Applied Microbiology and Biotechnology

, Volume 72, Issue 5, pp 947–952

Chromosome-shuffling technique for selected chromosomal segments in Saccharomyces cerevisiae

  • Minetaka Sugiyama
  • Eishi Yamamoto
  • Yukio Mukai
  • Yoshinobu Kaneko
  • Masafumi Nishizawa
  • Satoshi Harashima
Applied Genetics and Molecular Biotechnology

Abstract

We describe a novel chromosome engineering technique for shuffling selected regions of chromosomes from two strains in Saccharomyces cerevisiae: The technique starts with the construction of MATa and MATα strains in which a particular chromosome is split at exactly the same site in both strains such that the split chromosomes generated are marked with different markers. The two strains are then crossed, and the resultant diploid is cultivated in nutrient medium to induce loss of the split chromosome originating from either of the strains. We predicted that some of these clones that are hemizygous for the split chromosome would spontaneously restore a homozygous configuration of the split chromosome during cultivation. We verified this prediction by tetrad analysis and quantitative Southern hybridization analysis, indicating that it is possible to create diploid hybrids in which a selected region of a chromosome from one strain is replaced by the corresponding chromosomal region from another strain. We also found that some chromosomal segments maintain a hemizygous state. This novel technique, which we call ‘chromosome shuffling’, could provide a new tool to analyze phenotypic alterations caused by the replacement or hemizygosity of a selected chromosomal region in not only laboratory but also industrial strains of S. cerevisiae.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Minetaka Sugiyama
    • 1
  • Eishi Yamamoto
    • 1
  • Yukio Mukai
    • 1
  • Yoshinobu Kaneko
    • 1
  • Masafumi Nishizawa
    • 2
  • Satoshi Harashima
    • 1
  1. 1.Department of Biotechnology, Graduate School of EngineeringOsaka UniversityOsakaJapan
  2. 2.Department of Microbiology and ImmunologyKeio University School of MedicineTokyoJapan

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