title: Efficient gene targeting in a Candida guilliermondii non-homologous end-joining pathway-deficient strain.
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.
authors: EA2106, Biomolécules et Biotechnologies Végétales, Faculté de Pharmacie, Université François-Rabelais de Tours, Tours, France.
authors: Emilien
authors: E
authors: Foureau
authors: Vincent
authors: V
authors: Courdavault
authors: Luisa Fernanda
authors: LF
authors: Rojas
authors: Christelle
authors: C
authors: Dutilleul
authors: Andrew J
authors: AJ
authors: Simkin
authors: Joël
authors: J
authors: Crèche
authors: Lucia
authors: L
authors: Atehortùa
authors: Nathalie
authors: N
authors: Giglioli-Guivarc'h
authors: Marc
authors: M
authors: Clastre
authors: Nicolas
authors: N
authors: Papon
mesh_terms: D034961:Antigens, Nuclear / Q000235:genetics; D002175:Candida / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D055786:Gene Knockout Techniques; D018390:Gene Targeting / Q000379:methods*; D005827:Genetics, Microbial / Q000379:methods*; D000072200:Ku Autoantigen; D011995:Recombination, Genetic
chemical_list: D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D000072200:Ku Autoantigen