Yeast two-hybrid screening.
Yeast two-hybrid screening represents a sensitive in vivo method for the identification and analysis of protein-protein interactions. The principle is based on the ability of a separate DNA-binding domain (DNA-BD) and activation domain (AD) to reconstitute a functional transactivator when brought into proximity. In the MATCHMAKER yeast two-hybrid system, a bait protein is expressed as a fusion to the GAL4 DNA-BD, whereas the prey protein is expressed as a fusion to the GAL4 AD. When a bait and a prey protein interact, the DNA-BD and AD form a functional transactivator, resulting in activation of reporter gene expression in yeast reporter strains. The method described in this chapter can be used to identify novel protein interactions, analyze protein-protein interactions between two known proteins, as well as dissect interacting protein domains.
Maple
Jodi
J
Department of Mathematics and Natural Sciences, University of Stavanger, Norway.
Møller
Simon G
SG
D001483:Base Sequence; D004274:DNA, Recombinant / Q000235:genetics; D004268:DNA-Binding Proteins; D017930:Genes, Reporter; D005822:Genetic Vectors; D025941:Protein Interaction Mapping; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic; D020798:Two-Hybrid System Techniques
D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors