Team:ESBS-Strasbourg/Results/Characterization

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Revision as of 22:09, 27 October 2010

ESBS - Strasbourg

	Let me guide you	PIF6-linker-GFP In the course of our experiments, we were led to express PIF-linker-GFP to check if this fusion protein was fluorescent before going into further characterization of our system. Accordingly to the green fluorescence of the strain, we successfully expressed PIF6-linker-GFP under constitutive (promotor J23100 and RBS B0034) control (PIF3-linker-GFP is still in progress). This means this upstream tagging sequence does not prevent translation of the downstream protein. One really surprising result yet was that colonies expressing PIF6-linker-GFP were from far more fluorescent than those expressing GFP only. This came to a surprise since the expression of both proteins are controlled by the same promoter/RBS and that GFP-fusion proteins are more likely to be less fluorescent than GFP alone! Figure 1 : Picture of the pellets of the different strains. On the left is the negative control with no GFP within the E. coli cells. In the middle is the positive control with a culture of E. coli expressing the GFP. On the right is the culture of E. coli expressing PIF6-linker-GFP. It is possible to clearly see the difference in fluorescence with a PIF6-linker-GFP pellet greener than the two others. b

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Retrieved from "http://2010.igem.org/Team:ESBS-Strasbourg/Results/Characterization"

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-In the course of our experiments, we were led to express PIF-linker-GFP to check if this fusion protein was fluorescent before going into further characterization of our system.<br>
+In the course of our experiments, we were led to express PIF-linker-GFP to check if this fusion protein was fluorescent before going into further characterization of our system. Accordingly to the green fluorescence of the strain, we successfully expressed PIF6-linker-GFP under constitutive (promotor J23100 and RBS B0034) control (PIF3-linker-GFP is still in progress). This means this upstream tagging sequence does not prevent translation of the downstream protein. <br>
-We successfully expressed PIF6-linker-GFP under constitutive (promotor J23100 and RBS B0034) control (PIF3-linker-GFP is still in progress). As expected, the assembly of the parts works as predicted and neither the PIF6 nor the linker interfere with the GFP. It shows that those two factors could be assembled with any protein that would be targeted for degradation.<br>
+One really surprising result yet was that colonies expressing PIF6-linker-GFP were from far more fluorescent than those expressing GFP only. This came to a surprise since the expression of both proteins are controlled by the same promoter/RBS and that GFP-fusion proteins are more likely to be less fluorescent than GFP alone!
-One really surprising result was that colonies expressing PIF6-linker-GFP were from far more fluorescent than those expressing GFP only. This came to a surprise since the expression of both proteins are controlled by the same promoter/RBS and that GFP-fusion proteins are more likely to be less fluorescent than GFP alone!
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+Figure 1 : Picture of the pellets of the different strains. On the left is the negative control with no GFP within the E. coli cells. In the middle is the positive control with a culture of E. coli expressing the GFP. On the right is the culture of E. coli expressing PIF6-linker-GFP. It is possible to clearly see the difference in fluorescence with a PIF6-linker-GFP pellet greener than the two others.
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