Open AccessAn efficient method to optimize Kluyveromyces lactis gene targeting
Author(s) -
WésolowskiLouvel Micheline
Publication year - 2011
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1111/j.1567-1364.2011.00741.x
Subject(s) - biology , kluyveromyces lactis , homologous recombination , plasmid , genetics , gene targeting , cre recombinase , recombinase , gene , mutant , locus (genetics) , cre lox recombination , homologous chromosome , dna , kluyveromyces , genome , crispr , computational biology , saccharomyces cerevisiae , recombination , transgene , genetically modified mouse
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 lox P‐ kan MX‐ lox P cassette together with a Cre ‐recombinase plasmid, a nej1 ∷ lox P 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.