Composite

Part:BBa_K2036017

Designed by: Zhangyu Cheng   Group: iGEM16_HUST-China   (2016-09-20)
Revision as of 17:18, 19 October 2016 by Tianxiongxiao (Talk | contribs)


CI-TT-pR-RBS-GFP-LVAssrAtag

pR is a lytic promoter with two binding sites OR1* and OR2 * which can be recognized by CI. The circuit is built to characterize CI and pR interaction together with control group: pR-GFp-LVAssrAtag (BBa_K2036016).

Fig1:CI and pR interaction characterization circuits

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 1604


Protein&promoter

--CI and pR


CI is a repressor from bacteriophage lambda. To test its interaction with pR promoter, we constructed CI-TT-pR-RBS-GFPLVAssrAtag-PET-Duet-1 and take pR-RBS-GFPLVAssrAtag-PET-Duet-1 as control to test its inhibition function.


Fig2: As the Relative Fluorescent intensity measurement data shows, CI can inhibit pR in minor degree but the leakage expression under pR can’t be ignored, so we should consider to increase the binding sites within pR or the amount of CI coding sequence in the circuit.


Fig3: We also detected GFP reporter in E.coli after induction of 20minute, 120minutes and 240minutes through 20 times of amplification (seen from the figure below). From figure we can find the fluorescence of both two groups was increasing over time and it is obvious that the test group which contains CI expressed less GFP protein than control group. The results verify the inhibition of CI to pR from a more intuitive way.
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