Generator

Part:BBa_K511819

Designed by: Grant Robinson   Group: iGEM11_MIT   (2011-09-28)
Revision as of 16:00, 5 November 2011 by Chsuqed (Talk | contribs)

Inducible Yellow Fluorescent Protein Generator (UAS-Gal4-Citrine) MammoBlock Device

This MammoBlock composite device produces the yellow/green fluorescent protein Citrine when induced with Gal4 transactivator variants and otherwise produces Citrine at a low, basal (OFF) level.


Another activator promoter pair we make use of is the Gal4-UAS system. Gal4VP16 is an activator capable of binding to UAS promoter site and activating expression of downstream genes. Here we characterize this interaction with a set of transfections. Cells were transfected with UAS:(mKate, eYFP-FF4, eBFP2, or H2B-citrine) and Hef1a:GV16. Controls lack the Hef1a:GV16 plasmid. The above graph displays mean fluoresence in the denoted channels.

As can be clearly seen, Hef1a:GV16 results in significant activation of the UAS promoter. This results in the observed increases in mean fluoresence compared to control populations. With the exception of UAS:eBFP2, we see more than 20-fold increase in mean fluorescence.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 126
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 126
    Illegal NheI site found at 154
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 126
    Illegal XhoI site found at 83
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 126
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 126
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
n/aInducible Yellow Fluorescent Protein Generator (UAS-Gal4-Citrine) MammoBlock Device