Difference between revisions of "Part:BBa K934012:Experience"
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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”. | ||
| − | 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 ( | + | 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. |
We accomplished a positive feedback system with our new Plux-LasI ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K934022 BBa_K934022]). | We accomplished a positive feedback system with our new Plux-LasI ([https://parts.igem.org/wiki/index.php?title=Part:BBa_K934022 BBa_K934022]). | ||
Revision as of 07:39, 26 September 2012
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Applications of BBa_K934012
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”, “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.
We accomplished a positive feedback system with our new Plux-LasI (BBa_K934022).
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.
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