Difference between revisions of "Part:BBa K934012"
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[[Image:Plas-LuxI_result.png|thumb|center|500px|This work was done by M.M, X.T and S.L.]]
 
[[Image:Plas-LuxI_result.png|thumb|center|500px|This work was done by M.M, X.T and S.L.]]
  
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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”.
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To characterize Plas-LuxI (BBa_K934012), we introduced Plas-LuxI on pSB3K3 with Ptrc-LasR on pSB6A1 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”.
  
 
When the supernatants of the culture of “3OC12HSL dependent 3OC6HSL producer cell” were induced to “Lux reporter cell”, GFP expression in “Lux reporter cell” was activated. This result shows that Plas-LuxI (BBa_K934012) is fuctioning.
 
When the supernatants of the culture of “3OC12HSL dependent 3OC6HSL producer cell” were induced to “Lux reporter cell”, GFP expression in “Lux reporter cell” was activated. This result shows that Plas-LuxI (BBa_K934012) is fuctioning.

Revision as of 08:55, 5 October 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.


This work was done by M.M, X.T and S.L.

To characterize Plas-LuxI (BBa_K934012), we introduced Plas-LuxI on pSB3K3 with Ptrc-LasR on pSB6A1 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”.

When the supernatants of the culture of “3OC12HSL dependent 3OC6HSL producer cell” were induced to “Lux reporter cell”, GFP expression in “Lux reporter cell” was activated. This result shows that Plas-LuxI (BBa_K934012) is fuctioning.

We improved a previous part Plas-LuxI (BBa_K266000)

and accomplished a positive feedback system with our new part Plux-LasI (BBa_K934022).


This work was done by M.M, X.T and S.L.

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]