Team:ETH Zurich/Biology/Cloning

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Revision as of 21:15, 10 September 2011

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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 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

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.

Design of our testsystem.

Characterization of LacIM1

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