Novel fluorescence labeling and high-throughput assay technologies for in vitro analysis of protein interactions

Nobuhide Doi, Hideaki Takashima, Masataka Kinjo, Kyoko Sakata, Yuko Kawahashi, Yuko Oishi, Rieko Oyama, Etsuko Miyamoto-Sato, Tatsuya Sawasaki, Yaeta Endo, Hiroshi Yanagawa

研究成果: Article査読

70 被引用数 (Scopus)

抄録

We developed and tested a simple method for fluorescence labeling and interaction analysis of proteins based on a highly efficient in vitro translation system combined with high-throughput technologies such as microarrays and fluorescence cross-correlation spectroscopy (FCCS). By use of puromycin analogs linked to various fluorophores through a deoxycytidylic acid linker, a single fluorophore can be efficiently incorporated into a protein at the carboxyl terminus during in vitro translation. We confirmed that the resulting fluorescently labeled proteins are useful for probing protein-protein and protein-DNA interactions by means of pulldown assay, DNA microarrays, and FCCS in model experiments. These fluorescence assay systems can be easily extended to highly parallel analysis of protein interactions in studies of functional genomics.

本文言語English
ページ(範囲)487-492
ページ数6
ジャーナルGenome Research
12
3
DOI
出版ステータスPublished - 2002

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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