Natesuntorn Waranya, Iwami Kotaro, Matsubara Yuki, Sasano Yu, Sugiyama Minetaka, Kaneko Yoshinobu, Harashima Satoshi
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Sci Rep. 2015 Jul 30;5:12510. doi: 10.1038/srep12510.
Segmental aneuploidy can play an important role in environmental adaptation. However, study of segmental aneuploids is severely hampered by the difficulty of creating them in a designed fashion. Here, we describe a PCR-mediated chromosome duplication (PCDup) technology that enables the generation of segmental aneuploidy at any desired chromosomal region in Saccharomyces cerevisiae. We constructed multiple strains harboring 100 kb to 200 kb segmental duplications covering the whole of the S. cerevisiae genome. Interestingly, some segmental aneuploidies confer stress tolerance, such as to high temperature, ethanol and strong acids, while others induce cell lethality and stress sensitivity, presumably as result of the simultaneous increases in dosages of multiple genes. We suggest that our PCDup technology will accelerate studies into the phenotypic changes resulting from alteration of gene dosage balance of multiple genes and will provide new insights into the adaptive molecular mechanisms in the genome in segmental aneuploidy-derived human diseases.
染色体片段非整倍体在环境适应中可能发挥重要作用。然而,由于难以以设计的方式创建染色体片段非整倍体,对其研究受到严重阻碍。在此,我们描述了一种PCR介导的染色体复制(PCDup)技术,该技术能够在酿酒酵母的任何所需染色体区域产生染色体片段非整倍体。我们构建了多个携带100 kb至200 kb片段重复的菌株,这些重复覆盖了整个酿酒酵母基因组。有趣的是,一些染色体片段非整倍体赋予了对高温、乙醇和强酸等胁迫的耐受性,而其他一些则导致细胞致死和胁迫敏感性,这可能是多个基因剂量同时增加的结果。我们认为,我们的PCDup技术将加速对多个基因剂量平衡改变导致的表型变化的研究,并将为染色体片段非整倍体衍生的人类疾病中基因组的适应性分子机制提供新的见解。