Abstract
Tetrad analysis of most Saccharomyces strains used in beer production is impossible due to a low yield of viable spores. The present paper describes the use of single chromosome transfer in the genetic analysis of a brewer’s yeast.
The technique employs thekar1 mutation, which reduces karyogamy after conjugation. In rare caseskarl×KAR crosses yield progeny resulting from the transfer of one chromosome or a limited number of chromosomes from a nucleus of one parent to one of the other. S. cerevisiae strains with an extra S. carlsbergensis chromosome III have thus been isolated from crosses between spore derived clones of the brewing strain and haploidkar1 S. cerevisiae strains carrying several auxotrophic markers. When the disomics were crossed to other haploid S. cerevisiae strains a normal spore viability was obtained, allowing tetrad analysis.
High functional homology was found between the transferred S. carlsbergensis chromosome and chromosome III of S. cerevisiae. All genes essential for viability on the latter are represented on the former as are alsoHIS4, LEU2, MAT andTHR4. Despite the functional homology, the transferred chromosome had a structure that was substantially different from that of standard S. cerevisiae strains. It did not recombine with S. cerevisiae chromosomes III, except in a certain region, where recombination was normal. Furthermore, restriction endonuclease analysis showed that the variant chromosome has a nucleotide sequence in theHIS4 region different from that of S. cerevisiae. The S. carlsbergensis brewing strain is heterozygous for this sequence variation, containing also aHIS4 region with a sequence identical or close to that of S. cerevisiae.
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Abbreviations
- NM:
-
non-mater
- NPD:
-
non-parental ditype
- PD:
-
parental ditype
- TT:
-
tetratype
- YPD:
-
yeast extract peptone dextrose
References
Anderson, E. &P. A. Martin: The sporulation and mating of brewing yeasts. J. Inst. Brew. 81, 242–247 (1975)
Conde, J. &G. R. Fink: A mutant ofSaccharomyces cerevisiae defective for nuclear fusion. Proc. Nat. Acad. Sci USA 73, 3651–3655 (1976)
Fowell, R. R.: Sporulation and hybridization of yeasts. In: The Yeasts. A. H. Rose & J. Harrison, eds., Academic Press, London and New York, Vol. I, pp 303–383 (1969)
Gjermansen, C. &P. Sigsgaard: Construction of a hybrid brewing strain of Saccharomyces carlsbergensis by mating of meiotic segregants. Carlsberg Res. Commun. 46, 1–11 (1981)
Harashima, S. &Y. Oshima: Mapping of the homothallic genes,HMα andHMa in Saccharomyces yeasts. Genetics 84, 437–451 (1976)
Hawthorne, D. C.: A deletion in yeast and its bearing on the structure of the mating type locus. Genetics 48, 1727–1729 (1963)
Hawthorne, D. C. &R. K. Mortimer: Chromosome mapping in Saccharomyces: Centromere-linked genes. Genetics 45, 1085–1110 (1960)
Holmberg, S., M. C. Kielland-Brandt, T. Nilsson-Tillgren &J. G. L. Petersen: Molecular characterization of threehis4 deletion mutants in Saccharomyces cerevisiae. Carlsberg Res. Commun. 44, 283–288 (1979)
Holmberg, S., J. G. L. Petersen, T. Nilsson-Tillgren &M. C. Kielland-Brandt: Molecular characterization of a Saccharomyces plasmid containing theHIS4 gene. Carlsberg Res. Commun. 44, 269–282 (1979)
Kielland-Brandt, M. C., T. Nilsson-Tillgren, S. Holmberg, J. G. L. Petersen &B. A. Svenningsen: Transformation of yeast without the use of foreign DNA. Carlsberg Res. Commun. 44, 77–87 (1979)
Mortimer, R. K. &D. C. Hawthorne: Genetic mapping in yeast. Meth. Cell Biol. 11, 221–233 (1975)
Nilsson-Tillgren, T., J. G. L. Petersen, S. Holmberg &M. C. Kielland-Brandt: Stabilization by chromosomal integration ofHIS4 from an unstable yeast transformant. Abst. in The molecular Biology of Yeast. Cold Spring Harbor Laboratory, New York p. 125 (1979)
Nilsson-Tillgren, T., J. G. L. Petersen, S. Holmberg &M. C. Kielland-Brandt: Transfer of chromosome III duringkar mediated cytoduction in yeast. Carlsberg Res. Commun. 45, 113–117 (1980)
Southern, E. M.: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98, 503–517 (1975)
Strathern, J. N., C. S. Newlon, I. Herskowitz &J. B. Hicks: Isolation of a circular derivative of yeast chromosome III: Implications for the mechanism of mating type interconversion. Cell 18, 309–319 (1979)
Tabak, H. F. &R. A. Flavell: A method for the recovery of DNA from agarose gels. Nucl. Acids Res. 5, 2321–2332 (1978)
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Nilsson-Tillgren, T., Gjermansen, C., Kielland-Brandt, M.C. et al. Genetic differences between Saccharomyces carlsbergensis and S. cerevisiae. Analysis of chromosome III by single chromosome transfer. Carlsberg Res. Commun. 46, 65–76 (1981). https://doi.org/10.1007/BF02906199
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DOI: https://doi.org/10.1007/BF02906199