Difference between revisions of "Team:Pasteur Paris/Demonstrate"
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<p><a href="#Nerves" class="link">Nerve Growth</a></p> | <p><a href="#Nerves" class="link">Nerve Growth</a></p> | ||
<p><a href="#Kill" class="link">Kill Switch</a></p> | <p><a href="#Kill" class="link">Kill Switch</a></p> | ||
| − | <p><a href="#Membrane" class="link">Membrane | + | <p><a href="#Membrane" class="link">Membrane</a></p> |
<p><a href="#Design" class="link">Design</a></p> | <p><a href="#Design" class="link">Design</a></p> | ||
</div> | </div> | ||
Revision as of 18:59, 16 October 2018
NERVE GROWTH FACTOR AND NEURON CULTURE
Achievements:
- Successfully cloned a part coding for secretion of NGF in pET43.1a and iGEM plasmid backbone pSB1C3, creating a new basic part Bba_K2616000
- Successfully sequenced Bba_K2616000 in pSB1C3 and sent to iGEM registry
- Successfully co-transform E. coli with plasmid secreting NGF and plasmid expressing the secretion system, creating bacteria capable of secreting NGF in the medium
- Successfully characterized production of NGF thanks to mass spectrometry
- Successfully observe axon growth in microfluidic chip in presence of commercial NGF
Next steps:
- Purify secreted NGF, and characterize its effects on neuron growth thanks to our microfluidic device
- Global proof of concept in a microfluidic device containing neurons in one of the chamber, and our engineered bacteria in the other
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
The sequence designed codes for two different proteins: CcdB toxin and CcdA antitoxin. The antitoxin production is under an constitutive promoter (PLac) and the toxin production under a thermosensitive one (PcspA).(Figure 19)
Figure 19: Cryo-death Kill-switch construction