Difference between revisions of "Team:Pasteur Paris/Demonstrate"

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             <div class="block title" id="Nerves">
 
             <div class="block title" id="Nerves">
 
                 <h1>NERVE GROWTH FACTOR AND NEURON CULTURE</h1>
 
                 <h1>NERVE GROWTH FACTOR AND NEURON CULTURE</h1>
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                 <i style="text-align: left;"><p>Achievements:<br>
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                 <p> We successfully cloned a part coding for secretion of NGF in pET43.1a and iGEM plasmid backbone pSB1C3, creating a new part <a href="http://parts.igem.org/Part:BBa_K2616000"style="font-weight: bold ; color:#85196a;"target="_blank"> Bba_K2616000 </a> and confirmed the production of proNGF by Western Blot and mass spectrometry. </p>
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                    <ul>
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                <p> We grew neurons on our self-made microfluidic chips ans successfully observe axon growth in the presence of commercial NGF</p>
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                        <li>Successfully cloned a part coding for secretion of NGF in pET43.1a and iGEM plasmid backbone pSB1C3, creating a new basic part <a href="http://parts.igem.org/Part:BBa_K2616000"style="font-weight: bold ; color:#85196a;"target="_blank"> Bba_K2616000 </a> </li>
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            </div>
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                        <li>Successfully sequenced <a href="http://parts.igem.org/Part:BBa_K2616000"style="font-weight: bold ; color:#85196a;"target="_blank"> Bba_K2616000 </a> in pSB1C3 and sent to iGEM registry </li>
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                        <li>Successfully co-transform E. coli with plasmid secreting NGF and plasmid expressing the secretion system, creating bacteria <b>capable of secreting NGF</b> in the medium</li>
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                        <li>Successfully characterized production of NGF thanks to mass spectrometry</li>
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                        <li>Successfully <b>observe axon growth</b> in microfluidic chip in presence of commercial NGF</li>
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                    </ul><br></p>
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                    <p>Next steps:<br>
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                    <ul>
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                        <li><b>Purify</b> secreted NGF, and characterize its effects on neuron growth thanks to our microfluidic device </li>
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                        <li><b>Global proof of concept</b> in a microfluidic device containing neurons in one of the chamber, and our engineered bacteria in the other</li>
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                    </ul>
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                </p></i>
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             </div>
 
             </div>
  

Revision as of 19:04, 16 October 2018

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NERVE GROWTH FACTOR AND NEURON CULTURE

We successfully cloned a part coding for secretion of NGF in pET43.1a and iGEM plasmid backbone pSB1C3, creating a new part Bba_K2616000 and confirmed the production of proNGF by Western Blot and mass spectrometry.

We grew neurons on our self-made microfluidic chips ans successfully observe axon growth in the presence of commercial NGF

KILL SWITCH

Achievements:

  • Successfully cloned a part coding for toxin/antitoxin (CcdB/CcdA) system in iGEM plasmid backbone, creating a new basic part Bba_K2616002
  • Successfully sequenced BBa_K2616002 in pSB1C3 and sent to iGEM registry
  • Successfully observe survival of our engineered bacteria at 25°C and 37°C and absence of growth at 18°C and 20°C, showing the efficiency of the kill switch

Next steps:

  • Find a system that kills bacteria when released in the environment rather than just stopping their growth

MEMBRANE BIOCOMPATIBILITY AND CONDUCTIVITY

DESIGN