Open AccessTools for the genetic manipulation of Zygosaccharomyces rouxii
Author(s) -
Pribylova Lenka,
De Montigny Jacky,
Sychrova Hana
Publication year - 2007
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.1567-1364.2007.00308.x
Subject(s) - ura3 , plasmid , biology , escherichia coli , auxotrophy , shuttle vector , multiple cloning site , mutant , genetics , recombinase , gene , microbiology and biotechnology , recombinant dna , recombination , expression vector , vector (molecular biology)
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.