Difference between revisions of "Part:BBa K549001"

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[[Image:mikroskopgfp.png]] [[Image:mikroskopdurchlicht.png]]
 
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Transformed and induced ''E. coli'' cells, cultivated in LB-media (containing Ni(II)). Left-hand side: GFP-excitation; right-hand side: bright-field
  
  

Revision as of 15:38, 30 September 2011

Ni(II) dependent rcnA-promoter and GFP

E. coli has an RcnR-regulator that represses the rcnA-promoter in the absence of Ni(II). After addition of Ni (II) (NiCl2), RcnR binds this ion and is released from the promoter. Therefore, GFP will be expressed.

E. coli has to be used as reporter organism since RcnR is not coded on the plasmid.

In presence of nickel the Luciferase is expressed.

Usage and Biology

This BioBrick can be used to detect Ni(II)-Ions in water.

This is how it works:

Bild-rcnAGFP.png

The sensor is also sensing Co 2+, but not very sensitivly.

Experience: works

Mikroskopgfp.png Mikroskopdurchlicht.png Transformed and induced E. coli cells, cultivated in LB-media (containing Ni(II)). Left-hand side: GFP-excitation; right-hand side: bright-field


Sources: - [http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2958.2006.05369.x/pdf Nickel homeostasis in Escherichia coli – the rcnR-rcnA efflux pathway and its linkage to NikR function] Iwig JS, Rowe JL, Chivers PT
- [http://www.ncbi.nlm.nih.gov/pubmed?term=coordinating%20intracellular%20chivers Coordinating intracellular nickel-metal-site structure-function relationships and the NikR and RcnR repressors] Iwig JS, Chivers PT

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 863