title: Analysis of transcription factor networks using IVV method.
abstract: We have developed a simple and totally in vitro selection procedure based on cell-free cotranslation using a highly stable and efficient in vitro virus (IVV). Cell-free cotranslation of tagged bait and prey proteins is advantageous for the formation of protein complexes and allows high-throughput analysis of protein-protein interactions (PPI) as a result of providing in vitro instead of in vivo preparation of bait proteins. The use of plural selection rounds and a two-step purification of the IVV selection, followed by in vitro post-selection, is advantageous for decreasing false positives. This simple IVV selection system based on cell-free cotranslation is applicable to high-throughput and comprehensive analysis of transcription factor networks.
authors.affiliation: Division of Interactome Medical Sciences, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo, 108-8639, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Ohashi
authors.forename: Shigeo
authors.initials: S
authors.lastname: Fujimori
authors.forename: Naoya
authors.initials: N
authors.lastname: Hirai
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yanagawa
authors.forename: Etsuko
authors.initials: E
authors.lastname: Miyamoto-Sato
mesh_terms: D000818:Animals; D002474:Cell-Free System / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D015723:Gene Library; D006801:Humans; D014176:Protein Biosynthesis; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; D014157:Transcription Factors