Difference between revisions of "Part:BBa K511823"

 
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This MammoBlock composite device produces the monomeric red fluorescent protein mKate when induced with Gal4 transactivator variants and otherwise produces mKate at a low, basal (OFF) level.
 
This MammoBlock composite device produces the monomeric red fluorescent protein mKate when induced with Gal4 transactivator variants and otherwise produces mKate at a low, basal (OFF) level.
  
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<html><img src='https://static.igem.org/mediawiki/2011/thumb/6/66/Hef1a-GV16%2C_UAS-Reporter_Data.jpg/800px-Hef1a-GV16%2C_UAS-Reporter_Data.jpg
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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.
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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.
  
 
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Revision as of 04:25, 29 September 2011

Inducible Red Fluorescent Protein Generator (UAS-Gal4-mKate) MammoBlock Device

This MammoBlock composite device produces the monomeric red fluorescent protein mKate when induced with Gal4 transactivator variants and otherwise produces mKate 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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 876
    Illegal SapI.rc site found at 258