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&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 [[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 23:48, 23 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: <pubmed> 9770520 18363795 </pubmed>
Applications: <pubmed> 19400808 19193632 19478798 18363795 20572937 20502438 20933603 </pubmed>
