Membrane and Nuclear Yeast Two-Hybrid Systems

The yeast two-hybrid (Y2H) system is an effective means of detecting protein-protein interactions through the activation of reporter gene expression. This system has also displayed several useful applications, including rapidly revealing unexpected interactors of known proteins, detecting regions that take part in protein-protein interactions, and characterizing the orchestration of protein interactions in metabolic pathways. The Y2H system has been widely utilized in the study of plant virology to investigate interactions between viral proteins and host, vector, or viral proteins. This technology has been successful in enhancing the understanding of viral structure and assembly, replication, viral gene expression and regulation, viral movement, pathogenicity, and the network of viral proteins. Here, we provide the methods to uncover novel interactors of viral proteins via screening cDNA libraries of plant host and insect vectors, using nuclear and membrane Y2H systems, respectively.