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Book sections
Protein-Protein Interaction: Bacterial Two-Hybrid
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.
https://hal-pasteur.archives-ouvertes.fr/pasteur-02879054
Contributor : Gouzel Karimova Connect in order to contact the contributor
Submitted on : Tuesday, June 23, 2020 - 2:48:53 PM
Last modification on : Thursday, November 25, 2021 - 4:00:02 PM
Contributor : Gouzel Karimova Connect in order to contact the contributor
Submitted on : Tuesday, June 23, 2020 - 2:48:53 PM
Last modification on : Thursday, November 25, 2021 - 4:00:02 PM