Difference between revisions of "Part:BBa K934022"

Revision as of 14:10, 26 September 2012

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”.

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(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]

@@ Line 7: / Line 7: @@
 [[Image:Plux-LasI_result.png|thumb|center|500px|]]
-To characterize Plux-LasI (BBa_K934022), we introduced Plux-LasI (BBa_K934022) with Ptet-LuxR ([https://parts.igem.org/Part:BBa_S03119 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 ([https://parts.igem.org/Part:BBa_K649001 BBa_K649001]) to E.coli as a “Las reporter cell”.
+To characterize Plux-LasI (BBa_K934022), we introduced Plux-LasI (BBa_K934022) with Ptet-LuxR ([https://parts.igem.org/Part:BBa_S03119 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 ([https://parts.igem.org/Part:BBa_K649001 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(K934022) synthesized 3OC12HSL.