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Team:ETH Zurich/Biology/Cloning
From 2011.igem.org
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| + | == '''Used parts from registry''' == | ||
| + | * BBa R0040 P<sub>Tet</sub> | ||
| + | * BBa R0051 λ<sub>P</sub> | ||
| + | * BBa R0010 P<sub>lac</sub> | ||
| + | * BBa R0061 P<sub>lux</sub> | ||
| + | * BBa J23100 P<sub>const</sub> | ||
| + | * BBa C0061 LuxI<sub>LVA</sub> | ||
| + | * BBa C0040 TetR<sub>LVA</sub> | ||
| + | * BBa C0051 CI | ||
| + | * BBa C0062 LuxR | ||
| + | * BBa B0015 Double Terminator | ||
| + | |||
| + | |||
| + | == '''Synthesized parts''' == | ||
| + | * LacI<sub>M1</sub> | ||
| + | * AlcR (codon optimized) | ||
== '''Design of a AlcR/acetaldeyhde repressed promotor P<sub>AlcR</sub>''' == | == '''Design of a AlcR/acetaldeyhde repressed promotor P<sub>AlcR</sub>''' == | ||
Revision as of 08:10, 1 September 2011
Used parts from registry
- BBa R0040 PTet
- BBa R0051 λP
- BBa R0010 Plac
- BBa R0061 Plux
- BBa J23100 Pconst
- BBa C0061 LuxILVA
- BBa C0040 TetRLVA
- BBa C0051 CI
- BBa C0062 LuxR
- BBa B0015 Double Terminator
Synthesized parts
- LacIM1
- AlcR (codon optimized)
Design of a AlcR/acetaldeyhde repressed promotor PAlcR
For the promotor PAlcR the operator site of AlcR-inducer from apergillus nidulans was placed between the -10 and -35 region of the strong promotor. While acetaldeyhde is bind to AlcR, the complex will bind to the operon and block the binding of the RNA-polymerase.
Design of the test-system
To test our designed AlcR-promotors the following system was designed (see Figure). To induce the expression of AlcR a tetracycline induced promotor was used. To monitor the expression of AlcR a His-tag was introduced. In present of acetaldeyhde AlcR binds to the AlcR operon and inhibits the expression of GFP. For a better signal assam gfp was used.
