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Team:Paris Bettencourt/Experiments/YFP TetR diffusion
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<p>The planning of the experiments is the following : first we have tested the strains from D.Lane containing YFP:tetR and tetO array. Then we constructed/biobricked the YFP:tetR and tetO array system. To finish with the microscopy step and results of this proof of concept between <i>B. subtilis and B. subtilis / E. coli</i>.</p> | <p>The planning of the experiments is the following : first we have tested the strains from D.Lane containing YFP:tetR and tetO array. Then we constructed/biobricked the YFP:tetR and tetO array system. To finish with the microscopy step and results of this proof of concept between <i>B. subtilis and B. subtilis / E. coli</i>.</p> | ||
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<h2>Testing the YFP:tetR strains from D. Lane</h2> | <h2>Testing the YFP:tetR strains from D. Lane</h2> | ||
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| + | In the article <a href="http://onlinelibrary.wiley.com/doi/10.1046/j.1365-2958.2003.03837.x/pdf">[1]</a>, strains are growing at 20°C to avoid protein agregation but the problem is that nanotube between <i>B. subtilis</i> has been only proved to exist at 37°C. | ||
| + | We test different possibility : at 37°C or 30°C and concentration of arabinose (0% - 0,1% -0,2%) | ||
<h2>Biobricked system construction</h2> | <h2>Biobricked system construction</h2> | ||
| - | <h2> | + | <h2>Results and microscopy of the proof of concept</h2> |
<p><em>In the emittor cell <i>(B. Subtilis)</i></em>, we have inserted a expressive system for the YFP:tetR. It contains the promoter pVeg, the RBS for B. Subtilis and the YFP:tetR protein. Production of YFP:tetR will diffuse throught the nanotube to the receiver cell.</p> | <p><em>In the emittor cell <i>(B. Subtilis)</i></em>, we have inserted a expressive system for the YFP:tetR. It contains the promoter pVeg, the RBS for B. Subtilis and the YFP:tetR protein. Production of YFP:tetR will diffuse throught the nanotube to the receiver cell.</p> | ||
Revision as of 14:02, 12 September 2011
Experiments of the YFP concentration design
The planning of the experiments is the following : first we have tested the strains from D.Lane containing YFP:tetR and tetO array. Then we constructed/biobricked the YFP:tetR and tetO array system. To finish with the microscopy step and results of this proof of concept between B. subtilis and B. subtilis / E. coli.
Testing the YFP:tetR strains from D. Lane
In the article [1], strains are growing at 20°C to avoid protein agregation but the problem is that nanotube between B. subtilis has been only proved to exist at 37°C. We test different possibility : at 37°C or 30°C and concentration of arabinose (0% - 0,1% -0,2%)Biobricked system construction
Results and microscopy of the proof of concept
In the emittor cell (B. Subtilis), we have inserted a expressive system for the YFP:tetR. It contains the promoter pVeg, the RBS for B. Subtilis and the YFP:tetR protein. Production of YFP:tetR will diffuse throught the nanotube to the receiver cell.
In the receiver cell (B. Subtilis or E. Coli), there is the tetO array where diffused YFP:tetR will concentrate. The YFP is the monitor of the signal.
The principle of the design is summed up in the image below
Fig1: Schematics of the YFP concentration design
