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Efficient gene targeting in a Candida guilliermondii non-homologous end-joining pathway-deficient strain

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Abstract

The yeast, Candida guilliermondii, has been widely studied due to its biotechnological interest as well as its biological control potential. It integrates foreign DNA predominantly via ectopic events, likely through the well-known non-homologous end-joining (NHEJ) pathway involving the Ku70p/Ku80p heterodimer, Lig4p, Nej1p and Lif1p. This phenomenon remains highly deleterious for targeted gene knock-out strategies that require the homologous recombination process. Here, we have constructed a ku70 mutant strain derived from the ATCC 6260 reference strain of C. guilliermondii. Following a series of disruption attempts of various genes (FCY1, ADE2 and TRP5), using several previously described dominant selectable markers (URA5, SAT-1 and HPH #), we demonstrated that the efficiencies of homologous gene targeting in such a NHEJ-deficient strain was very high compared to the wild type strain. The C. guilliermondii ku70 deficient mutant thus represents a powerful recipient strain to knock-out genes efficiently in this yeast.

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Acknowledgments

We acknowledge the Broad Institute Fungal Genome Initiative for making the complete genome sequence of C. guilliermondii available. We thank Stéphane Marcand (Commissariat à l’Energie Atomique, Laboratoire Télomère et Réparation du Chromosome, Fontenay-aux-Roses, France) for precious advices.

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Authors and Affiliations

  1. EA2106, Biomolécules et Biotechnologies Végétales, Faculté de Pharmacie, Université François-Rabelais de Tours, Tours, France

    Emilien Foureau, Joël Crèche, Marc Clastre & Nicolas Papon

  2. EA2106, Biomolécules et Biotechnologies Végétales, Faculté des Sciences et Techniques, Université François-Rabelais de Tours, Tours, France

    Vincent Courdavault, Christelle Dutilleul & Nathalie Giglioli-Guivarc’h

  3. Laboratorio de Biotecnología, Sede de Investigación Universitaria, Universidad de Antioquia, Medellín, Colombia

    Luisa Fernanda Rojas & Lucia Atehortùa

  4. School of Biological Sciences, University of Essex, Wivenhoe Park, Colchester, UK

    Andrew J. Simkin

Authors
  1. Emilien Foureau
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  2. Vincent Courdavault
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  3. Luisa Fernanda Rojas
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  4. Christelle Dutilleul
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  5. Andrew J. Simkin
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  6. Joël Crèche
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  7. Lucia Atehortùa
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  8. Nathalie Giglioli-Guivarc’h
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  9. Marc Clastre
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  10. Nicolas Papon
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Corresponding author

Correspondence to Nicolas Papon.

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Foureau, E., Courdavault, V., Rojas, L.F. et al. Efficient gene targeting in a Candida guilliermondii non-homologous end-joining pathway-deficient strain. Biotechnol Lett 35, 1035–1043 (2013). https://doi.org/10.1007/s10529-013-1169-7

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  • DOI: https://doi.org/10.1007/s10529-013-1169-7

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