Abstract
The yeast two-hybrid system enables investigation of protein-protein interactions and provides a powerful functional expression cloning system by combining molecular engineering, reporter genes and yeast genetics. The concept of a two-hybrid interaction system was first published in 1989. Since then the basic premise of the yeast two-hybrid system has undergone modifications, variations and extensions. These adaptations enable the application of yeast two-hybrid methods to a variety of diverse scientific questions, well beyond that described for the system ten years ago. The scientific and patent literature initially described the use of two-hybrid systems to study specific protein-protein interactions, then progressed to modifications and adaptations on two-hybrid methods, and leads to descriptions on the extended utility and expansion of yeast two-hybrid techniques with which to address complex biological questions. Improvements and modifications in vectors, reporter gene cassettes and yeast strains, which comprise the fundamental elements of yeast two-hybrid, have contributed to its evolution. The utility of the basic two-hybrid technique has been expanded to study interactions between multiple proteins, nucleotides and proteins, as well as various chemical linkers and proteins. The versatility and inherent flexibility of a yeast two-hybrid based approach enables this method, and extensions thereof, to serve as potential platform technologies for target identification and characterisation, genome mapping, functional genomics, compound screening, small molecule identification and drug discovery. As such, yeast two-hybrid based technologies are increasingly embedded within fully integrated biotechnology and pharmaceutical research programmes geared toward drug discovery.
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