Composite

Part:BBa_K1936003

Designed by: Dorian Röders   Group: iGEM16_Duesseldorf   (2016-10-13)


PhyB-Vp16


Usage and Biology

Phytochrome B (PhyB) is a photoreceptor from Arabidopsis thaliana. The interacting factor of PhyB is PIF6 which can be bound after a light induced conformational change of PhyB. By the absorption of red light PhyB is activated (binds to PIF6). PhyB can also be inactivated (state in which it is not bound to PIF6) by far red light. To react to light PhyB requires the binding of the chromophore phycocyanobilin (PCB), which is not produced in mammalian and yeast cells and most bacteria.[1]

PhyB is linked to the herpes simplex VP16 transactivation domain. VP16 is a 65 kD popypeptide originally encoded in herpes simplex virus type 1 that primes transcription from the five virally encoded immediate early genes.[2] In scientific use it usually linked to tetR (tetracyclin Repressor) which binds a motif in the upstream region of a gene (tetO).[1] This brings VP16 close to the DNA where it can recruit RNA polymerase and therefore induces expression of the related gene.[3]

Biobrick in combination with tetR-PIF6:
In red light, PhyB-VP16 is recruited to PIF6 at the promoter site, which activates the expression of the genes. Upon absorption of far red light the interaction of Phyb and PIF6 is terminated, resulting in silencing of the gene. [1]


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1951
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1951
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1951
    Illegal BglII site found at 490
    Illegal BamHI site found at 572
    Illegal XhoI site found at 523
    Illegal XhoI site found at 542
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1951
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1951
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 739


References

[1] Konrad Müller, Matias D Zurbriggen and Wilfried Weber(2014) Control of gene expression using a red- and far-red light-responsive bi-stable toggle switch. Doi:10.1038/nprot.2014.038 February 2014
[2] Steven J. Triezenberg, Robert C. Kingsbury and Steven L. McKnight Functional dissection of VP16, the transactivator of herpes simplex virus immediate early gene expression. Cold Spring Harbor laboratory press
[3] Eloi Gari, Lidia Piedrafita, Marti Aldea and Enrique Herrero (1996) A Set of Vectors with a Tetracycline-Regulatable Promoter System for Modulated Gene Expression in Saccharomyces cerevisiae. Yeast vol. 13: 837-848 (1997)

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