Team:INSA-UPS France/Experiments
iGEM UPS-INSA Toulouse 2017
This Realisations section describes all our wet lab work, based on our strategy. We divided this section into the Clonings page and the Results page: the first shows how we integrated the parts of our Design into our chassis, and the second shows the experiments we did to further validate their functionnality. Both are presented for each of our eight modules:
We engineered here E. coli to mimic for safety reason the presence of Vibrio species. In this module, we created synthetic communication between our engineered E. coli and V. harveyi. Here, we present our first effective modification of V. harveyi and its perspectives for the project. This module is dedicated to the production of diacetyl by bacteria, as a signal relay toward yeast. This presents are modification and first assays to engineered P. pastoris for diacetyl detection. Finally, this module described the efficiency of a yeast-produced crocodile AMP on V. harveyi. Toward a realistic solution, we tested here how to co-culture our microbial consortium. We assessed the capacity of our microorganism-impermeable materials to let AMP go through. Experiments Overview
1 - Mimicking Vibrio sp. presence with an engineered E. coli
2 - E. coli producing C8-CAI-1 molecules can be sensed by V. harveyi
3 - Modification of V. harveyi to detect both C8-CAI-1 and CAI-1
4 - Establishing production of diacetyl to establish communication between prokaryotic and eukaryotic cells
5 - Diacetyl detection by Pichia pastoris
6 - P. pastoris is able to produce functional antimicrobial peptides
7 - Co-cultivate P. pastoris and V. harveyi is possible
8 - Membrane permeability assay
Realisations pages
Overview
Notebook
Clonings
Results
Contribution
Protocols
Safety
