Difference between revisions of "Part:BBa K1616019"
 
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The histidine kinase YF1 employs a light oxygen voltage blue light photosensor domain. In the absence of blue light, YF1 phosphorylate the regulator FixJ which induces gene expression from the FixK2 promoter. The opposite happens with pDawn.
 
The histidine kinase YF1 employs a light oxygen voltage blue light photosensor domain. In the absence of blue light, YF1 phosphorylate the regulator FixJ which induces gene expression from the FixK2 promoter. The opposite happens with pDawn.
  
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<center> [[Image:PDawnsystem.png|600px]] </center>
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<center> [[Image:PDawnsystem.png|800px]] </center>
  
  
 
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===Sequence and Features===
 
<partinfo>BBa_K1616019 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1616019 SequenceAndFeatures</partinfo>
  

Latest revision as of 15:09, 23 September 2015

pDawn system

The plasmids pDawn for light-activated gene expression. Basically, the YF1/FixJ drives gene expression from the pFixK2 promoter in a blue-light repressed manner. When we insert the λ phage repressor cI and the λ promoter pR in pDawn, this will invert the signal polarity and lead to the gene expression. The histidine kinase YF1 employs a light oxygen voltage blue light photosensor domain. In the absence of blue light, YF1 phosphorylate the regulator FixJ which induces gene expression from the FixK2 promoter. The opposite happens with pDawn.

PDawnsystem.png




Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 26
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1376
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1310
    Illegal NgoMIV site found at 1408
    Illegal NgoMIV site found at 2291
    Illegal NgoMIV site found at 2363
    Illegal NgoMIV site found at 2453
    Illegal NgoMIV site found at 2471
    Illegal NgoMIV site found at 2965
    Illegal AgeI site found at 2005
    Illegal AgeI site found at 3133
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3022
    Illegal BsaI.rc site found at 1904


Design Notes

The original sequence of pDawn have some illegal sites that have been removed.

Penn iGEM 2015 characterized the pDawn system as a light-sensitive receiver in its light-mediated cell communication system. With mRFP cloned downstream of the light-responsive promoter, the system's light-mediated gene expression was confirmed. Furthermore, the system was activated using small amount of artificial light produced with an LED. These small amounts reflect the intensity of light output from luminescent sender cells. For additonal information, follow this link: http://2015.igem.org/Team:Penn/Receiver.

Source

The pDawn & pDusk system, have been optimized and constructed in the way that the toxins sequences we introduce in our construction are in fact the reverse sequences.

References

Ohlendorf, Robert, Roee R. Vidavski, Avigdor Eldar, Keith Moffat, and Andreas Möglich. "From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression." Journal of Molecular Biology 416.4 (2012): 534-42. Web.