A bacterial/yeast merged two-hybrid system: protocol for yeast screening with single or parallel baits.
The yeast two-hybrid system is a useful tool for identifying new protein-protein interactions, and for the dissection of previously identified interactions. An important issue in protein-interaction studies is frequently that of determining whether a protein associates specifically with one protein or domain of interest, or has a more promiscuous interaction profile. To help address this issue, the authors have created a new two-hybrid system, which can be used either in bacteria or in yeast to counterscreen against "decoy" baits in parallel with a primary screen, hence improving the power and specificity of the method. Protocols of this system for use in yeast are provided; a companion article, Serebriiski et al., describes alternative use of this system in bacteria.
Tikhmyanova
Nadezhda Y
NY
Fox Chase Cancer Center, Philadelphia, PA, USA.
Izumchenko
Eugene A
EA
Serebriiskii
Ilya G
IG
Golemis
Erica A
EA
D000595:Amino Acid Sequence; D001419:Bacteria / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D015723:Gene Library; D017930:Genes, Reporter; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010641:Phenotype; D010957:Plasmids / Q000235:genetics; D025941:Protein Interaction Mapping / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic; D020798:Two-Hybrid System Techniques
D017931:DNA Primers; D004274:DNA, Recombinant