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Team:INSA-Lyon/Project/Stage3/Results
From 2010.igem.org
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<h2>Results</h2> | <h2>Results</h2> | ||
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| - | <h3>Caracterisation Curli</h3> | + | <h3>Caracterisation Curli</h3><br> |
<p>We studied the Curli promoter with the PHL1273 chassis which contain Curli promoter and the mutation ompR234. This mutation is required for the high expression of the Curli promoter. A modified GFP is behind the promoter. This GFP can’t be catabolized by the usual enzyme, so in a period of 24 hours all the produced GFP remained. The fluorescence of the culture is proportionnal to the quantity of GFP and thus, to the activity of the Curli promoter. | <p>We studied the Curli promoter with the PHL1273 chassis which contain Curli promoter and the mutation ompR234. This mutation is required for the high expression of the Curli promoter. A modified GFP is behind the promoter. This GFP can’t be catabolized by the usual enzyme, so in a period of 24 hours all the produced GFP remained. The fluorescence of the culture is proportionnal to the quantity of GFP and thus, to the activity of the Curli promoter. | ||
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| - | + | <p>First of all the culture were in a waterbath at 28°C and only the shaking speed changed. We try 9 shaking speeds from 50 to 200 rpm.</p> | |
Revision as of 15:02, 26 October 2010
Results
Caracterisation Curli
We studied the Curli promoter with the PHL1273 chassis which contain Curli promoter and the mutation ompR234. This mutation is required for the high expression of the Curli promoter. A modified GFP is behind the promoter. This GFP can’t be catabolized by the usual enzyme, so in a period of 24 hours all the produced GFP remained. The fluorescence of the culture is proportionnal to the quantity of GFP and thus, to the activity of the Curli promoter. We chose to study the influence of shaking speed, temperature and osmotic pressure. For all the experiment, we used the same protocoles (Mesurement of fluo and ….). We were three operators to do the same experiment at the same time.
The culture of reference was a culture of PHL818, which also contains the mutation OmpR234. The Curli promoter and the GFP gene was introduced in this chassis in order to create PHL1273.
- Influence of the shaking speed
First of all the culture were in a waterbath at 28°C and only the shaking speed changed. We try 9 shaking speeds from 50 to 200 rpm.
