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. 1996 Jul 1;24(13):2519-24.
doi: 10.1093/nar/24.13.2519.

A new efficient gene disruption cassette for repeated use in budding yeast

U Güldener  1 , S Heck, T Fielder, J Beinhauer, J H Hegemann
Affiliations

A new efficient gene disruption cassette for repeated use in budding yeast

U Güldener et al. Nucleic Acids Res. .

Abstract

The dominant kanr marker gene plays an important role in gene disruption experiments in budding yeast, as this marker can be used in a variety of yeast strains lacking the conventional yeast markers. We have developed a loxP-kanMX-loxP gene disruption cassette, which combines the advantages of the heterologous kanr marker with those from the Cre-lox P recombination system. This disruption cassette integrates with high efficiency via homologous integration at the correct genomic locus (routinely 70%). Upon expression of the Cre recombinase the kanMX module is excised by an efficient recombination between the loxP sites, leaving behind a single loxP site at the chromosomal locus. This system allows repeated use of the kanr marker gene and will be of great advantage for the functional analysis of gene families.

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