A simple and effective chromosome modification method for large-scale deletion of genome sequences and identification of essential genes in fission yeast.
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.
Hirashima
Kyotaro
K
ASPEX Division, Asahi Glass Co., Ltd Hazawa-cho, Yokohama, Kanagawa 221-8755, Japan.
Iwaki
Tomoko
T
Takegawa
Kaoru
K
Giga-Hama
Yuko
Y
Tohda
Hideki
H
D050539:3-Isopropylmalate Dehydrogenase / Q000235:genetics; D015825:Chromosomes, Fungal; D017353:Gene Deletion; D005800:Genes, Fungal; D005819:Genetic Markers; D023281:Genomics / Q000379:methods*; D009154:Mutation; D012568:Schizosaccharomyces / Q000235:genetics* / Q000254:growth & development; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics
D005819:Genetic Markers; D029702:Schizosaccharomyces pombe Proteins; C400810:ura4 protein, S pombe; D050539:3-Isopropylmalate Dehydrogenase