The bacterial two-hybrid system based on adenylate cyclase reconstitution in Escherichia coli

Methods. 2012 Dec;58(4):325-34. doi: 10.1016/j.ymeth.2012.07.018. Epub 2012 Jul 24.


The bacterial two-hybrid system based on the reconstitution of adenylate cyclase in Escherichia coli (BACTH) was described 14years ago (Karimova, Pidoux, Ullmann, and Ladant, 1998, PNAS, 95:5752). For microbiologists, it is a practical and powerful alternative to the use of the widely spread yeast two-hybrid technology for testing protein-protein interactions. In this review, we aim at giving the reader clear and most importantly simple instructions that should break any reticence to try the technique. Yet, we also add recommendations in the use of the system, related to its specificities. Finally, we expose the advantages and disadvantages of the technique, and review its diverse applications in the literature, which should help in deciding if it is the appropriate method to choose for the case at hand.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Adenylate Cyclase Toxin / biosynthesis*
  • Adenylate Cyclase Toxin / chemistry
  • Adenylate Cyclase Toxin / genetics
  • Catalytic Domain
  • Escherichia coli*
  • Gene Library
  • Genetic Vectors
  • Protein Interaction Domains and Motifs
  • Protein Interaction Mapping / methods
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Two-Hybrid System Techniques*
  • beta-Galactosidase / chemistry


  • Adenylate Cyclase Toxin
  • Recombinant Fusion Proteins
  • beta-Galactosidase