Part:BBa_K934022:Experience
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Applications of BBa_K934022
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 (BBa_K934011) and Plas-GFP (BBa_K649001) to E.coli as a “Las reporter cell”.
In the presence of 3OC12HSL produced by “3OC6HSL dependent 3OC12HSL producer cell”, “Las reporter cell” was activated and GFP was expressed. 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 (BBa_K934022) synthesized 3OC12HSL.
We accomplished a positive feedback system with our improved part Plux-LasI (BBa_K934012).
By co-culturing Plux-LasI cell and Plas-LuxI cell, we confirmed higher concentration of a signal than initial conditions was detected through production of the other signal (red arrow & blue arrow). This result strongly suggests that our positive feedback system worked accurately.
User Reviews
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