Difference between revisions of "Part:BBa K549003"
(→‎Usage and Biology)
 
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[[Image:Bild-rcnaluxAB.png]]
 
[[Image:Bild-rcnaluxAB.png]]
  
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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
 
  
 
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Latest revision as of 15:22, 30 September 2011

Nickel(II) dependent promotor with luxAB as reporter

The biobrick contains the RcnA promotor from E. coli fused with the luxAB reporter gene. E. coli strains own the RcnR-regulator gene which directly regulates the transcription of the RcnA nickel-cobalt efflux protein from which the promotor is amplified. In absence of Ni(II), RcnR represses the rcnA-promoter. After addition of Ni (II) (NiCl2), RcnR binds this ion and is released from the promoter. Therefore, luxAB will be expressed.

--> also use luxCDE or luciferine to detect output.

In presence of nickel the Luciferase is expressed.

The promoter is also a little bit sensitive for Co 2+.



Usage and Biology

This is how the BioBrick is working:

Bild-rcnaluxAB.png


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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 751
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1270