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Team:ETH Zurich/Biology/Cloning
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| - | [[File:AlcR_Promotordesign.png|600px|center|thumb|'''Design of the promotor for the AlcR-system''', the operon sequence is designed between the -10 and the -35 region (blue) of a strong promotor.]] | + | |
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| + | == '''Design of a AlcR/acetaldeyhde repressed promotor P<sub>AlcR</sub>''' == | ||
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| + | For the promotor P<sub>AlcR</sub> 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. | ||
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| + | [[File:AlcR_Promotordesign.png|600px|center|thumb|'''Design of the promotor for the AlcR-system''', the operon sequence is designed between the -10 and the -35 region (blue) of a strong promotor with inverted and direct-repeats.]] | ||
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| + | == '''Design of the test-system''' == | ||
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| + | 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. | ||
| + | [[File:Testsystem.png|600px|center|thumb|'''Design of our testsystem'''.]] | ||
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| + | == '''Design of the parts''' == | ||
Revision as of 08:39, 25 August 2011
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.
