A 'marker switch' approach for targeted mutagenesis of genes in Schizosaccharomyces pombe.

2.50
Hdl Handle:
http://hdl.handle.net/10541/82220
Title:
A 'marker switch' approach for targeted mutagenesis of genes in Schizosaccharomyces pombe.
Authors:
MacIver, Fiona H; Glover, David M; Hagan, Iain M
Abstract:
The 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.
Affiliation:
Paterson Institute for Cancer Research, Wilmslow Road, Manchester M20 4BX, UK.
Citation:
A 'marker switch' approach for targeted mutagenesis of genes in Schizosaccharomyces pombe. 2003, 20 (7):587-94 Yeast
Journal:
Yeast
Issue Date:
May-2003
URI:
http://hdl.handle.net/10541/82220
DOI:
10.1002/yea.983
PubMed ID:
12734797
Type:
Article
Language:
en
ISSN:
0749-503X
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
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.en
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
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