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A new efficient gene disruption cassette for repeated use in budding yeast
U Güldener1, S Heck, T Fielder
1Institut für Mikrobiologie und Molekularbiologie, Justus-Liebig-Universität Giessen, Giessen, Germany.
Nucleic Acids Research
|July 1, 1996
Summary
This study introduces a new gene disruption cassette for budding yeast, combining a kanamycin resistance (kanr) marker with the Cre-lox P system. This system allows for efficient gene disruption and repeated marker use, aiding in the analysis of gene families.
Area of Science:
- Molecular Biology
- Yeast Genetics
Background:
- The kanamycin resistance (kanr) gene is a valuable marker in budding yeast gene disruption.
- Conventional markers are not always suitable for all yeast strains.
Purpose of the Study:
- To develop a novel gene disruption cassette for budding yeast.
- To combine the benefits of the kanr marker with the Cre-lox P recombination system.
Main Methods:
- Development of a loxP-kanMX-loxP gene disruption cassette.
- Utilizing homologous integration for cassette insertion into the yeast genome.
- Employing Cre recombinase for excision of the kanMX module.
Main Results:
- The disruption cassette integrates with high efficiency (routinely 70%) at the correct genomic locus.
- Cre recombinase efficiently excises the kanMX module, leaving a single loxP site.
- The system enables repeated use of the kanr marker gene.
Conclusions:
- The loxP-kanMX-loxP cassette offers an efficient method for gene disruption in yeast.
- This system facilitates the functional analysis of gene families through repeated marker application.