Part:BBa_K934022

Plux-LasI

We constructed this part by combining BBa_R0062 and BBa_K081016. This part produces LasI enzyme in the presence of LuxR-3OC6HSL complex.

To characterize Plux-LasI (BBa_K934022), we introduced Plux-LasI (BBa_K934022) with Ptet-LuxR (BBa_S03119) to E.coli as “3OC6HSL dependent 3OC12HSL producer cell”. In this E.coli, constitutively expressed LuxR activates the expression of LasI in the presence of 3OC6HSL. We then introduced Ptrc-LasR and Plas-GFP (BBa_K649001) to E.coli as a “Las reporter cell”.

When the supernatants of the culture of “3OC6HSL dependent 3OC12HSL producer cell” were induced to “Las reporter cell”, GFP expression in “Las reporter cell” was activated. Thus, the expression of GFP in “Las reporter cell” is dually regulated by 3OC12HSL produced by “3OC6HSL dependent 3OC12HSL producer cell”, this result shows that Plux-lasI(K934022) synthesized 3OC12HSL.

We accomplished a positive feedback system with our improved part Plux-LasI(BBa_K934012).

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 745
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 307
  
• 1000
  COMPATIBLE WITH RFC[1000]