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Team:Paris Bettencourt/Experiment/T7 diff subt subt microfluidic
From 2011.igem.org
Nanotube-assisted diffusion of T7 polymerase in Microfluidics
Experimental Scheme
In order to test whether our Bacillus subtilis T7 emitters & receivers can form nanotubes when mixed, we mix them in microfluidic system modified from Jeff Hasty's recent paper [1], shown below. For more detailed information regarding the microfluidic device and experimental procedure, check our methodology page.
We imaged two channels: one experimental channel injected with an emitter strain (RFP constitutive) and a receiver strain (pT7-T7polyermase-GFP); the other is a control channel injected with only the receiver strain. The receiver strain contains the T7 autoloop, which will gain a strong fluorescence when activated.
Results
The receiver strain by itself in our control channel have never produced any GFP positive cells, in contrast to the leaky situation where the T7 autoloop is encoded by a plasmid in E.coli. This is due to the low copy number of T7 polymerase gene on the chromosome, d
References
- Entrainment of a population of synthetic genetic oscillators. Mondragón-Palomino, O., Danino, T., Selimkhanov, J., Tsimring, L. & Hasty, J. Science 333, 1315-1319 (2011).
