RRC ID 15303
Author Fujita Y, Giga-Hama Y, Takegawa K.
Title Development of a genetic transformation system using new selectable markers for fission yeast Schizosaccharomyces pombe.
Journal Yeast
Abstract We describe the development of a new transformation system, using multiple auxotrophic marker genes, for the fission yeast Schizosaccharomyces pombe. We developed three new auxotrophic marker genes (arg12(+), tyr1(+) and ade7(+)) and generated a new host strain, YF043, by Cre-loxP-mediated gene disruption. YF043 possessed six mutated biosynthetic genes (leu1-32, ura4-M190T, arg12::loxP, tyr1::loxP, ade7::loxP and his2::loxP). The combination of this host strain and the new selectable markers can be used for gene disruption using the same preexisting transformation systems. In addition, Sz. pombe vectors were constructed, containing selectable marker genes that complement the auxotrophies of YF043. These new vectors are available for gene disruption and heterologous protein expression in strain YF043. The new Sz. pombe host strain will be a useful tool for molecular genetic studies of Sz. pombe where multiple recombinant modifications or multiple mutations are needed.
Volume 22(3)
Pages 193-202
Published 2005-2-1
DOI 10.1002/yea.1201
PMID 15704224
MeSH Blotting, Western Cathepsin A / genetics DNA, Fungal / chemistry DNA, Fungal / genetics Gene Expression Regulation, Fungal Genetic Markers / genetics* Genetic Markers / physiology Genetic Vectors / genetics Green Fluorescent Proteins / genetics Microscopy, Interference Mutagenesis, Insertional / methods Schizosaccharomyces / genetics* Schizosaccharomyces / physiology Schizosaccharomyces / ultrastructure Schizosaccharomyces pombe Proteins / genetics Transformation, Genetic / physiology*
IF 3.143
Times Cited 11
Yeast FYP4216(pAL–SK)