Composite

Part:BBa_K3720003

Designed by: Kazuma Tsurumaki   Group: iGEM21_Qdai   (2021-10-01)


Firefly luciferase with pCooM, RBS (weak), and double terminator

pCooM is CooA-dependent promoter originates from Rhodospirillum rubrum. Since CooA binds to carbon monoxide and activates pCooM, you can create a light-emitting mechanism by using this part with CooA.

We introduced a plasmid incorporating this part into our E. coli K-12 strain. Then, we mixed 40㎕ of liquid culture medium of this E. coli with 1.0㎕ of D-luciferin (15 mg/ml) and subsequently injected CO and measured the luminescence. Although quantitative measurements could not be conducted because there is no proper device in our lab for observing bioluminescence. However, as shown in the picture, we can see with the naked eye that the color of the tube injected with carbon monoxide has changed.

Fig.1 Color changes in bacterial culture
Left: LB medium+luciferin    Right: LB medium+luciferin+CO gas

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 SapI.rc site found at 900


[edit]
Categories
//chassis/prokaryote/ecoli
//classic/reporter
//function/reporter/light
Parameters
chassisE. coli K12
colorGreen
controlCooA
device_typereporter
directionForward
outputluciferase