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Contents 1 Overall project 2 Project Details 2.1 Step 0 2.1.1 Step 0.1 2.1.2 Step 0.2 3 Results

Overall project

We started to work on one of the greatest discovery in cell biology: the existence of nanotubes in Bacillus subtilis!
We decided to take advantage of an article published by Dubey and Ben-Yehuda [http://bms.ucsf.edu/sites/ucsf-bms.ixm.ca/files/marjordan_06022011.pdf] where they show an extraordinary new form of communication between Bacillus subtilis cells and even exchanges with E. coli. In the first place, we want to prove de novo what the authors found. Although some microscopy images prove solidly the existence of these said nanotubes, we aim at obtaining our own results as well as gaining confidence in the manipulation of this bacteria. For this purpose, we started re-doing some experiences that were done in the articles notably the transient antibiotic resistance.
Our second aim is to caracterize the nanotubes: what passes through them and with which diffusion times. We will examine if RNA, proteins of different sizes and metabolites can pass through and with which ease and rate. For that purpose, we are going to engineer thanks to Synthetic Biology [http://en.wikipedia.org/wiki/BioBrick BioBricks] the following general design: an emitter cell would produce a messenger (RNA, protein etc.) that would then pass through the nanotubes and into the receiver cell. The latter, would then have specific promoters that would induce an amplification system that would in turn trigger a detection mechanism (fluroescence, others). This second aim will be implemented both at an intra-species level (subtilis to subtilis) and then to an inter-species level (subtilis-coli).
There are other aims that we are still working on such as use of this nanotube to perform more complex task using some more complicated genetic circuits.

Project Details

Step 0

The first step is about proving what the authors already did, to make sure that we can obtain the same results.

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Step 0.1

Antibiotic: It has been shown that the nanotube allow for a transient antibiotic resistance. In order to prove this part, we use two different strains of subtilis. Each have a different antibiotic resistance encoded and when grown together on a double selective medium colonies still form.

Step 0.2

GFP

Results