A new approach to species determination for yeast strains: DNA microarray-based comparative genomic hybridization using a yeast DNA microarray with 6000 genes.
DNA-DNA hybridization is known as the superior method in the elucidation of relationships between closely related taxa, such as species and strain. For species determination we propose a new DNA-DNA hybridization method: the DNA microarray-based comparative genomic hybridization (CGH) method, using a yeast DNA microarray with approximately 6000 genes. The genome from a yeast strain as a sample strain (Sample) was labelled with Cy3-dye and hybridized to a single DNA microarray, together with the Cy5-labelled genome of S. cerevisiae S288C as a reference strain (Reference). The log2 ratio values [log2[Cy3(Sample)/Cy5(Reference)]: Ratio] of signal intensities of all the gene spots were estimated and divided into the following groups: Ratio < or = -1; -1 < Ratio < 1; 1 < or = Ratio. The hybridization profiles of the genomes of type strains belonging to the genus Saccharomyces were significantly different from that of S. cerevisiae S288C. The Ratio-based grouping allowed us to discriminate between some species from S. cerevisiae more clearly. Furthermore, cluster analysis discriminated between closely related species and strains. Using this method, we were able to not only perform species determination but also to obtain information on alternation in gene copy number of such gene amplifications and deletions with single-gene resolution. These observations indicated that DNA microarray-based CGH is a powerful system for species determination and comparative genome analysis.
Watanabe
Takahito
T
International Patent Organism Depositary, National Institute of Advanced Industrial Science and Technology (AIST), AIST Central 6, 1-1-1, Higashi, Tsukuba, Ibaraki 305-8566, Japan. takahito@kuwri.kyoto-u.ac.jp
Murata
Yoshinori
Y
Oka
Syuichi
S
Iwahashi
Hitoshi
H
D016000:Cluster Analysis; D004271:DNA, Fungal / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D016681:Genome, Fungal; D009693:Nucleic Acid Hybridization / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010802:Phylogeny; D012331:RNA, Fungal / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000145:classification* / Q000235:genetics*; D015003:Yeasts / Q000145:classification* / Q000235:genetics*
D004271:DNA, Fungal; D004275:DNA, Ribosomal; D012331:RNA, Fungal; D012336:RNA, Ribosomal, 16S