High-throughput yeast two-hybrid assays for large-scale protein interaction mapping.
Protein-protein interactions play fundamental roles in many biological processes. Hence, protein interaction mapping is becoming a well-established functional genomics approach to generate functional annotations for predicted proteins that so far have remained uncharacterized. The yeast two-hybrid system is currently one of the most standardized protein interaction mapping techniques. Here, we describe the protocols for a semiautomated, high-throughput, Gal4-based yeast two-hybrid system.
Walhout
A J
AJ
Dana-Farber Cancer Institute and Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.
Vidal
M
M
D003470:Culture Media; D004274:DNA, Recombinant / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D017930:Genes, Reporter / Q000235:genetics; D066298:In Vitro Techniques; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014170:Transformation, Genetic; D020798:Two-Hybrid System Techniques
D003470:Culture Media; D004274:DNA, Recombinant; D005656:Fungal Proteins; D011506:Proteins