Team:ESBS-Strasbourg/Results/Characterization
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- | This led us to make two hypotheses:<br> | + | This led us to make two hypotheses:<br><br> |
- | -Either the fluorescence efficiency fusion protein had a better quantum yield due to a better folding/environment around its fluorophore.<br> | + | -Either the fluorescence efficiency fusion protein had a better quantum yield due to a better folding/environment around its fluorophore.<br><br> |
- | -Or, most probably, PIF6-linker-GFP expression is stronger than GFP expression, due to the PIF6-linker sequence in Nter of GFP<br> | + | -Or, most probably, PIF6-linker-GFP expression is stronger than GFP expression, due to the PIF6-linker sequence in Nter of GFP.<br> |
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- | In order to find an answer to this question, we calculated specific fluorescence by measuring both fluorescence and amount of fluorescent protein. First we measured fluorescences of three different E.coli lysats from one GFP negative strain, one GFP expressing strain and one PIF6-linker-GFP expressing strain. | + | In order to find an answer to this question, we calculated specific fluorescence by measuring both fluorescence and amount of fluorescent protein. First we measured fluorescences of three different E.coli lysats from one GFP negative strain, one GFP expressing strain and one PIF6-linker-GFP expressing strain. The table below shows the tests made on the previews tubes. Cells were resuspended and sonicated and the lysate was analyzed with a fluorimetre (absorption wavelenght=490nm, emission wavelenght=510nm). The test was done 3 times to give an average measurement. We then calculated the specific fluorescence according to the OD600, the results are also shown in the table.<br> |
- | The table below shows the tests made on the previews tubes. Cells were resuspended and sonicated and the lysate was analyzed with a fluorimetre (absorption wavelenght= 490nm, emission wavelenght= 510nm). The test was done 3 times to give an average measurement. We then calculated the specific fluorescence according to the OD600, the results are also shown in the table.<br> | + | |
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Revision as of 22:29, 27 October 2010
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