Difference between revisions of "Part:BBa K549001"

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<partinfo>BBa_K549001 short</partinfo>
 
<partinfo>BBa_K549001 short</partinfo>
  
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E. coli has an RcnR-regulator that switches on the RcnA-Promotor in the presence of Ni(II). After induction GFP will be expressed.
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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.
 
E. coli has to be used as reporter organism since RcnR is not coded on the plasmid.
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In presence of nickel the Luciferase is expressed.
  
 
===Usage and Biology===
 
===Usage and Biology===
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The sensor is also sensing Co 2+, but not very sensitivly.
 
The sensor is also sensing Co 2+, but not very sensitivly.
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The promoter sequence of pnikA is missing in the part sequence below. This is the promoter sequence of pnikA directly upstream of the reporter gene. The promoter sequence also contains the first ca. 50 nt of nikA, i.e. its a translational fusion.
 
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ACGGATTGTATGAGACATGGCAACACCTGGTTAACAAGAATATGAAAAATCATAGCACTATTAATCTACTGGGGGGTAGT
 
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ATCAGGTACTGGGGGGGAGTAGAATCAGATTGCCGAATTAATACTAAGAATTATTATCATGACCGAATTTACAACTCTTC
 
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TTCAGCAAGGAAACGCCTGGTTCTTCATCCCCAGCGCCATCTTACTTGGTGCG
 
  
  

Latest revision as of 17:34, 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.


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