Signalling

Part:BBa_I756030

Designed by: Jennifer Lei   Group: iGEM07_Bay_Area_RSI   (2007-10-25)


cmv-mLexa-dnaEN-Shp2-SV40PA

cmv-mLexA-dnaEN-Shp2-SV40PA

When the SCFV is activated by contact with exposed MHC on cardiomyocytes that have been damaged in an infarction, the receptors will dimerise via interactions of the EporD2 domains. EporTransmembrane mutant version was used, L241G and L242P, to introduce a kink into the leucine zipper and to decrease the interchain interaction of EpoR for a tight on/off regulation of the receptor. Leucine zipper formation induces alpha helical structures that induce self-assembly by close packing of TM structures. Oligermerisation of wild-type EporTM is considerably high, producing a high basal rate of signaling which was not desired.

Upon dimerisation of the receptors, the intracellular portion of the chimeric receptor, GP130i (intracellular)will activate downstream signaling. JAK, a tryosine kinase, phosphorylates the gp130i which then Shp2 (a Src homology 2-containing tyrosine phosphatase) binds to. In our novel signaling system we have designed a version of Shp2 fused to dnaEN-mLexA. DnaEN and dnaEC are part of the split intein system which are designed to reconsitute mLexA-VP16 in vivo. MLexA-VP16 translocates to nucleus where is functions as the switch to upregulate expression of the rescue circuit, death circuit and cell adhesion circuit via the binding to the lexA operator minimal promoter.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1163
    Illegal XbaI site found at 7
    Illegal SpeI site found at 3585
    Illegal PstI site found at 1207
    Illegal PstI site found at 3031
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1163
    Illegal SpeI site found at 3585
    Illegal PstI site found at 1207
    Illegal PstI site found at 3031
    Illegal NotI site found at 15
    Illegal NotI site found at 1189
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1163
    Illegal BglII site found at 1093
    Illegal BglII site found at 2230
    Illegal BamHI site found at 1173
    Illegal XhoI site found at 1196
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1163
    Illegal XbaI site found at 7
    Illegal SpeI site found at 3585
    Illegal PstI site found at 1207
    Illegal PstI site found at 3031
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1
    Illegal EcoRI site found at 1163
    Illegal XbaI site found at 7
    Illegal SpeI site found at 3585
    Illegal PstI site found at 1207
    Illegal PstI site found at 3031
    Illegal NgoMIV site found at 1643
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3150
    Illegal BsaI.rc site found at 2369
    Illegal SapI.rc site found at 3248


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Parameters
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