Wei Min, Madia Federica, Longo Valter D
Andrus Gerontology Center, Department of Biological Sciences, University of Southern California, Los Angeles, USA.
J Vis Exp. 2011 Sep 29(55):3030. doi: 10.3791/3030.
Studies using the Saccharomyces cerevisiae aging model have uncovered life span regulatory pathways that are partially conserved in higher eukaryotes. The simplicity and power of the yeast aging model can also be explored to study DNA damage and genome maintenance as well as their contributions to diseases during aging. Here, we describe a system to study age-dependent DNA mutations, including base substitutions, frame-shift mutations, gross chromosomal rearrangements, and homologous/homeologous recombination, as well as nuclear DNA repair activity by combining the yeast chronological life span with simple DNA damage and mutation assays. The methods described here should facilitate the identification of genes/pathways that regulate genomic instability and the mechanisms that underlie age-dependent DNA mutations and cancer in mammals.
利用酿酒酵母衰老模型开展的研究揭示了在高等真核生物中部分保守的寿命调控途径。酵母衰老模型的简易性和实用性还可用于研究DNA损伤与基因组维持，以及它们在衰老过程中对疾病的影响。在此，我们描述了一个用于研究年龄依赖性DNA突变的系统，包括碱基替换、移码突变、染色体大片段重排以及同源/异源重组，还可通过将酵母时序寿命与简单的DNA损伤和突变检测相结合来研究核DNA修复活性。本文所述方法应有助于鉴定调控基因组不稳定的基因/途径，以及哺乳动物中年龄依赖性DNA突变和癌症背后的机制。
