Difference between revisions of "Part:BBa K243028"

Latest revision as of 23:42, 21 October 2009

FOS-Split Linker-Fok_a

Activator protein-1 (AP-1) is a crucial transcription factor implicated in numerous cancers and binds DNA as a dimer. Two classes of core DNA sequences, the sequences TRE (TGACTCA) and CRE (TGACGTCA), can be recognized by the AP-1. Nine homologues of the AP-1 leucine zipper region have been characterized. All of them are able to form heterodimers, some also form homodimers. One of them is the so called c-Fos. Via leucin zipper they interact among each other and with their bipartite domain they bind DNA. (Abate et al., Mol Cell Biol. 1991 July).

Heterodimeric Fos complex (http://www.rcsb.org/pdb/explore/images.do?structureId=1FOS)

Usage and Biology

To avoid the labeling of oligos we tried an alternative way of binding of the heterodimeric Fok to the DNA by using the DNA binding domain of Fos as an adapter between the Fok construct and the DNA. For our needs, we chose the natural bZIP sequence of Fos and Jun.

We fused Split Linker-Fok_a to Fos.The mixture of this construct with another hybrid protein called Jun allows the dimerization of Fos and therefore its binding at their target sequence TRE (TGACTCA) or CRE (TGACGTCA) by using the bZIP sequence of Jun as adapter. The addition of Fok_i leads to DNA cleavage.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 19
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 766

@@ Line 12: / Line 12: @@
 ===Usage and Biology===
-To avoid the labeling of oligos we tried an alternative way of binding of the heterodimeric Fok to the DNA, by using the DNA binding domain of Fos as adapter between the Fok construct and the DNA.  For our needs, we chose the natural bZIP sequence of Fos and Jun.
+To avoid the labeling of oligos we tried an alternative way of binding of the heterodimeric Fok to the DNA by using the DNA binding domain of Fos as an adapter between the Fok construct and the DNA. For our needs, we chose the natural bZIP sequence of Fos and Jun.
-Split Linker-Fok_a can be fused to Fos. The mixture of this construct with another hybrid protein Jun allows the dimerization of Fos and therefore its binding at their target sequence TRE (TGACTCA) or CRE (TGACGTCA) by using the bZIP sequence of Jun as adapter. The addition of Fok_i leads to DNA cleavage.
+We fused Split Linker-Fok_a to Fos.The mixture of this construct with another hybrid protein called Jun allows the dimerization of Fos and therefore its binding at their target sequence TRE (TGACTCA) or CRE (TGACGTCA) by using the bZIP sequence of Jun as adapter. The addition of Fok_i leads to DNA  cleavage.
-[[Image:Freiburg 09 JunFos new2.jpg|250x330px]]<br>
+[[Image:Freiburg 09 fos jun.jpg|250x330px]]<br>