Chapter title |
Protein–Protein Interaction: Bacterial Two-Hybrid
|
---|---|
Chapter number | 13 |
Book title |
Bacterial Protein Secretion Systems
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7033-9_13 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7031-5, 978-1-4939-7033-9
|
Authors |
Gouzel Karimova, Emilie Gauliard, Marilyne Davi, Scot P. Ouellette, Daniel Ladant |
Editors |
Laure Journet, Eric Cascales |
Abstract |
The bacterial two-hybrid (BACTH, for "Bacterial Adenylate Cyclase-Based Two-Hybrid") system is a simple and fast genetic approach to detecting and characterizing protein-protein interactions in vivo. This system is based on the interaction-mediated reconstitution of a cyclic adenosine monophosphate (cAMP) signaling cascade in Escherichia coli. As BACTH uses a diffusible cAMP messenger molecule, the physical association between the two interacting chimeric proteins can be spatially separated from the transcription activation readout, and therefore it is possible to analyze protein-protein interactions that occur either in the cytosol or at the inner membrane level as well as those that involve DNA-binding proteins. Moreover, proteins of bacterial origin can be studied in an environment similar (or identical) to their native one. The BACTH system may thus permit a simultaneous functional analysis of proteins of interest-provided the hybrid proteins retain their activity and their association state. This chapter describes the principle of the BACTH genetic system and the general procedures to study protein-protein interactions in vivo in E. coli. |
Mendeley readers
Geographical breakdown
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Unknown | 81 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 17 | 21% |
Student > Bachelor | 8 | 10% |
Student > Master | 8 | 10% |
Researcher | 7 | 9% |
Student > Doctoral Student | 4 | 5% |
Other | 8 | 10% |
Unknown | 29 | 36% |
Readers by discipline | Count | As % |
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Chemical Engineering | 1 | 1% |
Psychology | 1 | 1% |
Other | 2 | 2% |
Unknown | 31 | 38% |