Nature Biotechnology 24, 841 - 847 (2006)
Published online: 25 June 2006; | doi:10.1038/nbt1222
There is a Corrigendum associated with this Letter. Please see the PDF for details.ORFeome cloning and global analysis of protein localization in the fission yeast Schizosaccharomyces pombeAkihisa Matsuyama1, 2, 6, Ritsuko Arai1, 2, 6, Yoko Yashiroda1, 2, 6, Atsuko Shirai1, 2, 3, Ayako Kamata1, 2, 3, Shigeko Sekido1, Yumiko Kobayashi1, Atsushi Hashimoto1, Makiko Hamamoto1, 5, Yasushi Hiraoka2, 4, Sueharu Horinouchi3
& Minoru Yoshida1, 2, 31
Chemical Genetics Laboratory, RIKEN, Wako, Saitama
351-0198, Japan. 2
CREST Research Project, JST, Kawaguchi, Saitama
332-0012, Japan. 3
Department of Biotechnology, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo
113-8657, Japan. 4
Kansai Advanced Research Center, National Institute of Information and Communications Technology, Kobe, Hyogo
651-2492, Japan. 5
Present address: School of Agriculture, Meiji University, Tama-ku, Kanagawa
214-8571, Japan. 6
These authors contributed equally to this work.
Correspondence should be addressed to Minoru Yoshida yoshidam@riken.jp Cloning of the entire set of an organism's protein-coding open reading frames (ORFs), or 'ORFeome', is a means of connecting the genome to downstream 'omics' applications. Here we report a proteome-scale study of the fission yeast Schizosaccharomyces pombe based on cloning of the ORFeome. Taking advantage of a recombination-based cloning system, we obtained 4,910 ORFs in a form that is readily usable in various analyses. First, we evaluated ORF prediction in the fission yeast genome project by expressing each ORF tagged at the 3' terminus. Next, we determined the localization of 4,431 proteins, corresponding to  90% of the fission yeast proteome, by tagging each ORF with the yellow fluorescent protein. Furthermore, using leptomycin B, an inhibitor of the nuclear export protein Crm1, we identified 285 proteins whose localization is regulated by Crm1.
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