Difference between revisions of "Part:BBa K909007"

Revision as of 00:24, 27 September 2012

TetR DNA binding domain containing BamHI site for protein fusions

Truncated version of tetracycline repressor TetR, consisting of 1 – 127 amino acids, containing DNA binding domain. The rest of the protein, responsible for tetR dimerization, were removed. Without dimerization domain, tetR-DBD fails to bind TetR responsive promoter efficiently and unable to repress transcription of reporter protein (see figure). However, TetR-DBD fusion with dimerazing proteins, e.g. UVR8 protein (BBa_K909008), restores TetR-DBD promoter binding.

GFP repression (reporter) by full length TetR, TetR-DBD and dimerizing TetR-DBD-UVR8 fusion

Usage and Biology

We introduced BamHI site at C terminus of tetR-DBD for an easy in frame fusions with other proteins. This can easily be used as a two hybrid system to detect homo/heterodimers in E. coli, and, in principle, one can use these fusions to turn protein-protein interaction into the repression of transcription. Furthermore, inducible dimerization can lead to a novel switch like behaviors of the system (e.g. [http://2012.igem.org/Team:ETH_Zurich/UVR8 ETH Zurich 2012 team] created a novel UV-B inducible-ON switch).

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 382
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
-Truncated version of tetracycline repressor TetR, consisting of 1 – 127 amino acids, containing DNA binding domain. The rest of the protein, alpha 8-10 helices responsible for tetR dimerization. Without dimerization domain, tetR-DBD fails to bind TetR responsive promoter efficiently and unable to repress transcription of reporter protein (see figure).  However, TetR-DBD fusion with UVR8 protein (<partinfo>BBa_K909008</partinfo>) restores TetR-DBD promoter binding.
+Truncated version of tetracycline repressor TetR, consisting of 1 – 127 amino acids, containing DNA binding domain. The rest of the protein, responsible for tetR dimerization, were removed. Without dimerization domain, tetR-DBD fails to bind TetR responsive promoter efficiently and unable to repress transcription of reporter protein (see figure). However, TetR-DBD fusion with dimerazing proteins, e.g. UVR8 protein (<partinfo>BBa_K909008</partinfo>), restores TetR-DBD promoter binding.
 [[Image:TetR, tetR-DBD, tetR-DBD-UVR8 represion.jpg|frameless|upright=2.5|center|thumb|GFP repression (reporter) by full length TetR, TetR-DBD and dimerizing TetR-DBD-UVR8 fusion
@@ Line 11: / Line 11: @@
 ===Usage and Biology===
-We introduced BamHI site at C terminus of tetR-DBD for an easy in frame fusions with other proteins. This can easily be used as a two hybrid system to detect homo/heterodimers in E. coli, and, in principle, one can use these fusions to turn protein-protein interaction into the repression of transcription. Furthermore, inducible dimerization can lead to a novel switch like behaviors of the system (e.g. [http://2012.igem.org/Team:ETH_Zurich ETH Zurich 2012 team] created a novel UV-B inducible-ON switch).
+We introduced BamHI site at C terminus of tetR-DBD for an easy in frame fusions with other proteins. This can easily be used as a two hybrid system to detect homo/heterodimers in ''E. coli'', and, in principle, one can use these fusions to turn protein-protein interaction into the repression of transcription. Furthermore, inducible dimerization can lead to a novel switch like behaviors of the system (e.g. [http://2012.igem.org/Team:ETH_Zurich/UVR8 ETH Zurich 2012 team] created a novel UV-B inducible-ON switch).