Localization of gene products using a chromosomally tagged GFP‐fusion library in the fission yeast Schizosaccharomyces pombe
Open Access
- 12 January 2009
- journal article
- research article
- Published by Wiley in Genes to Cells
- Vol. 14 (2), 217-225
- https://doi.org/10.1111/j.1365-2443.2008.01264.x
Abstract
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.Keywords
This publication has 20 references indexed in Scilit:
- Reconstruction of the Kinetochore during Meiosis in Fission Yeast Schizosaccharomyces pombeMolecular Biology of the Cell, 2006
- ORFeome cloning and global analysis of protein localization in the fission yeast Schizosaccharomyces pombeNature Biotechnology, 2006
- Global analysis of protein localization in budding yeastNature, 2003
- Revealing modular organization in the yeast transcriptional networkNature Genetics, 2002
- Subcellular localization of the yeast proteomeGenes & Development, 2002
- Systematic subcellular localization of novel proteins identified by large‐scale cDNA sequencingEMBO Reports, 2000
- Multiple-color Fluorescence Imaging of Chromosomes and Microtubules in Living Cells.Cell Structure and Function, 1999
- Identification of fission yeast nuclear markers using random polypeptide fusions with green fluorescent proteinProceedings of the National Academy of Sciences of the United States of America, 1996
- Minimal extent of homology required for completion of meiotic recombination inSaccharomyces cerevisiaeDevelopmental Genetics, 1992
- [56] Molecular genetic analysis of fission yeast Schizosaccharomyces pombeMethods in enzymology, 1991