Coding

Part:BBa_K1720000:Design

Designed by: YuanbinCui   Group: iGEM15_SCUT-China   (2015-08-09)
Revision as of 10:44, 9 August 2015 by Registry (Talk | contribs)

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Human guanylate cyclase1,soluble, alpha 3 unit


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1498
    Illegal PstI site found at 1827
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1498
    Illegal PstI site found at 1827
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1498
    Illegal PstI site found at 1827
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1498
    Illegal PstI site found at 1827
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1576
    Illegal SapI site found at 1446


Design Notes

This subunit coding part with a GFP reporter was then transfected into HEK293 cells and in vivo green fluorescence signal was observed under fluorescence microscope. After that we used lentiviral vector to transfected alpha 3 unit and beta 3 unit to HEK293 cells together. Since soluble guanylate cyclases (sGC) are heterodimeric proteins that catalyze the conversion of GTP to 3',5'-cyclic GMP(cGMP) and pyrophosphate. The level of cGMP will be up regulated.The positive control was HEK293 cells that treat with Sodium Nitroprusside ,a NO donator that activate sGC and up regulate the level of cGMP. A negative control was made by transfecting an empty vector that does not contain sGC subunit. We used Elisa to detect cGMP level.


Source

http://www.ncbi.nlm.nih.gov/gene/2982

References