title: Single-step Marker Switching inSchizosaccharomyces pombeUsing a Lithium Acetate Transformation Protocol.
abstract: The ability to utilize different selectable markers for tagging or mutating multiple genes inSchizosaccharomyces pombeis hampered by the historical use of only two selectable markers,ura4+andkanMX6; the latter conferring resistance to the antibiotic G418 (geneticin). More markers have been described recently, but introducing these into yeast cells often requires strain construction from scratch. To overcome this problem we and other groups have created transformation cassettes with flanking homologies toura4+andkanMX6which enable an efficient and time-saving way to exchange markers in existing mutated or tagged fission yeast strains. Here, we present a protocol for single-step marker switching by lithium acetate transformation in fission yeast,Schizosaccharomyces pombe. In the following we describe how to swap theura4+marker to akanMX6, natMX4,orhphMX4marker, which provide resistance against the antibiotics G418, nourseothricin (clonNAT) or hygromycin B, respectively. We also detail how to exchange any of theMXmarkers for nutritional markers, such asarg3+, his3+, leu1+andura4+.
authors.affiliation: Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
authors.forename: Simon D
authors.initials: SD
authors.lastname: Brown
authors.affiliation: Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
authors.forename: Alexander
authors.initials: A
authors.lastname: Lorenz
keywords: Gene deletion; Gene tagging; Genetic manipulation; Li-Acetate transformation; Marker switch; Schizosaccharomyces pombe; Selectable marker