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! colspan="2" style="background:#FFBF00;"|PhoCl, a Mammalian Photocleavable Protein
 
! colspan="2" style="background:#FFBF00;"|PhoCl, a Mammalian Photocleavable Protein

Revision as of 22:56, 22 October 2017

E-cadherin_PhoCl Fusion Protein, a photosensitive cell adhesion protein

PhoCl, a Mammalian Photocleavable Protein
Function Photocleavable Linker
Use in Mammalian cells
Abstraction Hierarchy Part
RFC standard RFC10, RFC23 & RFC25 compatible
Backbone pSB1C3
Submitted by [http://2017.igem.org/Team:UCL UCL iGEM 2017]

As part of the UCL 2017's project "Light-induced Technologies" we investigated light sensitive proteins and their possible applications in synthetic genetic circuits. PhoCl is a novel (April 2017) photocleavable protein engineered from a green-to-red photoconvertible fluorescent protein.



Usage and Biology

As stated in the original paper: "The photoconversion reaction is a violet light (~400 nm)-induced β-elimination reaction that extends the conjugated system of the chromophore with concomitant cleavage of the polypeptide backbone to form an ~66-residue N-terminal fragment and an ~166-residue C-terminal fragment that remain associated."

The original protein has been engineered by Zhang et al. 2017 (Robert E. Campbell lab). The Campbell lab has sent the original plasmid to the UCL iGEM 2017 team as part of a collaboration and the team has made a BioBrick out of the engineered protein.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2550
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 752
    Illegal BamHI site found at 828
    Illegal BamHI site found at 944
    Illegal BamHI site found at 1868
    Illegal BamHI site found at 2170
    Illegal XhoI site found at 1552
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 208
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 65
    Illegal BsaI site found at 465
    Illegal BsaI site found at 872
    Illegal BsaI site found at 1414
    Illegal BsaI.rc site found at 306
    Illegal BsaI.rc site found at 3183



Functional Parameters


Reference

Zhang W, Lohman AW, Zhuravlova Y, Lu X, Wiens MD, Hoi H, Yaganoglu S, Mohr MA, Kitova EN, Klassen JS, Pantazis P, Thompson RJ, Campbell RE. Optogenetic control with a photocleavable protein, PhoCl. Nat Methods. 14(4):391-394 (2017) NCBI