Team:INSA-Lyon/Results

From 2014.igem.org

(Difference between revisions)
(Undo revision 364238 by Alex273 (talk))
Line 152: Line 152:
<div align="justify"><p>To adress biosafety issues linked with GMOs, we worked on destroying our bacteria after letting them grow in a biofilm. As the captured metal is extracellular and Curli proteins are very resistant to environmental changes, live bacteria are not needed for our biofilter. Our goal was to obtain a biomaterial made out of modified Curli able to chelate nickel. </p> <br/>
<div align="justify"><p>To adress biosafety issues linked with GMOs, we worked on destroying our bacteria after letting them grow in a biofilm. As the captured metal is extracellular and Curli proteins are very resistant to environmental changes, live bacteria are not needed for our biofilter. Our goal was to obtain a biomaterial made out of modified Curli able to chelate nickel. </p> <br/>
-
<p>To find the best way to degrade bacteria and DNA, the following protocol was used to test the influence of UV light and temperature separately : <br/>
+
<p>To find the best way to degrade bacteria and DNA, the following protocol was used to test the influence of UV light and temperature separately : <br/>
<ul>
<ul>
-
<li> Wells containing M63 cultures of strain 227 were put under UV light / at 60 or 70°C for different lengths of time. Well contents were then gradually transferred into Eppendorf and diluted (100, 300, 900 and 2700 fold).<br/>  
+
<li> Wells containing M63 cultures of strain 227 were put under UV light / at 60 or 70°C for different lengths of time. Well contents were then gradually transferred into Eppendorf and diluted (100, 300, 900 and 2700 fold).<br/>  
<li> LB plates (without antibiotic) corresponding to UV/temperature exposure times (+ one plate for control) were then spotted with s227 different concentrations in order to be able to count survival bacteria after incubation at 37°C.<br/>
<li> LB plates (without antibiotic) corresponding to UV/temperature exposure times (+ one plate for control) were then spotted with s227 different concentrations in order to be able to count survival bacteria after incubation at 37°C.<br/>
<li> Genomic DNA was extracted from s227 concentrated culture. From the solution obtained, Curli promoter(750 bp) was amplified by PCR with Q5 polymerase and designed primers. <br/>
<li> Genomic DNA was extracted from s227 concentrated culture. From the solution obtained, Curli promoter(750 bp) was amplified by PCR with Q5 polymerase and designed primers. <br/>
Line 160: Line 160:
</ul>
</ul>
-
<p><h6><i> UV light influence </i></h6></p><br/>
+
<p><h6> UV light influence </h6></p><br/>
 +
 
 +
<p><i>LB plates</i></p></br>
<p><div align="center"><table>
<p><div align="center"><table>
<tr>
<tr>
Line 186: Line 188:
No bacteria grew on LB plate after 15 minutes UV light exposure.<br/><b>Bacterian growth can be stopped this way. </b></p><br/>
No bacteria grew on LB plate after 15 minutes UV light exposure.<br/><b>Bacterian growth can be stopped this way. </b></p><br/>
 +
<p><i> DNA extraction</i></p>
<p><table><br/>
<p><table><br/>
<tr>
<tr>
Line 192: Line 195:
</table> <br/> Bacterian DNA seemed to be degraded after 10 min UV light exposure.<br/>&rArr; <b>In consequence, UV light can be used to destroy DNA.</b> </p><br/>
</table> <br/> Bacterian DNA seemed to be degraded after 10 min UV light exposure.<br/>&rArr; <b>In consequence, UV light can be used to destroy DNA.</b> </p><br/>
-
<p><table>
+
<p><div align="center"><table>
<tr>
<tr>
-
   <td><img src="https://static.igem.org/mediawiki/2014/e/ee/BacklightControl_UV.png" alt="Control plate" width="200 px"/></td>
+
   <td><img src="https://static.igem.org/mediawiki/2014/e/ee/BacklightControl_UV.png" alt="Control plate" width="300 px"/></td>
-
   <td><img src="https://static.igem.org/mediawiki/2014/b/bc/BacklightUV15.png" alt="15 min" width="200 px"/></td>
+
   <td><img src="https://static.igem.org/mediawiki/2014/b/bc/BacklightUV15.png" alt="15 min" width="300 px"/></td>
-
   <td><img src="https://static.igem.org/mediawiki/2014/4/4b/BacklightUV20.png" alt="20min" width="200 px"/></td>
+
   <td><img src="https://static.igem.org/mediawiki/2014/4/4b/BacklightUV20.png" alt="20min" width="300 px"/></td>
</tr>
</tr>
<tr>
<tr>
-
   <td><div align="center"><figcaption>Control plate</figcaption></td></div>
+
   <td><div align="center"><figcaption>Control plate</figcaption></div></td>
-
   <td><div align="center"><figcaption>15 min UV</figcaption></td></div>
+
   <td><div align="center"><figcaption>15 min UV</figcaption></div></td>
-
   <td><div align="center"><figcaption>20 min UV</figcaption></td></div>
+
   <td><div align="center"><figcaption>20 min UV</figcaption></div></td>
-
</tr>
+
</tr></table></div>
<br/> Still some green-colored bacteria could be seen after 20 min UV exposure. <br/>  
<br/> Still some green-colored bacteria could be seen after 20 min UV exposure. <br/>  
&rArr;<b>UV light isn’t enough to kill bacteria.</b></p><br/><br/>
&rArr;<b>UV light isn’t enough to kill bacteria.</b></p><br/><br/>

Revision as of 23:05, 17 October 2014

Curly'on - IGEM 2014 INSA-LYON

  • Curli characterization


  • Nickel chelation


  • Survival after UV and high temperature exposure


  • Promoter optimization and characterization