two organisms (see <a href="https://2016.igem.org/Team:Goettingen/Results#western">expression test</a>, proving that the Tat mediated translocation was successful. Moreover, most important, we could also prove a significantly increased overall B<sub>12</sub> production in the cells expressing the Synporter protein, compared to the cells containing an empty plasmid (see <a href="https://2016.igem.org/Team:Goettingen/Proof#final">B<sub>12</sub> assay</a>). This increased production is mainly due to the fact that the cells have to compensate for the B<sub>12</sub> leaving the cytoplasm bound to the Synporter. Thus, it proves the functionality of our system.</p>
 +
<p>This is the first time reported that a compound of such a high-molecular weight as B<sub>12</sub> was translocated through a cellular membrane without a native exporter. Therefore, our Synporter is truly innovative!</p>
  
−
<a href="http://www.snapgene.com/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/1/17/T--Goettingen--snapgene_small.png" alt="snapgene" /></a><a href="http://www.geneious.com/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/1/14/T--Goettingen--genious_small.png" alt="genious_logo" />
 
  
−
<a href="http://www.gatc-biotech.com/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/7/79/T--Goettingen--gatc_small.png" alt="gatc" /></a><a href="http://www.carlroth.com/de/de"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/a/ae/T--Goettingen--roth_small.png" alt="roth" /></a>
+
<p>Vitamin B<sub>12</sub> can also be regarded as a prime example for how highly valuable compounds can be gained from production organisms. In principle, a Synporter should work with basically any desired chemical compound, if a protein counterpart exists which can be used as Synporter binding domain. This concept could be applied for the production of all natural chemicals which are synthesized by microorganisms, but not exported due to the lack of a native cellular exporter. Furthermore, this principle could also be applied for the production of artificial molecules which can be produced by synthetic metabolic engineering in a cell. Using our Synporter approach, no cell lysis would be required any more for the industrial production. This concept has the potential to make the production process significantly easier and cheaper.</p>
  
−
<a href="http://idtdna.com"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/7/78/T--Goettingen--idt_small.png" alt="idt" /></a><a href="http://www.uni-goettingen.de/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/a/a4/T--Goettingen--uni-goe_small.png" alt="uni-goe" /></a>
+
<hr>
−
 
+
−
<a href="http://www.uni-goettingen.de/en/115976.html"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/4/40/T--Goettingen--gzmb_small.png" alt="gzmb" /></a><a href="http://appmibio.uni-goettingen.de/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/b/bf/T--Goettingen--appmibio_small.png" alt="appmibio" /></a>
+
  
 +
<img id="final" src="https://static.igem.org/mediawiki/2016/4/45/T--Goettingen--Result_Graph.png" class="photo" style="width:80%;" />
 +
<div class="caption">
 +
<p><b>Fig. 5: Final results of the photometric assay.</b> The plot shows the quantity of B<sub>12</sub> per amount of protein, in the periplasmic fraction and the cytoplasmic fraction.</p>
 
</div>
 
</div>
 +
<hr>
 +
 +
  
−
</div>
 
  
 
</div>
 
</div>
  
−
<!-- ************************************************************** -->
 
  
  
 +
</div> <!-- close row div -->
 +
</div> <!-- close col-md-9 div -->
  
−
<div class="text">
 
−
<div class="column full_size judges-will-not-evaluate">
 
−
<h3>★  ALERT! </h3>
 
−
<p>This page is used by the judges to evaluate your team for the <a href="https://2016.igem.org/Judging/Medals">gold medal criterion for proof of concept</a>. </p>
 
  
  
−
<p> Delete this box in order to be evaluated for this medal. See more information at <a href="https://2016.igem.org/Judging/Pages_for_Awards/Instructions"> Instructions for Pages for awards</a>.</p>
+
<!-- ************************************************************************************************************************************************************ -->
−
</div>
+
  
  
Line 166: Line 104:
  
  
−
<div class="column full_size">
 
  
  
−
<p>
+
</div> <!-- close col-md-12 div-->
−
iGEM teams are great at making things work! We value teams not only doing an incredible job with theoretical models and experiments, but also in taking the first steps to make their project real.
+
−
</p>
+
  
  
−
<h4> What should we do for our proof of concept? </h4>
+
 
−
<p>  
+
 
−
You can assemble a device from BioBricks and show it works. You could build some equipment if you're competing for the hardware award. You can create a working model of your software for the software award. Please note that this not an exhaustive list of activities you can do to fulfill the gold medal criterion. As always, your aim is to impress the judges!
+
 
−
</p>
+
 
 +
  <div class="col-md-3"> <!-- sidebar -->
 +
 
 +
 
 +
 
 +
 
 +
 
 +
<!-- *************** SIDEBAR ******************* -->
 +
 
 +
 
 +
  <div class="sidebar">
 +
  <br>
 +
    <h3 style="text-align: center;">Contact</h3>
 +
  <div class="sidebaraddress">
 +
 
 +
  <!-- *** CONTACT *** -->
 +
 
 +
 
 +
  <br>
 +
  <ul>
 +
  <li><a href="https://www.facebook.com/iGEM-Team-Goettingen-687557807970778"><img src="https://static.igem.org/mediawiki/2016/3/39/T--Goettingen--Facebook_icon.png" width="32px"/>&nbsp; Facebook</a></li>
 +
<li><a href="https://twitter.com/iGEM_Goettingen"><img src="https://static.igem.org/mediawiki/2016/c/c8/T--Goettingen--Twitter_icon.png" width="32px"/>&nbsp; Twitter</a></li>
 +
  <li><a href="mailto:igem2016@gwdg.de"><img src="https://static.igem.org/mediawiki/2016/4/42/T--Goettingen--Mail_icon.png" width="32px"/>&nbsp; E-Mail</a></li>
 +
  <li><a href="http://igem2016.uni-goettingen.de/"><img src="https://static.igem.org/mediawiki/2016/f/fb/T--Goettingen--Web_icon.png" width="32px"/>&nbsp; Homepage</a></li>
 +
  <br>
 +
  <li><a href="http://appmibio.uni-goettingen.de/"><img src="https://static.igem.org/mediawiki/2016/d/d0/T--Goettingen--Post.png" width="32px"/>&nbsp; Address</a></li>
 +
  </ul>
 +
  </div>
 +
 
 +
  <a href="http://appmibio.uni-goettingen.de/">
 +
  <div class="sidebaraddress" style="padding-left:42px; margin-top:10px;">
 +
  <font size="-1">
 +
  iGEM Team Göttingen 2016<br>
 +
  Institut für Mikrobiologie <br>
 +
  und Genetik<br>
 +
  AG Prof. Dr. Rolf Daniel<br>
 +
  Grisebachstr. 8<br>
 +
  37077 Göttingen<br>
 +
  </font>
 +
  <br>
 +
  </div>
 +
  </a>
 +
 
 +
 
 +
  <div class="sidebar_navigation">
 +
 
 +
  <!-- *** NAVIGATION *** -->
 +
 
 +
  <h3 style="text-align: center;">Navigation</h3>
 +
 
 +
  <br>
 +
 
 +
  <div class="menu_wrapper">
 +
 
 +
  <ul id="accordion" class="accordion">
 +
 
 +
  <li class="menu_item"> <a href="https://2016.igem.org/Team:Goettingen">Home </a> </li>
 +
 
 +
 
 +
  <li class="menu_item"> <div class="icon plus"></div> Team
 +
  <ul class="submenu">
 +
  <li> <a href=" https://2016.igem.org/Team:Goettingen/Team"> Team  </a> </li>
 +
  <li> <a href="https://2016.igem.org/Team:Goettingen/Collaborations">Collaborations</a> </li>
 +
  </ul>
 +
  </li>
 +
 
 +
<li class="menu_item" style="background-color: #e7e7e7;" > <div class="icon plus"></div> <b>Project</b>
 +
<ul class="submenu">
 +
<li> <a  href="https://2016.igem.org/Team:Goettingen/Description">Description</a></li>
 +
<li> <a  href="https://2016.igem.org/Team:Goettingen/Experiments">Experiments</a></li>
 +
<li> <a style="background-color: #D8D8D8;" href="https://2016.igem.org/Team:Goettingen/Proof"><b>Proof of Concept</b></a></li>
 +
<li> <a href="https://2016.igem.org/Team:Goettingen/Results"> Results </a></li>
 +
<li> <a href="https://2016.igem.org/Team:Goettingen/Notebook"> Notebook </a></li>
 +
  </ul>
 +
  </li>
 +
 
 +
  <li class="menu_item"> <div class="icon plus"></div> Parts
 +
  <ul class="submenu">
 +
  <li> <a href="https://2016.igem.org/Team:Goettingen/Parts">Parts</a></li>
 +
  <li> <a href="https://2016.igem.org/Team:Goettingen/Basic_Part">Basic Parts </a></li>
 +
  </ul>
 +
  </li>
 +
 
 +
 
 +
  <li class="menu_item"> <a href="https://2016.igem.org/Team:Goettingen/Safety">Safety</a> </li>
 +
 
 +
 
 +
  <li class="menu_item"> <a href="https://2016.igem.org/Team:Goettingen/Attributions">Attributions</a></li>
 +
 
 +
 
 +
  <li class="menu_item"> <div class="icon plus"></div> Human Practices
 +
  <ul class="submenu">
 +
  <li> <a href="https://2016.igem.org/Team:Goettingen/Human_Practices">Human Practices</a></li>
 +
  <li> <a href="https://2016.igem.org/Team:Goettingen/HP/Silver">HP on the Road</a></li>
 +
  <li> <a href="https://2016.igem.org/Team:Goettingen/HP/Gold">HP on the Web</a></li>
 +
  <li> <a href="https://2016.igem.org/Team:Goettingen/Engagement">Engagement</a></li>
 +
  </ul>
 +
  </li>
 +
 
 +
 
 +
  </ul>
 +
 
 +
  </div>
 +
  </div>
 +
 
 +
 
 +
 
 +
 
 +
 
 +
  <!-- *** SPONSORS *** -->
 +
 
 +
  <div class="sidebar" >
 +
  <br>
 +
  <h3 style="text-align: center;">Sponsors</h3>
 +
 
 +
 
 +
  <div class="sidebarsponsors" >
 +
 
 +
  <a href="https://www.bbraun.de/de.html"><img style="width: 100%; height: 100%; margin-bottom:0;" src="https://static.igem.org/mediawiki/2016/6/6d/T--Goettingen--sartorius_small.png" alt="sartorius" /></a>
 +
 
 +
 
 +
  <a href="https://www.bbraun.de/de.html"><img style="width: 100%; height: 100%; margin-top:0;" src="https://static.igem.org/mediawiki/2016/5/5c/T--Goettingen--bbraun_small.png" alt="bbraun" /></a>
 +
 
 +
 
 +
  <a href="http://www.mn-net.com/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/1/1c/T--Goettingen--m-n_small.png" alt="Macherey-Nagel" /></a><a href="http://www.zymoresearch.com/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/c/ce/T--Goettingen--zymo_small.png" alt="zymo" /></a>
 +
 
 +
  <a href="http://www.snapgene.com/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/1/17/T--Goettingen--snapgene_small.png" alt="snapgene" /></a><a href="http://www.geneious.com/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/1/14/T--Goettingen--genious_small.png" alt="genious_logo" />
 +
 
 +
  <a href="http://www.gatc-biotech.com/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/7/79/T--Goettingen--gatc_small.png" alt="gatc" /></a><a href="http://www.carlroth.com/de/de"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/a/ae/T--Goettingen--roth_small.png" alt="roth" /></a>
 +
 
 +
  <a href="http://idtdna.com"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/7/78/T--Goettingen--idt_small.png" alt="idt" /></a><a href="http://www.uni-goettingen.de/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/a/a4/T--Goettingen--uni-goe_small.png" alt="uni-goe" /></a>
 +
 
 +
  <a href="http://www.uni-goettingen.de/en/115976.html"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/4/40/T--Goettingen--gzmb_small.png" alt="gzmb" /></a><a href="http://appmibio.uni-goettingen.de/"><img style="width: 50%; height: 50%;" src="https://static.igem.org/mediawiki/2016/b/bf/T--Goettingen--appmibio_small.png" alt="appmibio" /></a>
 +
 
 +
  </div>
 +
 
 +
  </div>
 +
 
 +
  </div>
 +
 
 +
 
 +
  <!-- **************************** -->
  
 
</div>
 
</div>
  
 
</div>
 
</div>
 +
 +
  
 
</div>
 
</div>
  
−
</html>
+
 
 +
 
 +
  </body>
 +
  </html>

Latest revision as of 01:44, 20 October 2016


Proof of Concept

Our aim was to transport Vitamin B12, a substance which has no known cellular exporter in nature, through the inner membrane out of the cytoplasm. Our different B12 Synporter proteins consist of a TorA signal peptide that mediates export through the Twin Arginine Translocation (Tat) system, linked to a B12-binding protein domain. This construct was produced, together with B12, by the B12 prototrophic enterobacteria R. planticola and S. blattae.

We were able to detect the Synporter proteins in the periplasm of the two organisms (see expression test, proving that the Tat mediated translocation was successful. Moreover, most important, we could also prove a significantly increased overall B12 production in the cells expressing the Synporter protein, compared to the cells containing an empty plasmid (see B12 assay). This increased production is mainly due to the fact that the cells have to compensate for the B12 leaving the cytoplasm bound to the Synporter. Thus, it proves the functionality of our system.

This is the first time reported that a compound of such a high-molecular weight as B12 was translocated through a cellular membrane without a native exporter. Therefore, our Synporter is truly innovative!

Vitamin B12 can also be regarded as a prime example for how highly valuable compounds can be gained from production organisms. In principle, a Synporter should work with basically any desired chemical compound, if a protein counterpart exists which can be used as Synporter binding domain. This concept could be applied for the production of all natural chemicals which are synthesized by microorganisms, but not exported due to the lack of a native cellular exporter. Furthermore, this principle could also be applied for the production of artificial molecules which can be produced by synthetic metabolic engineering in a cell. Using our Synporter approach, no cell lysis would be required any more for the industrial production. This concept has the potential to make the production process significantly easier and cheaper.

Fig. 5: Final results of the photometric assay. The plot shows the quantity of B12 per amount of protein, in the periplasmic fraction and the cytoplasmic fraction.


Contact


iGEM Team Göttingen 2016
Institut für Mikrobiologie
und Genetik
AG Prof. Dr. Rolf Daniel
Grisebachstr. 8
37077 Göttingen

Navigation



Sponsors

sartorius bbraun Macherey-Nagelzymo snapgenegenious_logo gatcroth idtuni-goe gzmbappmibio