Team:ESBS-Strasbourg/Results/Characterization
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If we correlate the fluorescence with the optical density, we can obtain a difference fluorescence factor of: 15,47. It means that the PIF6-linker-GFP gives rise to 15,5 times more fluorescence per OD unit than the normal GFP. | If we correlate the fluorescence with the optical density, we can obtain a difference fluorescence factor of: 15,47. It means that the PIF6-linker-GFP gives rise to 15,5 times more fluorescence per OD unit than the normal GFP. | ||
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<imf src="https://static.igem.org/mediawiki/2010/4/43/ESBS-Strasbourg-Char1.png"> | <imf src="https://static.igem.org/mediawiki/2010/4/43/ESBS-Strasbourg-Char1.png"> | ||
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+ | Figure 2 : Relative specific fluorescence of GFP-, GFP and PIF6-linker-GFP expressing strains. | ||
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Simultaneously, we quantified the amount of overexpressed protein (either GFP or PIF6-linker-GFP) with a SDS-PAGE (see figure 2) | Simultaneously, we quantified the amount of overexpressed protein (either GFP or PIF6-linker-GFP) with a SDS-PAGE (see figure 2) | ||
The GFP strain showed a new (or stronger) spot at around 27 kDa compared with the GFP negative control, which perfectly fits GFP's molecular weight. While the expected molecular weight of PIF6-linker-GFP is around 43 kDa, the PIF6-linker-GFP strain showed at least 6 new spots when compared with the negative control, ranking from 43 to 32 kDa, which could be interpreted as truncated proteins. We thus performed an immunoblot assay using anti-GFP antibodies in order to see whether these spots were all responsible for the fluorescence within the PIF6-linker-GFP strain. | The GFP strain showed a new (or stronger) spot at around 27 kDa compared with the GFP negative control, which perfectly fits GFP's molecular weight. While the expected molecular weight of PIF6-linker-GFP is around 43 kDa, the PIF6-linker-GFP strain showed at least 6 new spots when compared with the negative control, ranking from 43 to 32 kDa, which could be interpreted as truncated proteins. We thus performed an immunoblot assay using anti-GFP antibodies in order to see whether these spots were all responsible for the fluorescence within the PIF6-linker-GFP strain. |
Revision as of 00:28, 28 October 2010
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