Skip to main content
Article thumbnail
Location of Repository

A simple and effective chromosome modification method for large-scale deletion of genome sequences and identification of essential genes in fission yeast

By Kyotaro Hirashima, Tomoko Iwaki, Kaoru Takegawa, Yuko Giga-Hama and Hideki Tohda


The technologies for chromosome modification developed to date are not satisfactorily universal, owing to the typical requirements for special enzymes and sequences. In the present report, we propose a new approach for chromosome modification in Schizosaccharomyces pombe that does not involve any special enzymes or sequences. This method, designated the ‘Latour system’, has wide applicability with extremely high efficiency, although both the basic principle and the operation are very simple. We demonstrate the ability of the Latour system to discriminate essential genes, with a long chromosomal area of 100 kb containing 33 genes deleted simultaneously and efficiently. Since no foreign sequences are retained after deletion using the Latour system, this system can be repeatedly applied at other sites. Provided that a negative selectable marker is available, the Latour system relies solely upon homologous recombination, which is highly conserved in living organisms. For this reason, it is expected that the system will be applicable to various yeasts

Topics: Methods Online
Publisher: Oxford University Press
OAI identifier:
Provided by: PubMed Central
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • http://www.pubmedcentral.nih.g... (external link)
  • Suggested articles


    1. (2000). A recyclable Candida albicans URA3 cassette for PCR product-directed gene disruptions.
    2. (2003). Cloning and disruption of the PpURA5 gene and construction of a set of integration vectors for the stable genetic modification of Pichia pastoris.
    3. (2002). DNA mismatch repair and mutation avoidance pathways.
    4. (2005). Efficient and seamless DNA recombineering using a thymidylate synthase A selection system in Escherichia coli.
    5. (1988). Genetic engineeringofSchizosaccharomycespombe:asystemforgenedisruption and replacement using the ura4 gene as a selectable marker.
    6. (1990). High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe.
    7. (2004). Highly efficient gene replacements in Neurospora strains deficient for nonhomologous end-joining.
    8. Langle-Rouault,F.andJacobs,E.(1995)Amethodforperformingprecise alterations in the yeast genome using a recycable selectable marker.
    9. (1999). Markerless gene replacement in Escherichia coli stimulated by a double-strand break in the chromosome.
    10. Nobrega,M.A.,Zhu,Y.,Plajzer-Frick,I.,Afzal,V.andRubin,E.M.(2004) Megabase deletions of gene deserts result in viable mice.
    11. (2000). One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products.
    12. (2002). PCR-mediated generation of a gene disruption construct without the use of DNA ligase and plasmid vectors.
    13. (1999). The URA5 gene encoding orotate-phosphoribosyl transferase of the yeast Kluyveromyces lactis: cloning, sequencing and use as a selectable marker.

    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.