Device

Part:BBa_K317039

Designed by: Yasuhiko SASAKI   Group: iGEM10_Tokyo-NoKoGen   (2010-10-21)

Pconst.(H)-RBS-LuxR-Double term-Plux-Lysis

This device senses the AHL concentration in the media and regulates the expression of downstream genes. The complex of the LuxR protein and the AHL activates the promoter, increasing the rate of transcription. Also, This devicce includes BBa_K112808 at the downstream gene, the auto lysis device which causing lysis of E.coli


Evaluation of BBa_K317039

We performed a experiment with this device. The experiment was performed from single colonies in a streaked LB agar plate.At first, we cultured the single colony having the above device in 100 mL LB for 6 hors at 37℃ with agitation.Then, the culture was dispensed into 3 mL scale and AHL(f.c. 50 nM) was added to each dispensed culture to induce the expression of Lysis genes. The obtained culture was incubated for 20 hours at room temperature without agitation. After that, the OD660 of each culture(N=3) was measured.The result was shown below.


AHL Lysis.png



The result showed that the OD660 were different in presence of AHL(red bar) or not (gray bar). OD660 of samples with the AHL were half less than no additive samples. That indicates that the complex of AHL and LuxR activated the expression of lysis genes and the E.coli in the culture were lysed.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 2400
    Illegal NheI site found at 2423
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 2005
    Illegal AgeI site found at 2075
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 985
    Illegal SapI site found at 2656


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