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Team:Paris Bettencourt/Project
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| - | It has been shown that the nanotubes allow for a transient antibiotic resistance. In order to prove this part, we use two different strains of ''subtilis''. Each strain has a different antibiotic resistance encoded and when grown together on a double selective medium colonies still form. The limitations of this simple method is that we don't know if both strains survive or only one does. Different hypothesis follow from there: either both survive because both antibiotics are exchanged or one strain only survives because one antibiotic preferentially get through the | + | It has been shown that the nanotubes allow for a transient antibiotic resistance. In order to prove this part, we use two different strains of ''subtilis''. Each strain has a different antibiotic resistance encoded and when grown together on a double selective medium colonies still form. The limitations of this simple method is that we don't know if both strains survive or only one does. Different hypothesis follow from there: either both survive because both antibiotics are exchanged or one strain only survives because one antibiotic preferentially get through the nanotubes. |
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LacI, GFP | LacI, GFP | ||
Revision as of 14:04, 1 July 2011
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Contents |
Overall project
We started to work on one of the most intriguing microbiology discovery of the last decade: 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] in the Journal Cell 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 using synthetic biology to get a definite proof of the existence of nanotubes.
Our second aim is to caracterize the nanotubes: what passes through them and what are the typical diffusion times through the network. We will examine if RNA, proteins of different sizes and/or 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 that 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). As a general outline we will first investigate the inter-species (subtilis-coli) conneciton thanks to all the existing biobricks for E .coli then we will move on to an intra-species (subtilis to subtilis) connection and develop new parts specific to subtilis.
There are other aims that we are still working on such as use of this nanotube to perform more complex task (pattern formation for instance) using some more complicated genetic circuits.
Project Details
Step 0
The first step is about reproducing what the authors of the original paper already did, to make sure that we can obtain the same results.
Step 0.1
Antibiotic: It has been shown that the nanotubes allow for a transient antibiotic resistance. In order to prove this part, we use two different strains of subtilis. Each strain has a different antibiotic resistance encoded and when grown together on a double selective medium colonies still form. The limitations of this simple method is that we don't know if both strains survive or only one does. Different hypothesis follow from there: either both survive because both antibiotics are exchanged or one strain only survives because one antibiotic preferentially get through the nanotubes.
Step 0.2
LacI, GFP
