Regulatory

Part:BBa_K203114

Designed by: Lars Velten, Simon Haas, Anne Rademacher and Hannah Meyer   Group: iGEM09_Heidelberg   (2009-10-15)
Revision as of 10:08, 17 October 2009 by Larsplus (Talk | contribs) (→‎Functional Parameters)

PPARγ-regulated promoter 1

A synthetic promoter upregulated by pPARγ activation. Manufactured by [http://2009.igem.org/Team:Heidelberg/Project_Synthetic_promoters RA-PCR].

Usage and Biology

pPARγ activation. During fatty diet, fatty acids are converted to prostaglandins by oxygenases, which activate pPARγ. Image published under [http://en.wikipedia.org/wiki/File:PPAR-diagram.png GNU Free documentation license.]
Peroxisome proliferator-activated receptor γ (PPAR γ) is a transcription factor belonging to the family of nuclear receptors. PPAR γ plays an important role in glucose metabolism and fatty acid storage. PPAR γ is basically activated by ligands like the prostaglandin PGJ2 and through dimerization with retinoid X receptor (RXR). The activated heterodimer binds to the DNA consensus sequence AGGTCANAGGTCA resulting in an increased or decreased transcription of the appropriate gene. The genes activated by PPAR γ initiate the uptake of fatty acids and differentiation of cells to adipocytes. Besides its function in metabolism, PPAR γ was also shown to be correlated with several diseases such as cancer and diabetes. Activation of PPAR γ by synthetic PPAR γ ligands result in an increased glucose uptake. These syntethtic ligands are therefore promising agents in diabetes II treatment. Another synthetic ligand of PPAR γ is able to inhibit the proliferation of different cancer types. For references, see [http://2009.igem.org/Team:Heidelberg/Eucaryopedia Heidelberg 2009's Eukaryopedia].


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Functional Parameters

pPARγ was induced by 5µM Thiazolidinedione in [http://2009.igem.org/Team:Heidelberg/Eucaryopedia#U2-OS U2OS cells]. Promoter activtiy was then roughly characterized in [http://2009.igem.org/Team:Heidelberg/Measurement REU] by TECAN (automated plate fluorescence reader). This promoter corresponds to clone γL9.

Characterization of two pPARγ responsive promoters. Clones created by [http://2009.igem.org/Team:Heidelberg/Project_Synthetic_promoters RA-PCR] were screened and two promising clones were characterized by triple TECAN reads.


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Categories
Parameters
None