High-throughput fluorescence labelling of full-length cDNA products based on a reconstituted translation system

Yuko Kawahashi, Nobuhide Doi, Yo Oishi, Chizuru Tsuda, Hideaki Takashima, Tomoya Baba, Hirotada Mori, Takashi Ito, Hiroshi Yanagawa

研究成果: Article

4 引用 (Scopus)

抜粋

Although recent advances in fluorescence-based technologies, such as protein microarrays, have made it possible to analyse more than 10,000 proteins at once, there is a bottleneck in the step of preparation of large numbers of fluorescently labelled proteins for the comprehensive analysis of protein-protein interactions. Here we describe two independent methods for high-throughput fluorescence-labelling of full-length cDNA products at their C-termini using a reconstituted translation system containing fluorescent puromycin. For the first method, release factor-free systems were used. For the second method, stop codons were excluded from cDNAs by using a common mismatch primer in mutagenic PCR. These methods yielded large numbers of labelled proteins from cDNA sets of various organisms, such as mouse, yeast and Escherichia coli.

元の言語English
ページ(範囲)19-24
ページ数6
ジャーナルJournal of biochemistry
141
発行部数1
DOI
出版物ステータスPublished - 2007 1

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

フィンガープリント High-throughput fluorescence labelling of full-length cDNA products based on a reconstituted translation system' 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

    Kawahashi, Y., Doi, N., Oishi, Y., Tsuda, C., Takashima, H., Baba, T., Mori, H., Ito, T., & Yanagawa, H. (2007). High-throughput fluorescence labelling of full-length cDNA products based on a reconstituted translation system. Journal of biochemistry, 141(1), 19-24. https://doi.org/10.1093/jb/mvm003