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Team:Paris Bettencourt/Experiments/YFP TetR diffusion experiments
From 2011.igem.org
Testing nanotubes with the YFP concentrator system
Summary
Results for the YFP concentrator:
- We've done E.coli to B.subtilis diffusion experiments (with negative results)
- We successfully BioBricked both the YFP:TetR (BBa_K606025) and the TetO Array (BBa_K606026) constructs and sent them to the registry
- We characterized the YFP:TetR fusion protein both in E.coli and B.subtilis
- We characterized the TetO array both in E.coli and B.subtilis
Design overview
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 success 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.
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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.
