Tools for the genetic manipulation of Zygosaccharomyces rouxii
Language English Country Great Britain, England Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
17887999
DOI
10.1111/j.1567-1364.2007.00308.x
PII: FYR308
Knihovny.cz E-resources
- MeSH
- Escherichia coli genetics MeSH
- Genetic Vectors MeSH
- Mutagenesis, Insertional methods MeSH
- Cloning, Molecular methods MeSH
- Metabolic Networks and Pathways genetics MeSH
- Molecular Biology methods MeSH
- Plasmids MeSH
- Genes, Reporter MeSH
- Sequence Deletion MeSH
- Green Fluorescent Proteins genetics MeSH
- Zygosaccharomyces genetics MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Green Fluorescent Proteins MeSH
A set of tools for the genetic manipulation of the osmotolerant yeast Zygosaccharomyces rouxii was developed. Auxotrophic mutants (ura3 leu2, ura3 ade2, ura3 leu2 ade2) derived from the CBS 732 type strain were prepared. Centromeric and episomal Z. rouxii/Escherichia coli shuttle plasmids with different marker genes (ScURA3, ZrLEU2, ZrADE2) and with multiple cloning sites were constructed, together with a plasmid enabling green fluorescent protein-tagging. A system for repeatable targeted gene deletion in Z. rouxii was established, involving first the integration of a PCR-generated loxP-kanMX-loxP cassette and second the removal of kanMX from the genome using a Z. rouxii plasmid harbouring cre recombinase.
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