Difference between revisions of "Team:Paris Bettencourt/Proof"

 
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<h2 class="red">Proof-of-concept - developing fabric-specific bindig domains (Fabric Binding Domains)</h2>
  
 
<p>
 
<p>
−
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.  
+
As our proof-of-concept we show that the Fabric Binding Domains (FBD) that we developed in this project work, by having affinity and specificity for a given type of fabric. <br>
 +
The complete list of FBDs follows. A more broader description of each FBD can be found in their Biobricks' page.
 
</p>
 
</p>
  
 +
<table id=table_style>
 +
  <tr style="background-color:#438CEE">
 +
    <th Colspan="4" style="background-color:#8B1813; color:#FFFFFF; font-size:20px;">Enzymes for Stain Digestion</th>
 +
  </tr>
 +
  <tr style="background-color:#138B3F">
 +
    <th style="background-color:#DDDDDD">BioBrick ID</th>
 +
    <th style="background-color:#DDDDDD">Enzyme name</th>
 +
    <th style="background-color:#DDDDDD">Description</th>
 +
    <th style="background-color:#DDDDDD">Reference</th>
 +
  </tr>
  
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<h4> What should we do for our proof of concept? </h4>
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<p>  
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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!
+
    <tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K2043011">BBa_K2043011</a></td>
 +
    <td>GFP-FBD2</td>
 +
    <td style="text-align: justify">GFP gene, codon optimised for <i>E. coli</i> and fused in the N-terminal with FBD2 </td>
 +
    <td style="text-align: justify"</td>
 +
  </tr>
 +
 
 +
    <tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K2043012">BBa_K2043012</a></td>
 +
    <td>GFP-FBD5</td>
 +
    <td style="text-align: justify">GFP gene, codon optimised for <i>E. coli</i> and fused in the N-terminal with FBD5 </td>
 +
    <td style="text-align: justify"></td>
 +
  </tr>
 +
 
 +
    <tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K2043013">BBa_K2043013</a></td>
 +
    <td>GFP-FBD6</td>
 +
    <td style="text-align: justify">GFP gene, codon optimised for <i>E. coli</i> and fused in the N-terminal with FBD6</td>
 +
    <td style="text-align: justify"></td>
 +
  </tr>
 +
 
 +
    <tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K2043014">BBa_K2043014</a></td>
 +
    <td>GFP-FBD7</td>
 +
    <td style="text-align: justify">GFP gene, codon optimised for <i>E. coli</i> and fused in the N-terminal with FBD7 </td>
 +
    <td style="text-align: justify"></td>
 +
  </tr>
 +
 
 +
 
 +
    <tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K2043016">BBa_K2043016</a></td>
 +
    <td>GFP-FBD9</td>
 +
    <td style="text-align: justify">GFP gene, codon optimised for <i>E. coli</i> and fused in the N-terminal with FBD9 </td>
 +
    <td style="text-align: justify"></td>
 +
  </tr>
 +
 
 +
    <tr>
 +
    <td><a href="http://parts.igem.org/Part:BBa_K2043017">BBa_K2043017</a></td>
 +
    <td>GFP-FBD10</td>
 +
    <td style="text-align: justify">GFP gene, codon optimised for <i>E. coli</i> and fused in the N-terminal with FBD10 </td>
 +
    <td style="text-align: justify"></td>
 +
  </tr>
 +
</table>
 +
 
 +
<p>
 +
The following figure is proof that the FBDs work. Our results show that each FBD has an individual relationship with each of the fabrics, some having affinity to many, other showing specificity to a given fabric.
 
</p>
 
</p>
  
 +
<div id="figurebox">
 +
<img src="https://static.igem.org/mediawiki/2016/f/fe/Paris_Bettencourt-enzyme_heatmaps.jpg" alt="Quercetin strains degradation" style="width:800px;">
 +
<p>
 +
<b>Figure 5: Analysis of FBD-GFP fusion proteins. </b>With this image we show that the GFP activity was not affected by the fusion between the FBDs and the GFP. CBD stands for the +control, cellulose binding domain from the Biobrick registry BBa_K1321357. <br>
 +
Cell extract from strains expressing our fusion proteins was incubated overnight with the fabrics. Afterwards two washes of the fabrics were performed. Water, PBS, BSA 5%, Ethanol 70% and Catechol correspond to the washing solutions we used to remove the non-bound GFP, as well as to test the binding strength of the different peptides. The data displayed corresponds to the values after the final wash, normalised using the values from the first wash. <br>
 +
The intensity of the colour corresponds to the GFP signal measured at that point. <br
 +
Each heatmap corresponds to the results obtained with different fabrics. Results were obtained using the final 96-wells-based assay designed by our Assay team, using untreated, unbleached cotton, linen, silk and wool.
 +
</p>
 
</div>
 
</div>
  
 +
<div style="margin-top:20px; margin-bottom:20px"></div>
  
 +
<div class="panel" >
 +
  <a href="https://2016.igem.org/Team:Paris_Bettencourt/Results" title="Results">
 +
    <div id="pspanel" class="subpanel2"  onmouseover="chgtrans(this)">
 +
      <img class="narrowimg" src="https://static.igem.org/mediawiki/2016/d/db/T--Paris_Bettencourt--results_icon.jpg" width="400px" height="300px"/>
 +
      <div class="titlebox">
 +
<center><img src="https://static.igem.org/mediawiki/2016/3/3e/T--Paris_Bettencourt--results_logo.png" style="height:90px;margin-top:-30px;"/></center>
 +
<div style="width:60%;margin-left:20%;margin-bottom:0px;"><hr></div>
 +
Results
 +
      </div>
 +
    </div>
 +
  </a>
  
 +
  <a href="https://2016.igem.org/Team:Paris_Bettencourt/Medal_Requirements" title="Medal Requirements">
 +
    <div id="pspanel" class="subpanel2"  onmouseover="chgtrans(this)">
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      <img class="narrowimg" src="https://static.igem.org/mediawiki/2016/b/bb/T--Paris_Bettencourt--medal_icon.jpg" width="400px" height="300px"/>
 +
      <div class="titlebox">
 +
<center><img src="https://static.igem.org/mediawiki/2016/2/2c/T--Paris_Bettencourt--medal_logo.png" style="height:90px;margin-top:-30px;"/></center>
 +
<div style="width:60%;margin-left:20%;margin-bottom:0px;"><hr></div>
 +
Medal Requirements
 +
      </div>
 +
    </div>
 +
  </a>
 +
 +
  <a href="https://2016.igem.org/Team:Paris_Bettencourt/Parts" title="Parts">
 +
    <div id="pspanel" class="subpanel2"  onmouseover="chgtrans(this)">
 +
      <img class="narrowimg" src="https://static.igem.org/mediawiki/2016/0/03/T--Paris_Bettencourt--parts_icon.jpg" width="400px" height="300px"/>
 +
      <div class="titlebox">
 +
<center><img src="https://static.igem.org/mediawiki/2016/3/3a/T--Paris_Bettencourt--biobrick_logo.png" style="height:90px;margin-top:-30px;"/></center>
 +
<div style="width:60%;margin-left:20%;margin-bottom:0px;"><hr></div>
 +
Parts
 +
      </div>
 +
    </div>
 +
  </a>
 +
 +
  <a href="https://2016.igem.org/Team:Paris_Bettencourt/Interlab_Study" title="Interlab Study">
 +
    <div id="pspanel" class="subpanel2"  onmouseover="chgtrans(this)">
 +
      <img class="narrowimg" src="https://static.igem.org/mediawiki/2016/2/2e/T--Paris_Bettencourt--interlab_icon.jpg" width="400px" height="300px"/>
 +
      <div class="titlebox">
 +
<center><img src="https://static.igem.org/mediawiki/2016/8/85/T--Paris_Bettencourt--interlab_logo.png" style="height:90px;margin-top:-30px;"/></center>
 +
<div style="width:60%;margin-left:20%;margin-bottom:0px;"><hr></div>
 +
Interlab Study
 +
      </div>
 +
    </div>
 +
  </a>
 
</div>
 
</div>
 +
</div>
 +
</div>
 +
 +
</body>
 +
  
 
</html>
 
</html>
 +
{{Paris_Bettencourt/Footer}}

Latest revision as of 00:37, 20 October 2016


Proof-of-concept - developing fabric-specific bindig domains (Fabric Binding Domains)

As our proof-of-concept we show that the Fabric Binding Domains (FBD) that we developed in this project work, by having affinity and specificity for a given type of fabric.
The complete list of FBDs follows. A more broader description of each FBD can be found in their Biobricks' page.

Enzymes for Stain Digestion
BioBrick ID Enzyme name Description Reference
BBa_K2043011 GFP-FBD2 GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD2
BBa_K2043012 GFP-FBD5 GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD5
BBa_K2043013 GFP-FBD6 GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD6
BBa_K2043014 GFP-FBD7 GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD7
BBa_K2043016 GFP-FBD9 GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD9
BBa_K2043017 GFP-FBD10 GFP gene, codon optimised for E. coli and fused in the N-terminal with FBD10

The following figure is proof that the FBDs work. Our results show that each FBD has an individual relationship with each of the fabrics, some having affinity to many, other showing specificity to a given fabric.

Quercetin strains degradation

Figure 5: Analysis of FBD-GFP fusion proteins. With this image we show that the GFP activity was not affected by the fusion between the FBDs and the GFP. CBD stands for the +control, cellulose binding domain from the Biobrick registry BBa_K1321357.
Cell extract from strains expressing our fusion proteins was incubated overnight with the fabrics. Afterwards two washes of the fabrics were performed. Water, PBS, BSA 5%, Ethanol 70% and Catechol correspond to the washing solutions we used to remove the non-bound GFP, as well as to test the binding strength of the different peptides. The data displayed corresponds to the values after the final wash, normalised using the values from the first wash.
The intensity of the colour corresponds to the GFP signal measured at that point.

Results
Medal Requirements
Parts
Interlab Study
Centre for Research and Interdisciplinarity (CRI)
Faculty of Medicine Cochin Port-Royal, South wing, 2nd floor
Paris Descartes University
24, rue du Faubourg Saint Jacques
75014 Paris, France
+33 1 44 41 25 22/25
igem2016parisbettencourt@gmail.com
2016.igem.org