@article{Miyamoto-Sato01052005,
author = {Miyamoto-Sato, Etsuko and Ishizaka, Masamichi and Horisawa, Kenichi and Tateyama, Seiji and Takashima, Hideaki and Fuse, Shinichiro and Sue, Kaori and Hirai, Naoya and Masuoka, Kazuyo and Yanagawa, Hiroshi}, 
title = {Cell-free cotranslation and selection using in vitro virus for high-throughput analysis of protein–protein interactions and complexes},
volume = {15}, 
number = {5}, 
pages = {710-717}, 
year = {2005}, 
doi = {10.1101/gr.3510505}, 
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. In a single experiment using bait Fos, more than 10 interactors, including not only direct, but also indirect interactions, were enriched. Further, previously unidentified proteins containing novel leucine zipper (L-ZIP) motifs with minimal binding sites identified by sequence alignment as functional elements were detected as a result of using a randomly primed cDNA library. Thus, we consider that this simple IVV selection system based on cell-free cotranslation could be applicable to high-throughput and comprehensive analysis of PPI and complexes in large-scale settings involving parallel bait proteins.}, 
URL = {http://genome.cshlp.org/content/15/5/710.abstract}, 
eprint = {http://genome.cshlp.org/content/15/5/710.full.pdf+html}, 
journal = {Genome Research} 
}