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 Iwaki, Kaoru Takegawa, Yuko Giga-Hama, Hideki Tohda

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
JournalNucleic Acids Research
Volume34
Issue number2
Publication statusPublished - 2006
Externally publishedYes

Fingerprint

Schizosaccharomyces
Sequence Deletion
Essential Genes
Chromosomes
Genome
Homologous Recombination
Enzymes
Yeasts
Technology
Genes

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

A simple and effective chromosome modification method for large-scale deletion of genome sequences and identification of essential genes in fission yeast. / Hirashima, Kyotaro; Iwaki, Tomoko; Takegawa, Kaoru; Giga-Hama, Yuko; Tohda, Hideki.

In: Nucleic Acids Research, Vol. 34, No. 2, 2006.

Research output: Contribution to journalArticle

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