BACTH

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)

1. pKT25: C-terminal fusion of the target protein to the T25 fragment of adenylate cyclase
2. pKT25-N: N-terminal fusion of the target protein to the T25 fragment of adenylate cyclase
3. pUT18: N-terminal fusion of the target protein to the T18 fragment of adenylate cyclase
4. 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, C Fayolle, S Gmira, A Ullmann, C Leclerc, D Ladant
Charge-dependent translocation of Bordetella pertussis adenylate cyclase toxin into eukaryotic cells: implication for the in vivo delivery of CD8(+) T cell epitopes into antigen-presenting cells.
Proc Natl Acad Sci U S A: 1998, 95(21);12532-7
[PubMed:9770520] [WorldCat.org] [DOI] (P p)

Applications:

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)

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)