Experiments Overview

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:

1 - Mimicking Vibrio sp. presence with an engineered E. coli

We engineered here E. coli to mimic for safety reason the presence of Vibrio species.

• Cloning:
• Integration in chassis:
• Validation:

2 - E. coli producing C8-CAI-1 molecules can be sensed by V. harveyi

In this module, we created synthetic communication between our engineered E. coli and V. harveyi.

• Cloning:
• Integration in chassis:
• Validation:

3 - Modification of V. harveyi to detect both C8-CAI-1 and CAI-1

Here, we present our first effective modification of V. harveyi and its perspectives for the project.

• Cloning:
• Integration in chassis:
• Validation:

4 - Establishing production of diacetyl to establish communication between prokaryotic and eukaryotic cells

This module is dedicated to the production of diacetyl by bacteria, as a signal relay toward yeast.

• Cloning:
• Integration in chassis:
• Validation:

5 - Diacetyl detection by Pichia pastoris

This presents are modification and first assays to engineered P. pastoris for diacetyl detection.

• Cloning:
• Integration in chassis:
• Validation:

6 - P. pastoris is able to produce functional antimicrobial peptides

Finally, this module described the efficiency of a yeast-produced crocodile AMP on V. harveyi.

• Cloning:
• Integration in chassis:
• Validation:

7 - Co-cultivate P. pastoris and V. harveyi is possible

Toward a realistic solution, we tested here how to co-culture how consortium microoganisms.

• Validation:

8 - Membrane permeability assay

We assessed the capacity of our microorganism-impermeable materials to let AMP go through.

• Validation: