Difference between revisions of "BACTH"
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| − | + | ===Bacterial Adenylate Cyclase-based two hybrid system=== | |
Allows the detection of in vivo protein-protein interactions by restoring the activity of the ''Bordetella pertussis'' adenylate cyclase in ''E. coli'' reporter strains. | Allows the detection of in vivo protein-protein interactions by restoring the activity of the ''Bordetella pertussis'' adenylate cyclase in ''E. coli'' reporter strains. | ||
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see a scheme of the principle: [http://www.euromedex.com/shop/popup_image.php?pID=208357&osCsid=a63d2e53d8afa86ab50e58493b07b400 picture] | see a scheme of the principle: [http://www.euromedex.com/shop/popup_image.php?pID=208357&osCsid=a63d2e53d8afa86ab50e58493b07b400 picture] | ||
| − | The system consists of four | + | The system consists of four [[plasmid]]s and one E. coli strain (BTH101) |
| − | # pKT25: C-terminal fusion of the target protein | + | # pKT25: C-terminal fusion of the target protein to the T25 fragment of adenylate cyclase |
| − | # pKT25-N: N-terminal fusion of the target protein | + | # pKT25-N: N-terminal fusion of the target protein to the T25 fragment of adenylate cyclase |
| − | # pUT18: N-terminal fusion of the target protein | + | # pUT18: N-terminal fusion of the target protein to the T18 fragment of adenylate cyclase |
| − | # pUT18C: C-terminal fusion of the target protein | + | # pUT18C: C-terminal fusion of the target protein to the T18 fragment of adenylate cyclase |
| + | |||
| + | E. coli BTH101 is a cya mutant that requires [[plasmid]]-derived adenylate cyclase activity for the activation of the ''lac'' operon. | ||
| + | |||
| + | The BACTH is commercially available from [http://www.euromedex.com/shop/pdf/produits/BACTH-Manual.pdf?osCsid=a63d2e53d8afa86ab50e58493b07b400 EUROMEDEX] | ||
| + | |||
| + | ===References:=== | ||
| + | |||
| + | '''The system:''' | ||
| + | <pubmed>9576956 18363795 </pubmed> | ||
| + | |||
| + | '''Applications in ''B. subtilis'':''' | ||
| + | <pubmed> 19193632 19478798 18363795 20572937 20502438 20933603 20644139, 21219466 21564336 21764917 21803996 23178679 23264578 23490197 26013486 25118291,25909364 25711804</pubmed> | ||
Latest revision as of 17:14, 2 December 2015
Bacterial Adenylate Cyclase-based two hybrid system
Allows the detection of in vivo protein-protein interactions by restoring the activity of the Bordetella pertussis adenylate cyclase in E. coli reporter strains.
Active adenylate cyclase results in the expression of the lacZ gene which is detected by blue colonies on plates containing X-Gal.
see a scheme of the principle: picture
The system consists of four plasmids and one E. coli strain (BTH101)
- pKT25: C-terminal fusion of the target protein to the T25 fragment of adenylate cyclase
- pKT25-N: N-terminal fusion of the target protein to the T25 fragment of adenylate cyclase
- pUT18: N-terminal fusion of the target protein to the T18 fragment of adenylate cyclase
- pUT18C: C-terminal fusion of the target protein to the T18 fragment of adenylate cyclase
E. coli BTH101 is a cya mutant that requires plasmid-derived adenylate cyclase activity for the activation of the lac operon.
The BACTH is commercially available from EUROMEDEX
References:
The system:
Dennis Claessen, Robyn Emmins, Leendert W Hamoen, Richard A Daniel, Jeff Errington, David H Edwards
Control of the cell elongation-division cycle by shuttling of PBP1 protein in Bacillus subtilis.
Mol Microbiol: 2008, 68(4);1029-46
[PubMed:18363795]
[WorldCat.org]
[DOI]
(I p)
G Karimova, J Pidoux, A Ullmann, D Ladant
A bacterial two-hybrid system based on a reconstituted signal transduction pathway.
Proc Natl Acad Sci U S A: 1998, 95(10);5752-6
[PubMed:9576956]
[WorldCat.org]
[DOI]
(P p)
Applications in B. subtilis:
