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: ジャーナルへの寄稿記事

3 引用 (Scopus)

抄録

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.

元の言語英語
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
出版物ステータス出版済み - 1 1 2007

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Labeling
Complementary DNA
Fluorescence
Throughput
Proteins
Puromycin
Protein Array Analysis
Terminator Codon
Microarrays
Yeast
Escherichia coli
Yeasts
Technology
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

これを引用

Kawahashi, Y., Doi, N., Oishi, Y., Tsuda, C., Takashima, H., Baba, 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

High-throughput fluorescence labelling of full-length cDNA products based on a reconstituted translation system. / Kawahashi, Yuko; Doi, Nobuhide; Oishi, Yo; Tsuda, Chizuru; Takashima, Hideaki; Baba, Tomoya; Mori, Hirotada; Ito, Takashi; Yanagawa, Hiroshi.

:: Journal of biochemistry, 巻 141, 番号 1, 01.01.2007, p. 19-24.

研究成果: ジャーナルへの寄稿記事

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, 巻. 141, 番号 1, pp. 19-24. https://doi.org/10.1093/jb/mvm003
Kawahashi, Yuko ; Doi, Nobuhide ; Oishi, Yo ; Tsuda, Chizuru ; Takashima, Hideaki ; Baba, Tomoya ; Mori, Hirotada ; Ito, Takashi ; Yanagawa, Hiroshi. / High-throughput fluorescence labelling of full-length cDNA products based on a reconstituted translation system. :: Journal of biochemistry. 2007 ; 巻 141, 番号 1. pp. 19-24.
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