Development of a genetic transformation system using new selectable markers for fission yeast Schizosaccharomyces pombe

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...

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Bibliographic Details
Published inYeast (Chichester, England) Vol. 22; no. 3; pp. 193 - 202
Main Authors Fujita, Yasuko, Giga‐Hama, Yuko, Takegawa, Kaoru
Format Journal Article
LanguageEnglish
Published Chichester, UK John Wiley & Sons, Ltd 01.02.2005
Subjects
auxotrophic marker
Blotting, Western
Cathepsin A - genetics
Cre‐loxP
DNA, Fungal - chemistry
DNA, Fungal - genetics
fission yeast
Gene Expression Regulation, Fungal
Genetic Markers - genetics
Genetic Markers - physiology
Genetic Vectors - genetics
Green Fluorescent Proteins - genetics
heterologous expression
Microscopy, Interference
Mutagenesis, Insertional - methods
Schizosaccharomyces - genetics
Schizosaccharomyces - physiology
Schizosaccharomyces - ultrastructure
Schizosaccharomyces pombe
Schizosaccharomyces pombe Proteins - genetics
transformation system
Transformation, Genetic - physiology
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Summary: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. Copyright © 2005 John Wiley & Sons, Ltd.
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
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ISSN:0749-503X
1097-0061
DOI:10.1002/yea.1201