Team:EPF Lausanne/Project

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Contents

Overview

Our projects aims to help stopping the propagation of malaria. To do so, we chose to act on the mosquito. Indeed, if we prevent the mosquito from being infected by malaria, it won't be able to transmit the parasite to healthy humans. Our idea is to engineer asaia, a bacteria that is naturally present in the mosquito's intestinal tract. This bacteria would express an immunotoxin to kill the plasmodium and thus prevent the mosquito infection.

Movie

Coming soon!

Plan

After choosing our bacteria of interest, here Asaia, we decided on two main set of experiments.

Cloning

The first, of course, was the cloning part. We performed all the cloning in E.coli, and did experiments on Asaia transformation, growth and resistance to antibiotics in parallel.

Here are the parts we successfully cloned.

As E.Coli origin does not work in Asaia, we added an origin that was compatible with this bacteria. Gwen: where did we get this origin from? On the opposite, Asaia's origin does work in E.Coli, which is very useful for our sets of experiments.

dessin des "parts"

Asaia Characterization

We measured the doubling time of Asaia.

Graph growth curve2.jpg

We tested our bacteria with 9 differents antibiotics and measured the resistance with a paper filter test and a liquid culture test.


Graph Resistance2.jpg


Drosophila

In parallel, we did experiments to assess if Asaia was persistant in Drosophila melanogaster . In fact, the fly is a much easier organism to grow and work on, in opposition to mosquitoes which need to feed on animals to reproduce. Our experiment was set in 3 steps:

  • infection of the flies with GFP-Asaia*, and 3 other bacteria as controls
  • retrieval of the flies after 3h (control of infection), 24h, 48h
  • counting of the number of colonies for all time points and the 4 differents strains

The goal was to observe the persistance of Asaia in the Drosophila gut to see if it can be used as a model organism to study Asaia.

The results were negative for a the experience and we repeteated it 4 times.

graphe des drosophiles




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