Difference between revisions of "Part:BBa K934012"
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To characterize Plas-LuxI (BBa_K934012), we introduced Plas-LuxI (BBa_K934012) with Ptrc-LasR to E.coli as “3OC12HSL dependent 3OC6HSL producer cell”. In this E.coli, constitutively expressed LasR activates the expression of LuxI in the presence of 3OC12HSL. We then introduced Ptet-LuxR ([https://parts.igem.org/Part:BBa_S03119 BBa_S03119]) and Plux-GFP ([https://parts.igem.org/Part:BBa_K395100 BBa_K395100]) to E.coli as a “Lux reporter cell”.
 
To characterize Plas-LuxI (BBa_K934012), we introduced Plas-LuxI (BBa_K934012) with Ptrc-LasR to E.coli as “3OC12HSL dependent 3OC6HSL producer cell”. In this E.coli, constitutively expressed LasR activates the expression of LuxI in the presence of 3OC12HSL. We then introduced Ptet-LuxR ([https://parts.igem.org/Part:BBa_S03119 BBa_S03119]) and Plux-GFP ([https://parts.igem.org/Part:BBa_K395100 BBa_K395100]) to E.coli as a “Lux reporter cell”.
  
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In the presence of 3OC6HSL produced by “3OC12HSL dependent 3OC6HSL producer cell”, “Lux reporter cell” was activated and GFP was expressed. Thus, the expression of GFP in “Lux reporter cell” is dually regulated by 3OC6HSL produced by “3OC12HSL dependent 3OC6HSL producer cell”, this result shows that Plas-LuxI (BBa_K934012) synthesized 3OC6HSL.
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In the presence of 3OC6HSL produced by “3OC12HSL dependent 3OC6HSL producer cell”, GFP expression in “Lux reporter cell” was activated. Thus, the expression of GFP in “Lux reporter cell” is dually regulated by 3OC6HSL produced by “3OC12HSL dependent 3OC6HSL producer cell”, this result shows that Plas-LuxI (BBa_K934012) synthesized 3OC6HSL.
  
 
We improved a previous part Plas-LuxI ([https://parts.igem.org/Part:BBa_K266000:Experience#User_Reviews BBa_K266000])  
 
We improved a previous part Plas-LuxI ([https://parts.igem.org/Part:BBa_K266000:Experience#User_Reviews BBa_K266000])  

Revision as of 12:45, 26 September 2012

Plas-LuxI

We constructed this part by combining BBa_K649000 and BBa_K081008. This part generates LuxI enzyme in the presence of LasR-3OC12HSL complex.


Plas-LuxI result.png

To characterize Plas-LuxI (BBa_K934012), we introduced Plas-LuxI (BBa_K934012) with Ptrc-LasR to E.coli as “3OC12HSL dependent 3OC6HSL producer cell”. In this E.coli, constitutively expressed LasR activates the expression of LuxI in the presence of 3OC12HSL. We then introduced Ptet-LuxR (BBa_S03119) and Plux-GFP (BBa_K395100) to E.coli as a “Lux reporter cell”.

In the presence of 3OC6HSL produced by “3OC12HSL dependent 3OC6HSL producer cell”, GFP expression in “Lux reporter cell” was activated. Thus, the expression of GFP in “Lux reporter cell” is dually regulated by 3OC6HSL produced by “3OC12HSL dependent 3OC6HSL producer cell”, this result shows that Plas-LuxI (BBa_K934012) synthesized 3OC6HSL.

We improved a previous part Plas-LuxI (BBa_K266000) and accomplished a positive feedback system with our new Plux-LasI (BBa_K934022).


For more information, see [http://2012.igem.org/Team:Tokyo_Tech/Project our work in Tokyo_Tech 2012 wiki].

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 749
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]