Team:Grenoble/Project/Main Results
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<h1>And after?</h1> | <h1>And after?</h1> | ||
- | In the future weeks (according to ours results) we will try to make the culture of our transformed bacteria in a medium | + | In the future weeks (according to ours results) we will try to make the culture of our transformed bacteria in a medium complemented with glucose and acetate. |
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+ | As we have had issues constructing paraBAD-gfp-<i>cyaA</i> using the standard restriction/ligation procedure, we are going to try another approach. We want to insert it on pSB3C5 (we have the brick but we are failing the insertion).</br> | ||
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A possible track would be to add another regulation system on the adenylate cyclase production with the riboswitch RsmA/rsmY. | A possible track would be to add another regulation system on the adenylate cyclase production with the riboswitch RsmA/rsmY. | ||
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Revision as of 02:11, 27 September 2012
Main results
This part will present the pre-regional Jamboree main results.Simple amplification loop
In a first place, biologists and modellers worked on a simple amplification loop.The modellers proved that a simple amplification loop could not work. There is no state where GFP is not expressed.
Why an AND gate ?
The team thought that an AND gate could be our solution. It could add a biological noise filter and resolve the problem of the continuous expression of GFP.Modelling results confirmed that built system will work.
Biological results
Biological AND gate test.Issues encountered
We have not yet constructed that part. We had some problem with the ligation procedure. Another explanation could be that our bacteria are transformed with the right construction leading to an over-expression of Adenylate cyclase which is lethal: this can be attributed to the fact that overproduction of cAMP is lethal to Escherichia coli possibly due to an accumulation of methylglyoxal [1][2].And after?
In the future weeks (according to ours results) we will try to make the culture of our transformed bacteria in a medium complemented with glucose and acetate.As we have had issues constructing paraBAD-gfp-cyaA using the standard restriction/ligation procedure, we are going to try another approach. We want to insert it on pSB3C5 (we have the brick but we are failing the insertion). A possible track would be to add another regulation system on the adenylate cyclase production with the riboswitch RsmA/rsmY.
References
- [1] Accumulation of Toxic Concentrations of Methylglyoxal by Wild-Type Escherichia coli K-12, Rollie S. Ackerman, Nicholas R. Cozzarelli and Wolfgang Epstein
- [2] Metabolic flux analysis of Escherichia coli in glucose-limited continuous culture. II. Dynamic response to famine and feast, activation of the methylglyoxal pathway and oscillatory behaviour
Jan Weber†, Anke Kayser‡ and Ursula Rinas