Increased homologous integration frequency in Yarrowia lipolytica strains defective in non-homologous end-joining View Full Text


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Article Info

DATE

2013-02-20

AUTHORS

Anne Kretzschmar, Christina Otto, Martina Holz, Severine Werner, Linda Hübner, Gerold Barth

ABSTRACT

The ascomycetous yeast Yarrowia lipolytica has been established as model system for studies of several research topics as well as for biotechnological processes in the last two decades. However, frequency of heterologous recombination is high in this yeast species, and so knockouts of genes are laborious to achieve. Therefore, the aim of this study was to check whether a reduction of non-homologous end-joining (NHEJ) of double strand breaks (DSB) results in a strong increase of proportion of homologous recombinants. The Ku70–Ku80 heterodimer is known as an essential protein complex of the NHEJ. We show that deletion of YlKU70 and/or YlKU80 results in an increase of the rate of transformants with homologous recombination (HR) up to 85 % in each case. However, it never reaches near 100 % of HR in any case as described for some other yeast. Furthermore, we demonstrated that growth of Δylku strains was similar to that of the wild-type strain. In addition, no differences were detected between the Δylku strains and the parent strain in respect to sensitivity to the mutagenic agent EMS as well as to the antibiotics hygromycin, bleomycin and nourseothricin. However, Δylku70 and Δylku80 strain showed a slightly higher sensitivity against UV rays. Thus, the new constructed Δylku strains are attractive recipient strains for homologous integration of DNA fragments and a valuable tool for directed knockouts of genes. Nevertheless, our data suggest the existence of another system of non-homologous recombination what may be subject of further investigation. More... »

PAGES

63-72

References to SciGraph publications

  • 2012-12-09. Efficient homologous recombination with short length flanking fragments in Ku70 deficient Yarrowia lipolytica strains in BIOTECHNOLOGY LETTERS
  • 2003-11-01. The Neurospora crassa mus-19 gene is identical to the qde-3 gene, which encodes a RecQ homologue and is involved in recombination repair and postreplication repair in CURRENT GENETICS
  • 2008-10-11. Aconitase overexpression changes the product ratio of citric acid production by Yarrowia lipolytica in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2007-04-20. Citric acid production from sucrose using a recombinant strain of the yeast Yarrowia lipolytica in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2004-07. Genome evolution in yeasts in NATURE
  • 2003-07-10. Functional genetics of Yarrowia lipolytica in FUNCTIONAL GENETICS OF INDUSTRIAL YEASTS
  • 1996. Yarrowia lipolytica in NONCONVENTIONAL YEASTS IN BIOTECHNOLOGY
  • 2010-10-30. Overexpression of alpha-ketoglutarate dehydrogenase in Yarrowia lipolytica and its effect on production of organic acids in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2011-10-02. Overproduction and secretion of α-ketoglutaric acid by microorganisms in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2007-10-19. Overexpression of the ICL1 gene changes the product ratio of citric acid production by Yarrowia lipolytica in APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
  • 2012-02-18. The Yeast Yarrowia lipolytica as a Generic Tool for Molecular Evolution of Enzymes in LIPASES AND PHOSPHOLIPASES
  • 2003. Induction of Citric Acid/lsocitric Acid and α-Ketoglutaric Acid Production in the Yeast Yarrowia lipolytica in NON-CONVENTIONAL YEASTS IN GENETICS, BIOCHEMISTRY AND BIOTECHNOLOGY
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s00294-013-0389-7

    DOI

    http://dx.doi.org/10.1007/s00294-013-0389-7

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1049065515

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/23423527


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