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Molecular probes for the detection of Kluyveromyces marxianus chromosomal DNA in electrophoretic karyotypes of intergeneric protoplast fusion products

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Abstract

Random genomic DNA fragments from Kluyveromyces marxianus were cloned in order to identify chromosomal bands in pulsed field electrophoresis patterns of intergeneric hybrid strains which were obtained by protoplast fusion. Molecular hybridization data indicated that the K. marxianus parental strain might be triploid, and it showed strong chromosome length polymorphism. We analyzed the karyotype of two Saccharomyces cerevisiae/K. marxianus hybrid strains (St. 1, St. 46) with our DNA probes and with a Ty1 specific probe. We found indications for recombinational events which lead to the formation of hybrid chromosomal DNA molecules.

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References

  • Bicknell JN, Douglas HC (1970) Nucleic acid homologies among species of Saccharomyces. J Bacteriol 101:505–512

    Google Scholar 

  • Bolivar F, Rodriguez RL, Green PJ, Betlach MC, Heynekker HL, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2:95–113

    Google Scholar 

  • Call HP, Angeneyent J, Emeis CC (1984) PEG- and electrofusion of Kluyveromyces marxianus with Saccharomyces cerevisiae and genetic analysis of fusion products. In: Third European Congress on Biotechnology, vol 3. Verlag Chemie. Weinheim, pp 427–431

    Google Scholar 

  • Carle GF, Olson MV (1984) Separation of chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresis. Nucl Acids Res 12:5647–5664

    Google Scholar 

  • Chan SMC, Tye B-K (1983) Organization of DNA sequences and replication origins at yeast telomeres. Cell 33:563–573

    Google Scholar 

  • Eibel H, Gafner J, Stotz A, Philippsen P (1981) Characterization of the yeast mobile element Ty1. Cold Spring Harbor Symp Quant Biol 45:581–591

    Google Scholar 

  • Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specifity. Anal Biochem 132:6–13

    Google Scholar 

  • Gardiner K, Patterson D (1988) Transverse alternating electrophoresis. Nature 331:371–372

    Google Scholar 

  • Gebeyehu G, Rao PY, Soochan P, Simms DA, Klevan L (1987) Novel biotinylated nucleotide-analogs for labeling and colometric detection of DNA. Nucl Acids Res 15:4513–4534

    Google Scholar 

  • Hoffmann M, Zimmermann M, Emeis CC (1987) Orthogonal field alteration gel electrophoresis (OFAGE) as a means for the analysis of somatic hybrids obtained by protoplast fusion of different Saccharomyces strains. Curr Genet 11:599–603

    Google Scholar 

  • Jonge P de, Jongh FCM de, Mejers R, Steensma HJ, Scheffers WA (1986) Orthogonal-field-alteration gel electrophoresis banding patterns of DNA from yeasts. Yeast 2:193–204

    Google Scholar 

  • Klinner U, Böttcher F (1985) Chromosomal rearrangements after protoplast fusion in the yeast Candida maltosa. Curr Genet 9:619–621

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Ono B, Ishino-Arao Y (1988) Inheritance of chromosome length polymorphisms in Saccharomyces cerevisiae. Curr Genet 14: 413–418

    Google Scholar 

  • Perez C, Vallin C, Benitez J (1984) Hybridization of Saccharomyces cerevisiae with Candida utilis through protoplast fusion. Curr Genet 8:575–580

    Google Scholar 

  • Philippsen P, Eibel H, Gafner J, Stotz A (1983) Ty elements and the stability of the Saccharomyces cerevisiae genome. In: Korhola M, Väisänen E (ed) Proceedings of the ALKO yeast symposium, Found. Biotech. Ind Ferm Res 1, pp 189–200

  • Rigby EWJ, Dieckmann M, Rhodes C, Berg P (1977) Labelling desoxynucleic acid to high specific activity in vitro with nick translation by DNA polymerase I. J Mol Biol 113:237–252

    Google Scholar 

  • Rüther U (1982) pUR250 allows rapid chemical sequencing of both strands of its inserts. Nucl Acid Res 10:5765–5772

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  • Spencer JFT, Spencer DM (1983) Genetic improvement of industrial yeasts. Ann Rev Microbiol 37:121–142

    Google Scholar 

  • Steensma HY, de Jongh FCM, Linnekamp M (1988) The use of electrophoretic karyotypes in the classification of yeasts: Kluyveromyces marxianus and K. lactis. Curr Genet 14:311–317

    Google Scholar 

  • Zimmermann M, Hoffmann-Hintz M, Kolvenbach M, Emeis CC (1988) OFAGE banding patterns of different yeast genera and of intergeneric hybrids. J Basic Microbiol 28:241–247

    Google Scholar 

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Witte, V., Grossmann, B. & Emeis, C.C. Molecular probes for the detection of Kluyveromyces marxianus chromosomal DNA in electrophoretic karyotypes of intergeneric protoplast fusion products. Arch. Microbiol. 152, 441–446 (1989). https://doi.org/10.1007/BF00446926

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  • DOI: https://doi.org/10.1007/BF00446926

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