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Protein-Protein Interaction Panel Using Mouse Full-Length cDNAs

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Title: Protein-Protein Interaction Panel Using Mouse Full-Length cDNAs
Authors: Suzuki, Harukazu Browse this author
Fukunishi, Yoshifumi Browse this author
Kagawa, Ikuko Browse this author
Saito, Rintaro Browse this author
Oda, Hiroshi Browse this author
Endo, Toshinori Browse this author
Kondo, Shinji Browse this author
Bono, Hidemasa Browse this author
Okazaki, Yasushi Browse this author
Hayashizaki, Yoshihide Browse this author
Issue Date: Oct-2001
Publisher: Cold Spring Harbor Laboratory Press
Journal Title: Genome Research
Volume: 11
Issue: 10
Start Page: 1758
End Page: 1765
Publisher DOI: 10.1101/gr.180101
Abstract: We have developed a novel assay system for systematic analysis of protein–protein interactions (PPIs) that is characteristic of a PCR-mediated rapid sample preparation and a high-throughput assay system based on the mammalian two-hybrid method. Using gene-specific primers, we successfully constructed the assay samples by two rounds of PCR with up to 3.6 kb from the first-round PCR fragments. In the assay system, we designed all the steps to be performed by adding only samples, reagents, and cells into 384-well assay plates using two types of semiautomatic multiple dispensers. The system enabled us examine more than 20,000 assay wells per day. We detected 145 interactions in our pilot study using 3500 samples derived from mouse full-length enriched cDNAs. Analysis of the interaction data showed both several significant interaction clusters and predicted functions of a few uncharacterized proteins. In combination with our comprehensive mouse full-length cDNA clone bank covering a large part of the whole genes, our high-throughput assay system will discover many interactions to facilitate understanding of the function of uncharacterized proteins and the molecular mechanism of crucial biological processes, and also enable completion of a rough draft of the entire PPI panel in certain cell types or tissues of mouse within a short time.
Type: article
Appears in Collections:情報科学院・情報科学研究院 (Graduate School of Information Science and Technology / Faculty of Information Science and Technology) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 遠藤 俊徳

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