title: An efficient method to optimize Kluyveromyces lactis gene targeting.
abstract: Kluyveromyces lactis strains impaired in the nonhomologous end-joining pathway are relevant tools for the homologous integration of exogenous DNA into the genome, as in the mutant strains, close to 100% of the integrants are targeted to the homologous locus, compared with a few per cent for the wild-type recipient. Using a loxP-kanMX-loxP cassette together with a Cre-recombinase plasmid, a nej1∷loxP mutant strain suitable for multiple gene disruption has been constructed. Furthermore, using this strain, PCR-generated constructs with only 50 bp of homologous flanking sequences resulted in efficient exogenous DNA targeting.
authors.affiliation: Génétique Moléculaire des Levures, UMR Microbiologie Adaptation et Pathogénie, Université de Lyon, France. micheline.louvel@univ-lyon1.fr
authors.forename: Micheline
authors.initials: M
authors.lastname: Wésolowski-Louvel
mesh_terms: D018390:Gene Targeting / Q000379:methods*; D005827:Genetics, Microbial / Q000379:methods*; D007716:Kluyveromyces / Q000235:genetics*; D016254:Mutagenesis, Insertional / Q000379:methods; D011995:Recombination, Genetic