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dc.contributor.authorMacIver, Fiona H
dc.contributor.authorGlover, David M
dc.contributor.authorHagan, Iain M
dc.date.accessioned2009-09-23T09:23:03Z
dc.date.available2009-09-23T09:23:03Z
dc.date.issued2003-05
dc.identifier.citationA 'marker switch' approach for targeted mutagenesis of genes in Schizosaccharomyces pombe. 2003, 20 (7):587-94 Yeasten
dc.identifier.issn0749-503X
dc.identifier.pmid12734797
dc.identifier.doi10.1002/yea.983
dc.identifier.urihttp://hdl.handle.net/10541/82220
dc.description.abstractThe completion of the Schizosaccharomyces pombe genome sequencing project has led to a dramatic acceleration of gene characterization in this system. Once a gene has been identified, the challenge then comes in using reverse genetics to generate a range of mutants in this gene of interest so that the powerful genetics and wealth of genetic backgrounds available in Sz. pombe can be exploited to study the function of the newly identified molecule. Beyond simple PCR-tagging approaches, the high frequency with which illegitimate recombination occurs in Sz. pombe has made the manipulation of some loci complex, time consuming and a process of trial and error. Here we describe a simple 'marker switch' approach that enables the rapid selection of integration events at the locus of interest from an excessive background of integration at heterologous sites. We use the generation of temperature-sensitive mutations in the plo1(+) gene to validate this approach.
dc.language.isoenen
dc.subject.meshBase Sequence
dc.subject.meshDNA, Fungal
dc.subject.meshDrosophila Proteins
dc.subject.meshGenes, Fungal
dc.subject.meshGenetic Markers
dc.subject.meshMutagenesis, Site-Directed
dc.subject.meshPhenotype
dc.subject.meshPlasmids
dc.subject.meshProtein-Serine-Threonine Kinases
dc.subject.meshSchizosaccharomyces
dc.subject.meshTemperature
dc.titleA 'marker switch' approach for targeted mutagenesis of genes in Schizosaccharomyces pombe.en
dc.typeArticleen
dc.contributor.departmentPaterson Institute for Cancer Research, Wilmslow Road, Manchester M20 4BX, UK.en
dc.identifier.journalYeasten
html.description.abstractThe completion of the Schizosaccharomyces pombe genome sequencing project has led to a dramatic acceleration of gene characterization in this system. Once a gene has been identified, the challenge then comes in using reverse genetics to generate a range of mutants in this gene of interest so that the powerful genetics and wealth of genetic backgrounds available in Sz. pombe can be exploited to study the function of the newly identified molecule. Beyond simple PCR-tagging approaches, the high frequency with which illegitimate recombination occurs in Sz. pombe has made the manipulation of some loci complex, time consuming and a process of trial and error. Here we describe a simple 'marker switch' approach that enables the rapid selection of integration events at the locus of interest from an excessive background of integration at heterologous sites. We use the generation of temperature-sensitive mutations in the plo1(+) gene to validate this approach.


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