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

Authors
  • Foureau, Emilien1
  • Courdavault, Vincent2
  • Rojas, Luisa Fernanda3
  • Dutilleul, Christelle2
  • Simkin, Andrew J.4
  • Crèche, Joël1
  • Atehortùa, Lucia3
  • Giglioli-Guivarc’h, Nathalie2
  • Clastre, Marc1
  • Papon, Nicolas1
  • 1 Université François-Rabelais de Tours, EA2106, Biomolécules et Biotechnologies Végétales, Faculté de Pharmacie, Tours, France , Tours (France)
  • 2 Université François-Rabelais de Tours, EA2106, Biomolécules et Biotechnologies Végétales, Faculté des Sciences et Techniques, Tours, France , Tours (France)
  • 3 Universidad de Antioquia, Laboratorio de Biotecnología, Sede de Investigación Universitaria, Medellín, Colombia , Medellín (Colombia)
  • 4 University of Essex, School of Biological Sciences, Wivenhoe Park, Colchester, UK , Colchester (United Kingdom)
Type
Published Article
Journal
Biotechnology Letters
Publisher
Springer-Verlag
Publication Date
Mar 05, 2013
Volume
35
Issue
7
Pages
1035–1043
Identifiers
DOI: 10.1007/s10529-013-1169-7
Source
Springer Nature
Keywords
License
Yellow

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. guilliermondiiku70 deficient mutant thus represents a powerful recipient strain to knock-out genes efficiently in this yeast.

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