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.

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, 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] (P p)


Applications in B. subtilis:

Cori T Leonetti, Matt A Hamada, Stephanie J Laurer, Matthew P Broulidakis, Kyle J Swerdlow, Catherine A Lee, Alan D Grossman, Melanie B Berkmen
Critical Components of the Conjugation Machinery of the Integrative and Conjugative Element ICEBs1 of Bacillus subtilis.
J. Bacteriol.: 2015, 197(15);2558-67
[PubMed:26013486] [WorldCat.org] [DOI] (I p)

Johannes Schneider, Teresa Klein, Benjamin Mielich-Süss, Gudrun Koch, Christian Franke, Oscar P Kuipers, Ákos T Kovács, Markus Sauer, Daniel Lopez
Spatio-temporal remodeling of functional membrane microdomains organizes the signaling networks of a bacterium.
PLoS Genet.: 2015, 11(4);e1005140
[PubMed:25909364] [WorldCat.org] [DOI] (I e)

Lorena Stannek, Martin J Thiele, Till Ischebeck, Katrin Gunka, Elke Hammer, Uwe Völker, Fabian M Commichau
Evidence for synergistic control of glutamate biosynthesis by glutamate dehydrogenases and glutamate in Bacillus subtilis.
Environ. Microbiol.: 2015, 17(9);3379-90
[PubMed:25711804] [WorldCat.org] [DOI] (I p)

Sebastian Dintner, Ralf Heermann, Chong Fang, Kirsten Jung, Susanne Gebhard
A sensory complex consisting of an ATP-binding cassette transporter and a two-component regulatory system controls bacitracin resistance in Bacillus subtilis.
J. Biol. Chem.: 2014, 289(40);27899-910
[PubMed:25118291] [WorldCat.org] [DOI] (I p)

Valerie J Carabetta, Andrew W Tanner, Todd M Greco, Melissa Defrancesco, Ileana M Cristea, David Dubnau
A complex of YlbF, YmcA and YaaT regulates sporulation, competence and biofilm formation by accelerating the phosphorylation of Spo0A.
Mol. Microbiol.: 2013, 88(2);283-300
[PubMed:23490197] [WorldCat.org] [DOI] (I p)

Suey van Baarle, Ilkay Nazli Celik, Karan Gautam Kaval, Marc Bramkamp, Leendert W Hamoen, Sven Halbedel
Protein-protein interaction domains of Bacillus subtilis DivIVA.
J. Bacteriol.: 2013, 195(5);1012-21
[PubMed:23264578] [WorldCat.org] [DOI] (I p)

Haiyan Qiao, Daniela Krajcikova, Chunyan Xing, Baoping Lu, Jinhui Hao, Xi Ke, Hongda Wang, Imrich Barak, Jilin Tang
Study of the interactions between the key spore coat morphogenetic proteins CotE and SpoVID.
J. Struct. Biol.: 2013, 181(2);128-35
[PubMed:23178679] [WorldCat.org] [DOI] (I p)

Martin Lehnik-Habrink, Joseph Newman, Fabian M Rothe, Alexandra S Solovyova, Cecilia Rodrigues, Christina Herzberg, Fabian M Commichau, Richard J Lewis, Jörg Stülke
RNase Y in Bacillus subtilis: a Natively disordered protein that is the functional equivalent of RNase E from Escherichia coli.
J. Bacteriol.: 2011, 193(19);5431-41
[PubMed:21803996] [WorldCat.org] [DOI] (I p)

Christelle M Roux, Jonathon P DeMuth, Paul M Dunman
Characterization of components of the Staphylococcus aureus mRNA degradosome holoenzyme-like complex.
J. Bacteriol.: 2011, 193(19);5520-6
[PubMed:21764917] [WorldCat.org] [DOI] (I p)

Kenneth Briley, Peter Prepiak, Miguel J Dias, Jeanette Hahn, David Dubnau
Maf acts downstream of ComGA to arrest cell division in competent cells of B. subtilis.
Mol. Microbiol.: 2011, 81(1);23-39
[PubMed:21564336] [WorldCat.org] [DOI] (I p)

Tatsuya Fukushima, Isako Furihata, Robyn Emmins, Richard A Daniel, James A Hoch, Hendrik Szurmant
A role for the essential YycG sensor histidine kinase in sensing cell division.
Mol. Microbiol.: 2011, 79(2);503-22
[PubMed:21219466] [WorldCat.org] [DOI] (I p)

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)

Inga Wadenpohl, Marc Bramkamp
DivIC stabilizes FtsL against RasP cleavage.
J. Bacteriol.: 2010, 192(19);5260-3
[PubMed:20644139] [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)

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)

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