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Team:EPF-Lausanne/Our Project/Assembly/Plac
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We used a medium-strength Plac promoter that had been designed by Henrike Niederholtmeyer and put it into biobrick format. We characterized it by coupling the promoter to RFP and adding increasing concentrations of IPTG. | We used a medium-strength Plac promoter that had been designed by Henrike Niederholtmeyer and put it into biobrick format. We characterized it by coupling the promoter to RFP and adding increasing concentrations of IPTG. | ||
Revision as of 15:41, 20 September 2011
Plac promoter characterization
We used a medium-strength Plac promoter that had been designed by Henrike Niederholtmeyer and put it into biobrick format. We characterized it by coupling the promoter to RFP and adding increasing concentrations of IPTG.
We didn't transform a LacI gene in the DH5alpha cells. Still, these cells can have a basal expression of the transcription factor. By adding IPTG to the cell's medium, we make sure to inhibit any endogenous LacI expression, in order to have the maximal RFP expression that can be driven by our Plac biobrick.
IPTG induction
The DH5alphas cells, even if the plasmid we transform doesn't contain TetR, can still have a basal expression of the transcription factor. By adding ATC, we are able to inhibit this TetR basal expression, revealing the full power of Ptet as a promoter sequence.
Cells without IPTG: [[File:|460px]]
Cells with IPTG: [[File:|480px]]
Experimental results:
[[File:|600px]]
Dose-response curve
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