Regulatory

Part:BBa_K203107

Designed by: Corinna Hiller and Hannah Uckelmann   Group: iGEM09_Heidelberg   (2009-10-15)
Revision as of 11:42, 20 October 2009 by Larsplus (Talk | contribs) (→‎Functional Parameters)

c-Jun promoter (growth factor regulated)

This part is a natural c-Jun promoter and when placed upstream of other genes could induce their transcription in response to Epidermal Growth Factor to the medium.


Usage and Biology

Epidermal Growth Factor (EGF) is a 6045 Da protein discovered by Stanley Cohen in 1986, which won him a Nobel Prize in Physiology and Medicine. EGF regulates cell proliferation by binding to the epidermal growth factor receptors (EGFRs) which are located on the cell surface. Upon binding of EGF to its receptor intrinsic tyrosine kinase activity is stimulated inducing a signaling cascade inside the cell which leads to increased calcium levels, glycolyisis and protein synthesis in the cell. This process ultimately leads to the proliferation of the cell. It was recently shown that c-Jun is one of the targets of EGF action. See [http://2009.igem.org/Team:Heidelberg/Eucaryopedia#EGF Eucaryopedia] for references.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 398
    Illegal BamHI site found at 392
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 110

Functional Parameters

n/ac-Jun promoter (growth factor regulated)

The natural c-Jun promoter was integrated into Part:BBa_K203100 and was induced by Epidermal Growth Factor (100 ng/µl) in HeLa cells. The GFP expression level was determined in [http://2009.igem.org/Team:Heidelberg/Project_Measurement#Two_units_for_promoter_activity_in_mammalian_cells REU] by flow cytometry (1 experiment, 3 replicates) 3 hours after induction.

BBa_K203107 is a natural c-Jun promoter induced more than 100% by Epidermal Growth Factor. The c-Jun promoter/Part:BBa_K203100 construct is coupled to GFP and induced with Epidermal Growth Factor (100 ng/µl) for three hours. The GFP fluorescence of the c-Jun promoter (in [http://2009.igem.org/Team:Heidelberg/Project_Measurement#Two_units_for_promoter_activity_in_mammalian_cells REU]) was measured by flow cytometry once (à three replicates).The standard deviation is indicated by the error bars.


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Parameters
n/ac-Jun promoter (growth factor regulated)