Difference between revisions of "Part:BBa K4375004"

 
Line 5: Line 5:
 
BLADE is a modified AraC transcription factor engineered to react to blue light instead of arabinose. Thus its arabinose binding and homodimerising domain were swapped to VVD.   
 
BLADE is a modified AraC transcription factor engineered to react to blue light instead of arabinose. Thus its arabinose binding and homodimerising domain were swapped to VVD.   
  
−
<!-- Add more about the biology of this part here
+
 
−
===Usage and Biology===
+
==Usage and Biology==
 +
 
 +
BLADE protein is produced from AraC by exchanging the dimerization domain of AraC with a light-inducible dimerization domain. The switching of this engineered AraC from monomer to dimer is controlled with light.  In its monomeric form, the engineered AraC would contact the high-affinity I1 half-site16, but not the low-affinity I2 half-site, needed to recruit the RNA polymerase. For the light-triggered dimerization domain Vivid (VVD) domain was selected, which has often been successfully used to control, with light, the dimerization of proteins of interest. VVD senses blue light via the flavin adenine dinucleotide (FAD) chromophore. Blue light triggers the formation of a cysteinyl-flavin adduct, which generates a new hydrogen-bond network that releases the N terminus (N-terminal cap) from the protein core and restructures it, creating a new dimerization interface.
 +
 
  
 
<!-- -->
 
<!-- -->
Line 12: Line 15:
 
<partinfo>BBa_K4375004 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4375004 SequenceAndFeatures</partinfo>
  
 +
===References===
 +
 +
https://www.nature.com/articles/s41589-021-00787-6.pdf
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Revision as of 09:02, 7 October 2022


BLADE (light-responsive AraC transcription factor)

BLADE is a modified AraC transcription factor engineered to react to blue light instead of arabinose. Thus its arabinose binding and homodimerising domain were swapped to VVD.


Usage and Biology

BLADE protein is produced from AraC by exchanging the dimerization domain of AraC with a light-inducible dimerization domain. The switching of this engineered AraC from monomer to dimer is controlled with light. In its monomeric form, the engineered AraC would contact the high-affinity I1 half-site16, but not the low-affinity I2 half-site, needed to recruit the RNA polymerase. For the light-triggered dimerization domain Vivid (VVD) domain was selected, which has often been successfully used to control, with light, the dimerization of proteins of interest. VVD senses blue light via the flavin adenine dinucleotide (FAD) chromophore. Blue light triggers the formation of a cysteinyl-flavin adduct, which generates a new hydrogen-bond network that releases the N terminus (N-terminal cap) from the protein core and restructures it, creating a new dimerization interface.


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
    COMPATIBLE WITH RFC[1000]

References

https://www.nature.com/articles/s41589-021-00787-6.pdf