title: Novel screening system for protein-protein interactions by bimolecular fluorescence complementation in Saccharomyces cerevisiae.
abstract: For high-throughput screening of protein-protein interactions, we have developed a novel yeast screening system using Bimolecular fluorescence complementation (BiFC). Two yeast plasmids, in which genes of heterodimerized peptides LZA and LZB were each fused with those of non-fluorescent half fragments of Kusabira-Green mutant (mKG2), were transformed into a- and α-type yeast, respectively. Mating of them gave a library, which was screened by following green fluorescence resulted from LZA-LZB interaction. The method showed potential ability to detect the positive clones from a model library, in which green-fluorescent and non-fluorescent yeast was mixed in a ratio of 1:675.
authors.affiliation: Department of Biological Science, Osaka Prefecture University, Sakai, Osaka 599-8531, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Kojima
authors.forename: Satoshi
authors.initials: S
authors.lastname: Karasawa
authors.forename: Atsushi
authors.initials: A
authors.lastname: Miyawaki
authors.forename: Takeshi
authors.initials: T
authors.lastname: Tsumuraya
authors.forename: Ikuo
authors.initials: I
authors.lastname: Fujii
mesh_terms: D005434:Flow Cytometry; D005453:Fluorescence; D006801:Humans; D008164:Luminescent Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D010455:Peptides / Q000235:genetics; D025941:Protein Interaction Mapping / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism
chemical_list: D008164:Luminescent Proteins; D010455:Peptides