Team:Pasteur Paris/Demonstrate
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