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Book Sections Year : 2017

Protein-Protein Interaction: Bacterial Two-Hybrid


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.
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pasteur-02879054 , version 1 (23-06-2020)



Gouzel Karimova, Emilie Gauliard, Marilyne Davi, Scot Ouellette, Daniel Ladant. Protein-Protein Interaction: Bacterial Two-Hybrid. Laure Journet; Eric Cascales. Bacterial Protein Secretion Systems, 1615, Humana Press; Springer Science, pp.159-176, 2017, Methods in Molecular Biology, 978-1-4939-7033-9. ⟨10.1007/978-1-4939-7033-9_13⟩. ⟨pasteur-02879054⟩
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