Difference between revisions of "BACTH"
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===Bacterial Adenylate Cyclase-based two hybrid system=== | ===Bacterial Adenylate Cyclase-based two hybrid system=== | ||
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Active adenylate cyclase results in the expression of the ''lacZ'' gene which is detected by blue colonies on plates containing X-Gal. | 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: [http://www.euromedex.com/shop/popup_image.php?pID=208357& | + | see a scheme of the principle: [http://www.euromedex.com/shop/popup_image.php?pID=208357&osCsid=a63d2e53d8afa86ab50e58493b07b400 picture] |
The system consists of four [[plasmid]]s and one E. coli strain (BTH101) | The system consists of four [[plasmid]]s and one E. coli strain (BTH101) | ||
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'''The system:''' | '''The system:''' | ||
| − | + | <pubmed> 9770520 18363795 </pubmed> | |
'''Applications:''' | '''Applications:''' | ||
| − | + | <pubmed> 19400808 19193632 19478798 18363795 20572937 20502438 20933603 </pubmed> | |
Revision as of 04:54, 24 November 2010
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:
Applications:
Frederik M Meyer, Jan Gerwig, Elke Hammer, Christina Herzberg, Fabian M Commichau, Uwe Völker, Jörg Stülke
Physical interactions between tricarboxylic acid cycle enzymes in Bacillus subtilis: evidence for a metabolon.
Metab Eng: 2011, 13(1);18-27
[PubMed:20933603]
[WorldCat.org]
[DOI]
(I p)
Martin Lehnik-Habrink, Henrike Pförtner, Leonie Rempeters, Nico Pietack, Christina Herzberg, Jörg Stülke
The RNA degradosome in Bacillus subtilis: identification of CshA as the major RNA helicase in the multiprotein complex.
Mol Microbiol: 2010, 77(4);958-71
[PubMed:20572937]
[WorldCat.org]
[DOI]
(I p)
Maria A Oliva, Sven Halbedel, Stefan M Freund, Pavel Dutow, Thomas A Leonard, Dmitry B Veprintsev, Leendert W Hamoen, Jan Löwe
Features critical for membrane binding revealed by DivIVA crystal structure.
EMBO J: 2010, 29(12);1988-2001
[PubMed:20502438]
[WorldCat.org]
[DOI]
(I p)
Rok Lenarcic, Sven Halbedel, Loek Visser, Michael Shaw, Ling Juan Wu, Jeff Errington, Davide Marenduzzo, Leendert W Hamoen
Localisation of DivIVA by targeting to negatively curved membranes.
EMBO J: 2009, 28(15);2272-82
[PubMed:19478798]
[WorldCat.org]
[DOI]
(I p)
Denise Lüttmann, Ralf Heermann, Björn Zimmer, Antje Hillmann, Isabella Sofia Rampp, Kirsten Jung, Boris Görke
Stimulation of the potassium sensor KdpD kinase activity by interaction with the phosphotransferase protein IIA(Ntr) in Escherichia coli.
Mol Microbiol: 2009, 72(4);978-94
[PubMed:19400808]
[WorldCat.org]
[DOI]
(I p)
Fabian M Commichau, Fabian M Rothe, Christina Herzberg, Eva Wagner, Daniel Hellwig, Martin Lehnik-Habrink, Elke Hammer, Uwe Völker, Jörg Stülke
Novel activities of glycolytic enzymes in Bacillus subtilis: interactions with essential proteins involved in mRNA processing.
Mol Cell Proteomics: 2009, 8(6);1350-60
[PubMed:19193632]
[WorldCat.org]
[DOI]
(I p)
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)
