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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- | <img src="https://static.igem.org/mediawiki/2010/4/43/ESBS-Strasbourg-Char1.png"> | + | <img src="https://static.igem.org/mediawiki/2010/4/43/ESBS-Strasbourg-Char1.png" width="70" height="85"> |
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Chart 1 : Relative specific fluorescence of GFP-, GFP and PIF6-linker-GFP expressing strains. | Chart 1 : Relative specific fluorescence of GFP-, GFP and PIF6-linker-GFP expressing strains. | ||
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- | Simultaneously, we | + | Simultaneously, we wanted to quantify the amount of overexpressed protein (either GFP or PIF6-linker-GFP) with a SDS-PAGE (see figure 2).<br> |
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. | ||
- | Result of this immunoblot is also reported in the figure 2. | + | Result of this immunoblot is also reported in the figure 2 below. |
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+ | <img src="https://static.igem.org/mediawiki/2010/9/9f/ESBS_Strasbourg_Figure_2.png" width="70" height="85"> | ||
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+ | Figure 2 : Pictures of the SDS gel (on the left), and the corresponding immunoblot (on the right). The immunoblot uses rabbit anti-GFP antibodies. | ||
+ | </center> | ||
Revision as of 00:53, 28 October 2010
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