Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jennifer Lei
Date created: 2007-10-24 11:00:00
Date modified: 2015-08-31 04:08:07
cmv-mLexa-dnaEN-Shp2-SV40PA
| Types | DnaRegion |
| Roles | Signalling engineered_region |
| Sequences | BBa_I756030_sequence (Version 1) |
Description
cmv-mLexA-dnaEN-Shp2-SV40PAWhen 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.
Notes
noneSource
EcoR1,Xba1,SacII (1-18bp)cmv from pcDNA3.1 zeo(+)(19-550bp)
kozak (551-556bp)
mLexA from pEG202 (557-1246bp)
dnEN from dnaE gene of Synechocystis sp.(split inteins)(1247-1630bp)
bsteII, Fse1, Kpn1 (1631-1657bp)
Shp2 from cDNA clone (clone ID #6409640) (1658-3439bp)
Stop Stop (3440-3445bp)
SV40 polyA (3446-3576bp)
asc1, spe1 (3577-3590bp)