ArticleGenome research2003
A strategy for constructing large protein interaction maps using the yeast two-hybrid system: regulated expression arrays and two-phase mating.
Article in Genome research, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
- The protein interactome of the citrus Huanglongbing pathogen Candidatus Liberibacter asiaticus.Nature communications · 2023Article
- Genome-wide inference of the Camponotus floridanus protein-protein interaction network using homologous mapping and interacting domain profile pairs.Scientific reports · 2020Article
- Article
- Targeting mammalian organelles with internalizing phage (iPhage) libraries.Nature protocols · 2013Article
- Identification of new protein interactions between dengue fever virus and its hosts, human and mosquito.PloS one · 2013Article
- Diversity in genetic in vivo methods for protein-protein interaction studies: from the yeast two-hybrid system to the mammalian split-luciferase system.Microbiology and molecular biology reviews : MMBR · 2012Review
- Article
- High throughput flow cytometry based yeast two-hybrid array approach for large-scale analysis of protein-protein interactions.Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2012Article
- A dynamic DUO of regulatory proteins coordinates gamete specification and germ cell mitosis in the angiosperm male germline.Plant signaling & behavior · 2009Article
- Udu deficiency activates DNA damage checkpoint.Molecular biology of the cell · 2009Article
- Shifted Transversal Design smart-pooling for high coverage interactome mapping.Genome research · 2009Article
- Arabidopsis DUO POLLEN3 is a key regulator of male germline development and embryogenesis.The Plant cell · 2009Article
- Cost-effective strategies for completing the interactome.Nature methods · 2009Article
- A yeast two-hybrid smart-pool-array system for protein-interaction mapping.Nature methods · 2007Article
- A proteome-wide protein interaction map for Campylobacter jejuni.Genome biology · 2007Article
- The protein network of bacterial motility.Molecular systems biology · 2007Article
- Display technologies: application for the discovery of drug and gene delivery agents.Advanced drug delivery reviews · 2006Review
- A pooling-deconvolution strategy for biological network elucidation.Nature methods · 2006Article
- Hematopoietic stem cells: expression profiling and beyond.Stem cell reviews · 2006Review
- Novel mobilizable prokaryotic two-hybrid system vectors for high-throughput protein interaction mapping in Escherichia coli by bacterial conjugation.Nucleic acids research · 2005Article
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5 authors.
Funding
Abstract
Maps representing the binary interactions among proteins have become valuable tools for understanding how proteins work together to mediate biological processes. One of the most effective methods for detecting biologically important protein interactions has been the yeast two-hybrid system. Here we present an efficient two-hybrid strategy to facilitate construction of protein interaction maps on a genome-wide scale. The strategy begins with two arrays of yeast expressing known proteins fused to either a DNA binding domain (BD), or a transcription activation domain (AD). The fusion proteins are conditionally expressed using regulated promoters that can be repressed during construction and amplification of the yeast arrays. Interaction assays are conducted in two phases. In the first phase, small pools of AD strains are mated with the array of BD strains. In the second phase, individual BD strains are mated with appropriate subsets of the AD array corresponding to positive pools in the first phase. This strategy has several advantages over previously described approaches, including the ability to detect interactions with proteins that inhibit yeast growth or that activate transcription as BD fusions. Moreover, by minimizing the number of mating operations and sequencing reactions needed to test large sets of binary interactions, this strategy is more efficient than either matrix or library screening approaches. We also present a three-dimensional pooling scheme to further increase the efficiency of large-scale two-hybrid analyses.
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