Regulatory

Part:BBa_K203114:Design

Designed by: Lars Velten, Simon Haas, Anne Rademacher and Hannah Meyer   Group: iGEM09_Heidelberg   (2009-10-15)
Revision as of 10:14, 17 October 2009 by Larsplus (Talk | contribs) (→‎References)

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PPARγ-regulated promoter 1


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]


Design Notes

[http://2009.igem.org/Team:Heidelberg/Project_Synthetic_promoters RA-PCR] (Fig. 1) was conducted with Oligos containing a pPARγ binding site, plus a small number of "general activators" (NF-Y, Sp1, Ap1, CREB). 12 clones were picked, miniprepped and transfected. pPARγ was then induced by the addition of Thiazolidinedione (5µM) for 10 hours, and left uninduced as a control. The plate was then scanned by TECAN, an automated fluorescence plate reader. TECAN is very imprecise on eukaryotic cells, and the arbitrary fluorescence we meausred is not proportional to [http://2009.igem.org/Team:Heidelberg/Project_Measurement REU] or another precise measure of mammalian promoter activity, but it can serve as a rough indicator of promoter induction. We picked two promising clones for submission to the registry and provide a rough characterization.

Fig. 1: The method of LAM-PCR. For a detailled description, see [http://2009.igem.org/Team:Heidelberg/Project_Synthetic_promoters our wiki]


Source

Synthesized in our laboratory.

References

RA-PCR, a method for the generation of randomized promoter libraries. igem 2009 Heidelberg team wiki. Available online at http://2009.igem.org/Team:Heidelberg/Project_Synthetic_promoters#Results