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Localization studies using GFP technology and reference dyesGreen-fluorescent-protein (GFP) fusions are indispensable tools for studying the distribution and dynamics of proteins and organelles in vivo. We use GFP and variants thereof to determine the spatial subcellular localization of proteins involved in the lipid metabolism of the yeast. In this respect, we study the expression of GFP tagged proteins in single cells, in larger cell populations (quantitative analysis) or globally using commercially available or in-house produced strain libraries (1,2). Generated localization data are partly public (YPL, Yeast Protein Localization Database (3,4)) and can be accessed via Saccharomyces cerevisiae genome database (SGD, Stanford University). In addition to GFP fusions we apply commerically available reference dyes for studying interactions of proteins and organelles.
1. Natter K, Leitner P, Faschinger A, Wolinski H, McCraith S, Fields S, 2. Kohlwein SD, The spatial organization of lipid synthesis in the yeast Saccharomyces cerevisiae derived from large scale green fluorescent protein tagging and high resolution microscopy. Mol Cell Proteomics. 2005; 4(5):662-72.
2. Kohlwein SD. The beauty of the yeast: live cell microscopy at the limits of optical resolution. Microsc Res Tech. 2000; 51, 511-29.
3. Habeler G, Natter K, Thallinger GG, Crawford ME, Kohlwein SD, Trajanoski Z., YPL.db: the Yeast Protein Localization database. Nucleic Acids Res. 2002 ;30(1):80-3.
4. Kals M, Natter K, Thallinger GG, Trajanoski Z, Kohlwein SD, YPL.db2: the Yeast Protein Localization database, version 2.0. Yeast. 2005; 22(3):213-8.
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