Identification of proteins that interact with a protein of interest: applications of the yeast two-hybrid system.
The yeast two-hybrid system is a molecular genetic test for protein interaction. Here we describe a step by step procedure to screen for proteins that interact with a protein of interest using the two-hybrid system. This process includes, construction and testing of the bait plasmid, screening a plasmid library for interacting fusion proteins, elimination of false positives and deletion analysis of true positives. This procedure is designed to allow investigators to identify proteins and their encoding cDNAs that have a biologically significant interaction with your protein of interest.
Gietz
R D
RD
Department of Human Genetics, University of Manitoba, Winnipeg, Canada.
Triggs-Raine
B
B
Robbins
A
A
Graham
K C
KC
Woods
R A
RA
D003001:Cloning, Molecular / Q000379:methods*; D005189:False Positive Reactions; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D015723:Gene Library; D017930:Genes, Reporter; D010957:Plasmids / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011485:Protein Binding / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics*; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics
D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase