Difference between revisions of "Part:BBa K4444002"
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===Usage and Biology===
 
===Usage and Biology===
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Our project builds on  the protein circuit RELEASE (Vlahos et al.) by introducing light-inducible  proteins or photoswitches. Photocleavable protein (PhoCl) is a recently developed optogenetic technology that self-cleaves  when stimulated by 405 nm ultraviolet light. PhoCl was engineered from a green-to-red photoconvertible fluorescent protein. When stimulated with ultraviolet light, the protein undergoes a β-elimination reaction, producing a small peptide fragment and a large barrel fragment that will spontaneously dissociate (Zhang et al.).
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Mechanism: Upon light-activation PhoCl breaks, removing the ER retention motif and allowing the protein to continue along the secretion pathway. The furin cut site will later be cleaved at the Golgi body, allowing the POI to be freely floating in the vesicle. At the plasma membrane the POI will be secreted.
  
 
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Revision as of 07:50, 12 October 2022

PhoCl Secrete GFP

PhoCl Secrete mRuby is modified from the RELEASE system. It contains a GFP as a protein of interest, a furin cut site, PhoCl2c, tri-transmembrane domain, and an ER retention motif. PhoCl Secrete will completely release a protein of interest in response to ultra violet light stimulation.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 52
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 775
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 946
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 60
    Illegal BsaI.rc site found at 1468