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Genes to Cells (2009) 14, 217-225. doi:10.1111/j.1365-2443.2008.01264.x
© 2009 Blackwell Publishing or its licensors

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Localization of gene products using a chromosomally tagged GFP-fusion library in the fission yeast Schizosaccharomyces pombe

Aki Hayashi1a, Ding Da-Qiao1, Chihiro Tsutsumi1, Yuji Chikashige1, Hirohisa Masuda1b, Tokuko Haraguchi1 and Yasushi Hiraoka1,2,*

1 Kobe Advanced ICT Research Center, National Institute of Information and Communications Technology, Kobe 651-2492, Japan
2 Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita 565-0871, Japan

We constructed a library of chromosomally-tagged green fluorescent protein (GFP) fusions in the fission yeast Schizosaccharomyces pombe. This library contains 1058 strains. In each strain, the coding sequence of GFP is integrated at the 3'-end of a particular chromosomal ORF such that the full-length GFP fusion construct is expressed under the control of the original promoter. Integration of the GFP coding sequence at the authentic chromosomal location of each gene was confirmed by PCR. Microscopic screening of these strains detected sufficient levels of GFP signal in 710 strains and allowed assignment of these GFP-fusion gene products with their intracellular localization: 374 proteins were localized in the nucleus, 65 proteins in the nucleolus, 34 proteins at the nuclear periphery, 27 proteins at the plasma membrane and cytoplasmic membranous structures, 24 proteins at the spindle pole body and microtubules, 92 proteins at cytoplasmic structures, and 94 proteins were uniformly distributed throughout the cytoplasm.


Communicated by: Masayuki Yamamoto (The University of Tokyo)

aPresent address: RIKEN, Kobe, Center for Developmental Biology, 2-2-3, Minatozima-minami, Kobe 650-0047, Japan.

bPresent address: Cancer Research UK London Research Institute, Lincoln's Inn Fields Laboratories, London WC2A 3PX, UK.

* Correspondence: hiraoka{at}fbs.osaka-u.ac.jp







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