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 |
| 巻 | 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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All Science Journal Classification (ASJC) codes
- Biochemistry
- Molecular Biology
これを引用
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
}
TY - JOUR
T1 - High-throughput fluorescence labelling of full-length cDNA products based on a reconstituted translation system
AU - Kawahashi, Yuko
AU - Doi, Nobuhide
AU - Oishi, Yo
AU - Tsuda, Chizuru
AU - Takashima, Hideaki
AU - Baba, Tomoya
AU - Mori, Hirotada
AU - Ito, Takashi
AU - Yanagawa, Hiroshi
PY - 2007/1/1
Y1 - 2007/1/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=34147219548&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34147219548&partnerID=8YFLogxK
U2 - 10.1093/jb/mvm003
DO - 10.1093/jb/mvm003
M3 - Article
C2 - 17148548
AN - SCOPUS:34147219548
VL - 141
SP - 19
EP - 24
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 1
ER -