Composite

Part:BBa_K2036010

Designed by: Zhangyu Cheng   Group: iGEM16_HUST-China   (2016-09-19)
Revision as of 16:32, 19 October 2016 by Tianxiongxiao (Talk | contribs)


Cro-TT-pRM-RBS-GFP-LVAssrAtag

It is a negtive control of GFP expression. When inserted into a expression plasmid behind an inducible promoter,it will form a NO gate. Take PETDuet-1 as an example,after constructing PETDurt-1-Cro-TT-pRM-RBS-GFP-LVAssrAtag,IPTG can trigger T7 promoter in the plasmid, and cro can bind to a certain site within pRM, so that GFP will not be produced. And we get a NO gate of GFP generator.
HUST-Chian 2016 Build this circuit to test Cro and pRM interaction intensity with contol group:pRM-GFP-LVAssrAtag (BBa_K2036009)

Fig1:Cro&pRM interaction characterization circuit

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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1036


Protein&promoter

--Cro and pRM


Fig2: We characterized cro and pRM inhibition by the same method as CI and pR’s. From line chart and fluorescence detection, we can see that the test group contains cro expressed less GFP protein than control group over time. It proves that cro can effectively bind pRM to block its downstream gene’s transcription.


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Categories
Parameters
None