Team:Paris Bettencourt/Experiments/YFP TetR diffusion experiments
From 2011.igem.org
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<h2>Conclusions</h2> | <h2>Conclusions</h2> | ||
- | <p>We investigated the presence of nanotubes by exposing our receiver TetO array in <i>B.subtilis</i> to the YFP:TetR fusion protein emitter, both in <i>E.coli</i> and <i>B.subtilis</i>. In both case, there was no appearance of bright fluorescence spots as expected with our characterization of the TetO array.</p> | + | <p>We investigated the presence of nanotubes by exposing our receiver TetO array in <i>B.subtilis</i> to the YFP:TetR fusion protein emitter, both in <i>E.coli</i> and <i>B.subtilis</i>. In both case, there was no appearance of bright fluorescence spots as expected with our characterization of the TetO array. So like in <em>T7 RNA polymerase diffusion experiments</em> we have no evidences of nanotube existence</p> |
Revision as of 02:57, 29 October 2011
![](https://static.igem.org/mediawiki/2011/1/19/Tubeornottube.png)
Testing nanotubes with the YFP concentrator system
Summary
All our experiments followed our microscopy protocol when not specified otherwise.
Results for the YFP concentrator:
- We've done E.coli to B.subtilis diffusion experiments (with negative results)
- We've done B.subtilis to B.subtilis diffusion experiments (with negative results)
Design overview
![](https://static.igem.org/mediawiki/2011/9/93/YFP_concentration_principle.jpg)
YFP:TetR/TetO array system
More information on the design here.
Emitter & receiver constructs in B.subtilis (receiver in plasmid)
Emitter construct in E.coli - Receiver construct in B.subtilis (plasmid)
We succeeded to integrate the TetO Array in B. subtilis with the multihost episomal pHM3 plasmid.
Experiments of E. coli (pFX234 plasmid, D. Lane strains) to B. subtilis (TetO Array, pHM3) to follow an eventual fluorescence diffusion by nanotube has been tried.
Movie : Experiment of mixed YFP:tetR (E. coli) & TetO Array (B. subtilis)
We cannot detect any tetR-YFP foci in the receiver cells specific of YFP concentration on the TetO Array. This suggests that the YFP-tetR fusion protein from E. coli cannot diffuse into B. subtilis through the potential nanotubes.
Conclusions
We investigated the presence of nanotubes by exposing our receiver TetO array in B.subtilis to the YFP:TetR fusion protein emitter, both in E.coli and B.subtilis. In both case, there was no appearance of bright fluorescence spots as expected with our characterization of the TetO array. So like in T7 RNA polymerase diffusion experiments we have no evidences of nanotube existence