Use of cDNA Tiling Arrays for Identifying Protein Interactions Selected by In Vitro Display Technologies

Kenichi Horisawa, Nobuhide Doi, Hiroshi Yanagawa

研究成果: Article

7 引用 (Scopus)

抜粋

In vitro display technologies such as mRNA display are powerful screening tools for protein interaction analysis, but the final cloning and sequencing processes represent a bottleneck, resulting in many false negatives. Here we describe an application of tiling array technology to identify specifically binding proteins selected with the in vitro virus (IVV) mRNA display technology. We constructed transcription-factor tiling (TFT) arrays containing ~1,600 open reading frame sequences of known and predicted mouse transcription-regulatory factors (334,372 oligonucleotides, 50-mer in length) to analyze cDNA fragments from mRNA-display screening for Jun-associated proteins. The use of the TFT arrays greatly increased the coverage of known Jun-interactors to 28% (from 14% with the cloning and sequencing approach), without reducing the accuracy (~75%). This method could detect even targets with extremely low expression levels (less than a single mRNA copy per cell in whole brain tissue). This highly sensitive and reliable method should be useful for high-throughput protein interaction analysis on a genome-wide scale.

元の言語English
記事番号e0001646
ジャーナルPloS one
3
発行部数2
DOI
出版物ステータスPublished - 2008

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

フィンガープリント Use of cDNA Tiling Arrays for Identifying Protein Interactions Selected by In Vitro Display Technologies' の研究トピックを掘り下げます。これらはともに一意のフィンガープリントを構成します。

  • これを引用