biostudies-literatureUnknown46(3)Yang FProtein-protein interaction (PPI) network maintains proper function of all organisms. Simple high-throughput technologies are desperately needed to delineate the landscape of PPI networks. While recent state-of-the-art yeast two-hybrid (Y2H) systems improved screening efficiency, either individual colony isolation, library preparation arrays, gene barcoding or massive sequencing are still required. Here, we developed a recombination-based 'library vs library' Y2H system (RLL-Y2H), by which multi-library screening can be accomplished in a single pool without any individual treatment. This system is based on the phiC31 integrase-mediated integration between bait and prey plasmids. The integrated fragments were digested by MmeI and subjected to deep sequencing to decode the interaction matrixNucleic acids researche17https://www.ebi.ac.uk/biostudies/studies/S-EPMC5815087biostudies-literatureDevelopment and application of a recombination-based library versus library high- throughput yeast two-hybrid (RLL-Y2H) screening system.PMC5815087Zhou MHabib ZCao GWang DFu ZZhong WHan YYao QChen HXiao SYang FRahman KZhu CLin DLei YTao RChen XXu WTyagi RWu XYu YfalseDevelopment and application of a recombination-based library versus library high- throughput yeast two-hybrid (RLL-Y2H) screening system.Protein-protein interaction (PPI) network maintains proper function of all organisms. Simple high-throughput technologies are desperately needed to delineate the landscape of PPI networks. While recent state-of-the-art yeast two-hybrid (Y2H) systems improved screening efficiency, either individual colony isolation, library preparation arrays, gene barcoding or massive sequencing are still required. Here, we developed a recombination-based 'library vs library' Y2H system (RLL-Y2H), by which multi-library screening can be accomplished in a single pool without any individual treatment. This system is based on the phiC31 integrase-mediated integration between bait and prey plasmids. The integrated fragments were digested by MmeI and subjected to deep sequencing to decode the interaction matrix2018-01-01T00:00:00Z2018 Feb2026-04-28T22:42:00.987Z2019-03-26T23:03:30ZS-EPMC58150872916564610.1093/nar/gkx1173