Cassette for the generation of sequential gene disruptions in the yeast Schizosaccharomyces pombe

BioTechniques
D S McNabbL Guarente

Abstract

The ability to conveniently construct gene disruptions is an important methodology for genetic analysis of the fission yeast Schizosaccharomyces pombe. Because of the limited number of selectable markers available for generating gene disruptions in fission yeast, the construction of strains that contain multiple gene disruptions can be quite difficult. This becomes a particular problem when episomal plasmids carrying selectable markers are also required within the same strains. To alleviate these difficulties, we have constructed a hisG-ura(4+)-hisG cassette that can be used repeatedly for constructing gene disruptions in S. pombe. This cassette allows the recycling of the ura4+ marker, thereby permitting the disruption of an indefinite number of genes sequentially within the same strain and/or for subsequently introducing a ura(4+)-marked plasmid.

Citations

May 21, 2009·Applied Microbiology and Biotechnology·Shohei KanekoAkihiko Kondo
Nov 9, 2016·BMC Biotechnology·Hanna AmelinaKazunori Tomita
Aug 14, 1999·Nucleic Acids Research·M LombaertsJ Brouwer
Feb 10, 2006·Eukaryotic Cell·Jude Beaudoin, Simon Labbé
Sep 24, 2002·The Journal of Biological Chemistry·Daniel R BellemareSimon Labbe
Feb 1, 2000·Biochemical and Biophysical Research Communications·M LombaertsJ Brouwer
Dec 6, 2000·Mutation Research· van den Bosch MA Pastink
Nov 22, 2005·Biochimica Et Biophysica Acta·Csaba FenyvuesvolgyiRichard Yuqi Zhao

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