一种可在芽殖酵母中重复使用的新型高效基因破坏盒。
A new efficient gene disruption cassette for repeated use in budding yeast.
作者信息
Güldener U, Heck S, Fielder T, Beinhauer J, Hegemann J H
机构信息
Institut für Mikrobiologie und Molekularbiologie, Justus-Liebig-Universität Giessen, Giessen, Germany.
出版信息
Nucleic Acids Res. 1996 Jul 1;24(13):2519-24. doi: 10.1093/nar/24.13.2519.
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.
摘要
显性卡那霉素抗性（kanr）标记基因在芽殖酵母的基因破坏实验中起着重要作用，因为该标记可用于多种缺乏传统酵母标记的酵母菌株。我们开发了一种loxP-kanMX-loxP基因破坏盒，它结合了异源卡那霉素抗性标记与Cre-loxP重组系统的优点。这个破坏盒通过同源整合高效地整合到正确的基因组位点（通常为70%）。当Cre重组酶表达时，kanMX模块通过loxP位点之间的有效重组被切除，在染色体位点上留下一个单一的loxP位点。该系统允许卡那霉素抗性标记基因的重复使用，这对于基因家族的功能分析将具有很大优势。
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