Difference between revisions of "Part:BBa K1946004"

Revision as of 19:37, 25 October 2016

Toluene/Xylene Sensor

This part responds to the presence of toluene, m-, p- or o-xylenes and produces a green fluorescent signal. The transcriptional regulatory protein XylR recognizes these organic molecules and activates pu promoter which in turn will lead to the synthesis of sfGFP.

We cloned our XylR after pL(TetO) promoter which is operated by TetR and sfGFP after pu promoter into pet22b vector. We confirmed our cloning with restriction digestion with SpeI and XbaI.

We expect that in the presence of ATC, XylR will be expressed and after XylR expression, in the presence of xylenes or toluene sfGFP will be expressed. We induced our cells ATC and, after 2 hours of incubation, with xylene at different concentrations. Unfortunately, we have not seen any difference in sfGFP signal.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 2094
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 2248
Illegal BamHI site found at 1751
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 1023
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1168
Illegal BsaI.rc site found at 1546
Illegal SapI.rc site found at 2266

@@ Line 6: / Line 6: @@
+We cloned our XylR after pL(TetO) promoter which is operated by TetR and sfGFP after pu promoter into pet22b vector. We confirmed our cloning with restriction digestion with SpeI and XbaI.
+[[File:BBa_K1946004_xylr_digestion.jpeg|500px|thumb|center]]
+We expect that in the presence of ATC, XylR will be expressed and after XylR expression, in the presence of xylenes or toluene sfGFP will be expressed. We induced our cells ATC and, after 2 hours of incubation, with xylene at different concentrations. Unfortunately, we have not seen any difference in sfGFP signal.
 [[File:T--Bilkent-UNAMBG--xylR.png|500px|thumb|center]]
-We cloned our XylR after pL(TetO) promoter which is operated by TetR and sfGFP after pu promoter into pet22b vector. We expect that in the presence of ATC, XylR will be expressed and after XylR expression, in the presence of xylenes or toluene sfGFP will be expressed. We induced our cells ATC and, after 2 hours of incubation, with xylene at different concentrations. Unfortunately, we have not seen any difference in sfGFP signal.