Difference between revisions of "Part:BBa K1616022"
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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.
 
<center>https://static.igem.org/mediawiki/2015/9/9d/Pdawndusk.png</center>
 
<center>https://static.igem.org/mediawiki/2015/9/9d/Pdawndusk.png</center>
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H/E Holin and endolysin are toxins; after a period of late-gene expression, holin, an inner permeabilizing protein creates holes in the membrane in order to permeabilize it and then endolysin, a muralytic enzyme traverses the holes creating by holin and degraded the wall cell. This complex induce cell lysis of bacteria. (Young & Bläsi, 1995)
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Toxins will be produced after light stimulation and induce lysis.  
 
Toxins will be produced after light stimulation and induce lysis.  
  
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===References===
 
===References===
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Young, R., & Bläsi, U. (1995). Holins: Form and function in bacteriophage lysis. In FEMS Microbiology Reviews (Vol. 17, pp. 191–205). http://doi.org/10.1016/01686445(94)00079-4

Revision as of 14:00, 18 September 2015

pDawn - Endolysin/Holin


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.

Pdawndusk.png


H/E Holin and endolysin are toxins; after a period of late-gene expression, holin, an inner permeabilizing protein creates holes in the membrane in order to permeabilize it and then endolysin, a muralytic enzyme traverses the holes creating by holin and degraded the wall cell. This complex induce cell lysis of bacteria. (Young & Bläsi, 1995)
Toxins will be produced after light stimulation and induce lysis.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1405
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2755
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2689
    Illegal NgoMIV site found at 2787
    Illegal NgoMIV site found at 3670
    Illegal NgoMIV site found at 3742
    Illegal NgoMIV site found at 3832
    Illegal NgoMIV site found at 3850
    Illegal NgoMIV site found at 4344
    Illegal AgeI site found at 306
    Illegal AgeI site found at 376
    Illegal AgeI site found at 3384
    Illegal AgeI site found at 4512
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 4401
    Illegal BsaI.rc site found at 3283




Design Notes

We obtained the plasmids from the Centre for Biological Signalling Studies and the University of Freiburg. As soon as we received them, we observed that the multiple cloning site as well as the λ promoter pR were located upstream the YF1/FixY system.

Source

This part is composed of Holin/Endolysin toxin BBa_K1616005 and pDawn system BBa_K1616019

References

Young, R., & Bläsi, U. (1995). Holins: Form and function in bacteriophage lysis. In FEMS Microbiology Reviews (Vol. 17, pp. 191–205). http://doi.org/10.1016/01686445(94)00079-4