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Nucleic Acids Research 2006 34(2):e11; doi:10.1093/nar/gnj011
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Published online 24 January 2006

© The Author 2006. Published by Oxford University Press. All rights reserved
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Methods Online

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

Kyotaro Hirashima, Tomoko Iwaki1, Kaoru Takegawa1, Yuko Giga-Hama and Hideki Tohda*

ASPEX Division, Asahi Glass Co., Ltd Hazawa-cho, Yokohama, Kanagawa 221-8755, Japan 1Department of Life Sciences, Faculty of Agriculture, Kagawa University Miki-cho, Kagawa 761-0795, Japan

*To whom correspondence should be addressed. Tel: +81 45 374 7377; Fax: +81 45 374 8872; Email: htohda{at}anet.ne.jp

Received December 2, 2005. Revised December 27, 2005. Accepted January 4, 2006.

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.


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