Composite

Part:BBa_K3594003

Designed by: Runhan Luo   Group: iGEM20_SHSBNU_China   (2020-10-23)
Revision as of 19:15, 23 October 2020 by GraceLuo (Talk | contribs)

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Quorum Sensing Fluorescence Reporting Device

This composite part is used as the reporter of the quorum sensing device. It consists a Plux promotor and BBa_K3594009, a sfGFP gene. When the concentration of the signal molecule-LuxR-AHL reaches threshold, the bacteria start to commit suicide and at the same time, emit large amount of fluorescence.

synchronized lysis circuit in M.Omar Din et al. Nature, 2016

In our project, we made the following changes to the above experiment:

the design of our bacterial synchronized lysis circuit

pSHS6_pSB1C3_pTac_LuxR_plux_sfGFP: constructed by Gibson assembly T4 Ligation: pTac_LuxR_plux_sfGFP(~3.6Kb)and pSB1C3 backbone(~2Kb) This plasmid is used to test if LuxR and plux promoter work well.

the electrophoretogram of colony PCR of BBa K3594003

Verification of LuxR and Plux

Sequencing results of plux-GFP
The result of BBa_K3594003

If the fluorescence value in the tube increases with time, and the fluorescence value in the tube with IPTG and atc is higher than no inducer or only one inducer, it proves that the bacteria emit fluorescence after reaching the threshold, therefore the experiment is successful. Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1208
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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