Single-step Marker Switching in Schizosaccharomyces pombe Using a Lithium Acetate Transformation Protocol.

ABSTRACT: The ability to utilize different selectable markers for tagging or mutating multiple genes in Schizosaccharomyces pombe is hampered by the historical use of only two selectable markers, ura4+ and kanMX6; 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 to ura4+ and kanMX6 which 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 fo

SUBMITTER: Brown SD 

PROVIDER: S-EPMC5366253 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature