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Applied Microbiology and Biotechnology

, Volume 42, Issue 2–3, pp 353–357 | Cite as

Construction of squalene-accumulatingSaccharomyces cerevisiae mutants by gene disruption through homologous recombination

  • N. Kamimura
  • M. Hidaka
  • H. Masaki
  • T. Uozumi
Original Paper
  • 76 Downloads

Abstract

Saccharomyces cerevisiae synthesizes ergosterol via squalene, but squalene is hardly detected in aerobically grown cells. To obtain a stable squalene-accumulating yeast strain, we attempted to disrupt a gene required in the conversion of squalene to ergosterol, by homologous recombination with a short piece of the gene fragment conjugated with an integration plasmid vector carrying theLEU2 gene. Two mutants that required ergosterol at least for fast growth were isolated. In an aerobic cultivation and with ergosterol supplementation, the two mutants accumulated squalene up to 5 mg/g dry cells. Southern hybridization analysis indicated that both mutants had acquired the vector DNA integrated in the same gene, or nearby genes, on chromosome 12.

Keywords

Saccharomyces Cerevisiae Saccharomyces Homologous Recombination Yeast Strain Gene Fragment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • N. Kamimura
    • 1
  • M. Hidaka
    • 2
  • H. Masaki
    • 2
  • T. Uozumi
    • 2
  1. 1.Petroleum Research LaboratoryMitsubishi Oil Co. Ltd.KanagawaJapan
  2. 2.Department of Biotechnology, Faculty of AgricultureThe University of TokyoTokyoJapan

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