Device

Part:BBa_K1946004

Designed by: Ayşe Naz Ozantürk   Group: iGEM16_Bilkent-UNAMBG   (2016-10-19)


Toluene/Xylene Sensor

This part responds to the presence of toluene, m-, p- or o-xylenes and produces a green fluorescent signal. The transcriptional regulatory protein XylR recognizes these organic molecules and activates pu promoter which in turn will lead to the synthesis of sfGFP.


We cloned our XylR after pL(TetO) promoter which is operated by TetR and sfGFP after pu promoter into pet22b vector. We confirmed our cloning with restriction digestion with SpeI and XbaI.

BBa K1946004 xylr digestion.jpeg

We expect that in the presence of ATC, XylR will be expressed and after XylR expression, in the presence of xylenes or toluene sfGFP will be expressed. We induced our cells ATC and, after 2 hours of incubation, with xylene at different concentrations. Unfortunately, we have not seen any difference in sfGFP signal.

T--Bilkent-UNAMBG--xylR.png


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2094
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2248
    Illegal BamHI site found at 1751
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1023
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1168
    Illegal BsaI.rc site found at 1546
    Illegal SapI.rc site found at 2266


[edit]
Categories
//function/sensor
Parameters
device_typexylene sensor