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Review
. 2009:557:65-76.
doi: 10.1007/978-1-59745-527-5_6.

Genetic analysis of meiotic recombination in Schizosaccharomyces pombe

Gerald R Smith  1
Affiliations
Review

Genetic analysis of meiotic recombination in Schizosaccharomyces pombe

Gerald R Smith. Methods Mol Biol. 2009.

Abstract

The fission yeast Schizosaccharomyces pombe is well-suited for studying meiotic recombination. Methods are described here for culturing S. pombe and for genetic assays ofintragenic recombination (gene conversion), intergenic recombination (crossing-over), and spore viability. Both random spore and tetrad analyses are described.

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Figures

Fig. 1
Fig. 1
Asci from h90 meiosis. Cells of an h90 strain were washed and spotted on a supplemented SPA plate, which was incubated for 3 d at 25°. A sample of the cell-ascus mixture was observed under a differential interference contrast microscope and photographed. (Left) The linear ascus (A) has the appearance of an azygotic ascus, whereas the curved ascus (B) has that of a zygotic ascus. A zygote (Z), an unmated cell (C), and a spore (S) from spontaneous lysis of an ascus are also visible in this rare field. (Right) A more typical field of asci, one of which (Z) is immature.
Fig. 2
Fig. 2
Assembly of Petri dish with acetone-soaked paper for killing vegetative cells. A replicating velveteen (a) is stretched over the elevated plastic block (b) and held in place with the clear plastic ring (c). A master plate with colonies is replicated onto the velveteen. Acetone is pipetted onto the filter paper (d) held in the glass Petri dish (e) with an internal plastic ring (f). This assembly is then inverted over the velveteen. After exposure to the acetone vapors, the cell-ascus mixtures on the velveteen are replicated onto a fresh YEA plate.
Fig. 3
Fig. 3
Grid (full size) for picking colonies for replication to test media. A fresh YEA plate is centered over this grid. Cells are transferred from colonies onto the YEA plate, one over each grid mark. After incubation, this plate is replicated to test media.
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References

    1. Davis L, Smith GR. Non-random homolog segregation at meiosis I in Schizosaccharomyces pombe mutants lacking recombination. Genetics. 2003;163:857–874. - PMC - PubMed
    1. Cromie GA, Smith GR. In: Genome Dynamics and Stability. Egel R, editor. Springer-Verlag; Berlin: in press .
    1. Wood V, Gwilliam R, Rajandream MA, Lyne M, Lyne R, Stewart A, Sgouros J, Peat N, Hayles J, Baker S, Basham D, et al. The genome sequence of Schizosaccharomyces pombe. Nature. 2002;415:871–880. - PubMed
    1. Moreno S, Klar AJS, Nurse P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 1991;194:795–823. - PubMed
    1. Egel R, editor. The molecular biology of Schizosaccharomyces pombe. Springer-Verlag; Berlin: 2004.

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