Difference between revisions of "Part:BBa K3720003"
 
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<partinfo>BBa_K3720003 short</partinfo>
 
<partinfo>BBa_K3720003 short</partinfo>
  
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When used in combination with CooA, it is possible to create a light emitting mechanism that reacts with carbon monoxide.
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pCooM is CooA-dependent promoter originates from <i>Rhodospirillum rubrum</i>. Since CooA binds to carbon monoxide and activates pCooM, you can create a light-emitting mechanism by using this part with CooA.
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Culture pBEST incorporating this part in LD medium and transfer 20㎕ to a 1.5-ml tube.
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Then, 0.5㎕ of D-luciferin was added, CO was injected, and luminescence was measured.
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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.
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Quantitative measurements were not possible because the laboratory facilities did not allow us to prepare a luminometer.
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As shown in the photo, the pipette which carbon monoxide was injected showed a change in color.
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<p>[[File:T--Qdai--result.png|500px|thumb|center|Fig.1 Color changes in bacterial culture <br>Left: LB medium+luciferin&nbsp;&nbsp;&nbsp;&nbsp;Right: LB medium+luciferin+CO gas]] </p>
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<p>[[File:T--Qdai--result.png|500px|thumb|center|figure.1 Color changes in bacterial culture <br>Left: LB medium+luciferin Right: LB medium+luciferin+CO gas]] </p>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 03:03, 22 October 2021


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