Methods for immunoelectron microscopic and fine structural analysis of synaptonemal complexes and nodules in yeast |
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Authors: | Karin Schmekel |
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Institution: | (1) Department of Molecular Genome Research, Stockholm University, 10691 Stockholm, Sweden, SE |
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Abstract: | Several gene products involved in meiotic chromosome pairing and recombination in yeast have been identified in recent years.
Two nuclear structures play key roles in the meiotic processes: the synaptonemal complex (SC), which is essential for the
pairing of the chromosomes, and the recombination nodules (RNs), which mark the sites of recombination. Good morphological
representation of the yeast SC and RNs is needed in order to show structural changes caused by specific mutations in protein-coding
genes and for fine localization of proteins using immunoelectron microscopy (immuno-EM). This paper presents a newly developed
preparation method for EM and immuno-EM that allows analysis of fine structural details and localization of proteins in the
SC and RNs in yeast. Structural components of the SC are clearly seen and appear strikingly similar to those in the SC in
other organisms. Antibodies against the SC protein Zip1, a transverse filament protein, label the central region of the SC
strongly and specifically as expected. The improved method will be an important tool in high-resolution determination of the
location of proteins in the meiotic yeast nucleus.
Received: 9 March 1999; in revised form: 1 September 1999 / Accepted: 22 September 1999 |
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