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Revision as of 08:03, 11 September 2011 (view source) Smurmann (Talk | contribs) (→Design of a AlcR/acetaldeyhde repressed promotor PAlcR) ← Older edit		Revision as of 08:04, 12 September 2011 (view source) Sabineoesterle (Talk | contribs) (→Design of a AlcR/acetaldehyde repressed promotor PAlcR) Newer edit →
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	* LacI<sub>M1</sub>		* LacI<sub>M1</sub>
	* AlcR (codon optimized)		* AlcR (codon optimized)
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-	== '''Design of a AlcR/acetaldehyde 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.]]
	== '''Design of the test-system''' ==		== '''Design of the test-system''' ==

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Revision as of 08:04, 12 September 2011

Used parts from registry

• BBa R0040 P_Tet
• BBa R0051 λ_P
• BBa R0010 P_lac
• BBa R0061 P_lux
• BBa J23100 P_const
• BBa C0061 LuxI_LVA
• BBa C0040 TetR_LVA
• BBa C0051 CI
• BBa C0062 LuxR
• BBa B0015 Double Terminator

Synthesized parts

• LacI_M1
• AlcR (codon optimized)

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.

Characterization of LacI_M1