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

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    Authors
    MacIver, Fiona H
    Glover, David M
    Hagan, Iain M
    Affiliation
    Paterson Institute for Cancer Research, Wilmslow Road, Manchester M20 4BX, UK.
    Issue Date
    2003-05
    
    Metadata
    Show full item record
    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.
    Citation
    A 'marker switch' approach for targeted mutagenesis of genes in Schizosaccharomyces pombe. 2003, 20 (7):587-94 Yeast
    Journal
    Yeast
    URI
    http://hdl.handle.net/10541/82220
    DOI
    10.1002/yea.983
    PubMed ID
    12734797
    Type
    Article
    Language
    en
    ISSN
    0749-503X
    ae974a485f413a2113503eed53cd6c53
    10.1002/yea.983
    Scopus Count
    Collections
    All Paterson Institute for Cancer Research

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