Difference between revisions of "Part:BBa K934022"

Revision as of 08:58, 5 October 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.

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

To characterize Plux-LasI (BBa_K934022), we introduced Plux-LasI pSB3K3 with Ptet-LuxR (BBa_S03119) on pSB6A1 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. This result shows that Plux-lasI(K934022) is functioning.

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

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
INCOMPATIBLE WITH RFC[10]
Unknown
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|This work was done by M.M, X.T and S.L.]]
-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 pSB3K3 with Ptet-LuxR ([https://parts.igem.org/Part:BBa_S03119 BBa_S03119]) on pSB6A1 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”.
 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. This result shows that Plux-lasI(K934022) is functioning.