Chapter 27
Yeast FISH: Delineation of Chromosomal Targets
in Vegetative and Meiotic Yeast Cells
HARRY SCHERTHAN * and EDGAR TRELLES-STICKEN *
Introduction
Fluorescence in situ hybridization provides an effective means to delineate
chromosomes and chromosomal regions of interest, during all stages of
the cell cycle, irrespective of chromatin composition and compaction. This
property makes FISH particularly useful for studying chromosome behavior in species with minute genomes and/or ill-defined metaphase chromosomes. Genetic model organisms like fission and budding yeast display
such cytological characteristics. Consequently, chromosome painting and
locus-specific yeast FISH (Scherthan et al. 1992, 1994, Uzawa et al. 1992,
Guacci et al. 1994, Weiner and Kleckner 1994, Gotta et al. 1996) has become an indispensable tool in the analysis of chromosome dynamics and
organization in wild-type and mutant yeast strains.
Principles and applications
As compared to human and other higher eukaryotes, the bakers yeast Saccharomyces cerevisiae (n=16) challenges classical cytology, since metaphase occurs without nuclear envelope breakdown, highly condensed
chromosomes are absent and yeast nuclei lack a lamina which makes
them vulnerable to distortions during cell wall removal and isolation procedures. Conventional cytogenetic analysis of yeast mitosis, therefore,
only reveals a morphological change of the nuclear/chromosome mass
which is seen to elongate, bifurcate and eventually to separate during ana~ Harry Scherthan, Edgar Trelles-Sticken, University of Kaiserslautern, Dept. of
Human Biology and Human Genetics, Erwin-Schroedingerstr., Kaiserslautern, 67663,
Germany (phone +49-631-205-2107; fax +49-631-205-2878; e-mail scherth@uni.de)
* present address: Max-Planck-Inst. for Molecular Genetics, Ihnestr. 73, Berlin,
0-14195, Germany.
PROTOCOL
Yeast FISH: Delineation of Chromosomal Targets
in Vegetative and Meiotic Yeast Cells
HARRY SCHERTHAN * and EDGAR TRELLES-STICKEN *
Introduction
Fluorescence in situ hybridization provides an effective means to delineate
chromosomes and chromosomal regions of interest, during all stages of
the cell cycle, irrespective of chromatin composition and compaction. This
property makes FISH particularly useful for studying chromosome behavior in species with minute genomes and/or ill-defined metaphase chromosomes. Genetic model organisms like fission and budding yeast display
such cytological characteristics. Consequently, chromosome painting and
locus-specific yeast FISH (Scherthan et al. 1992, 1994, Uzawa et al. 1992,
Guacci et al. 1994, Weiner and Kleckner 1994, Gotta et al. 1996) has become an indispensable tool in the analysis of chromosome dynamics and
organization in wild-type and mutant yeast strains.
Principles and applications
As compared to human and other higher eukaryotes, the bakers yeast Saccharomyces cerevisiae (n=16) challenges classical cytology, since metaphase occurs without nuclear envelope breakdown, highly condensed
chromosomes are absent and yeast nuclei lack a lamina which makes
them vulnerable to distortions during cell wall removal and isolation procedures. Conventional cytogenetic analysis of yeast mitosis, therefore,
only reveals a morphological change of the nuclear/chromosome mass
which is seen to elongate, bifurcate and eventually to separate during ana~ Harry Scherthan, Edgar Trelles-Sticken, University of Kaiserslautern, Dept. of
Human Biology and Human Genetics, Erwin-Schroedingerstr., Kaiserslautern, 67663,
Germany (phone +49-631-205-2107; fax +49-631-205-2878; e-mail scherth@uni.de)
* present address: Max-Planck-Inst. for Molecular Genetics, Ihnestr. 73, Berlin,
0-14195, Germany.
PROTOCOL