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Team:Duke/Project/Results/Part2
From 2013.igem.org
(Difference between revisions)
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TALEs | TALEs | ||
| - | Initial characterization: TALE-mCherry constructs are present and expression is inducible by beta-estradiol. | + | #Initial characterization: TALE-mCherry constructs are present and expression is inducible by beta-estradiol. Cells were grown with and without beta-estradiol and fluorescence was compared to background levels using a control strain. |
| - | + | ||
<br> | <br> | ||
insert graphs/data here | insert graphs/data here | ||
<br> | <br> | ||
| + | #Relationship between beta-estradiol concentration and TALE-mCherry expression. Cells were grown in varied beta-estradiol concentrations and fluorescence was compared to background levels using a control strain | ||
| + | <br> | ||
| + | insert input/output curve here | ||
| + | <br> | ||
| + | |||
ACT1 reporters | ACT1 reporters | ||
| - | Initial characterization: ACT1-yEVenus constructs | + | #Initial characterization: ACT1-yEVenus constructs with no operators showed distinct constitutive gene expression. However, presence of operator sites in the 5' UTR knocked down constitutive expression almost to background levels. In lieu of these results, we decided to construct TEF1-promoter reporter constructs. TEF1 is a stronger constitutive promoter that we hoped would counteract the effects of the operator sequence on constitutive expression levels. |
| + | <br> | ||
| + | insert graphs/data here | ||
| + | <br> | ||
| + | |||
| + | TEF1 reporters | ||
| + | |||
| + | #Initial characterization: TEF1-yEVenus constructs with operator sites in the 5' UTR showed expression levels significantly higher than background levels, and comparatively higher than the ACT1-yEVenus constructs. | ||
| + | <br> | ||
| + | insert graphs/data here | ||
| + | <br> | ||
Revision as of 06:15, 27 September 2013
Results: Part Characterization using Flow Cytometry
After being integrated into the yeast genome, parts were characterized individually by flow cytometry.
TALEs
- Initial characterization: TALE-mCherry constructs are present and expression is inducible by beta-estradiol. Cells were grown with and without beta-estradiol and fluorescence was compared to background levels using a control strain.
insert graphs/data here
- Relationship between beta-estradiol concentration and TALE-mCherry expression. Cells were grown in varied beta-estradiol concentrations and fluorescence was compared to background levels using a control strain
insert input/output curve here
ACT1 reporters
- Initial characterization: ACT1-yEVenus constructs with no operators showed distinct constitutive gene expression. However, presence of operator sites in the 5' UTR knocked down constitutive expression almost to background levels. In lieu of these results, we decided to construct TEF1-promoter reporter constructs. TEF1 is a stronger constitutive promoter that we hoped would counteract the effects of the operator sequence on constitutive expression levels.
insert graphs/data here
TEF1 reporters
- Initial characterization: TEF1-yEVenus constructs with operator sites in the 5' UTR showed expression levels significantly higher than background levels, and comparatively higher than the ACT1-yEVenus constructs.
insert graphs/data here
