Difference between revisions of "BACTH"
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Revision as of 16:23, 28 May 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:
Applications in B. subtilis:
