Watanabe Takahito, Murata Yoshinori, Oka Syuichi, Iwahashi Hitoshi
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.
Yeast. 2004 Mar;21(4):351-65. doi: 10.1002/yea.1103.
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.
DNA-DNA杂交被认为是阐明密切相关分类群(如物种和菌株)之间关系的 superior 方法。对于物种鉴定,我们提出了一种新的DNA-DNA杂交方法:基于DNA微阵列的比较基因组杂交(CGH)方法,使用含有约6000个基因的酵母DNA微阵列。将来自酵母菌株作为样本菌株(样本)的基因组用Cy3染料标记,并与作为参考菌株(参考)的酿酒酵母S288C的Cy5标记基因组一起与单个DNA微阵列杂交。估计所有基因点信号强度的log2比值[log2[Cy3(样本)/Cy5(参考)]:比值],并分为以下几组:比值≤-1;-1<比值<1;1≤比值。属于酿酒酵母属的模式菌株基因组的杂交图谱与酿酒酵母S288C的杂交图谱有显著差异。基于比值的分组使我们能够更清楚地区分一些与酿酒酵母不同的物种。此外,聚类分析区分了密切相关的物种和菌株。使用这种方法,我们不仅能够进行物种鉴定,还能够以单基因分辨率获得有关此类基因扩增和缺失的基因拷贝数变化的信息。这些观察结果表明,基于DNA微阵列的CGH是一种用于物种鉴定和比较基因组分析的强大系统。