Department of Genetics, Biology of Microorganisms, Anthropology, Evolution, Viale Usberti 11/A, 43124 Parma, Italy.
Methods. 2010 Aug;51(4):426-36. doi: 10.1016/j.ymeth.2010.02.023. Epub 2010 Mar 2.
The yeast Saccharomyces cerevisiae has the capacity to survive large deletions or total loss of mtDNA (petite mutants), and thus in the last few years it has been used as a model system to study defects in mitochondrial DNA (mtDNA) maintenance produced by mutations in genes involved in mtDNA replication. In this paper we describe methods to obtain strains harboring mutations in nuclear genes essential for the integrity of mtDNA, to measure the frequency and the nature of petite mutants, to estimate the point mutation frequency in mtDNA and to determine whether a nuclear mutation is recessive or dominant and, in the latter case, the kind of dominance.
酿酒酵母(Saccharomyces cerevisiae)具有在 mtDNA( petite 突变体)大量缺失或完全缺失的情况下存活的能力,因此,在过去几年中,它一直被用作研究涉及 mtDNA 复制基因发生突变引起的 mtDNA 维持缺陷的模型系统。在本文中,我们描述了获得含有核基因突变体的菌株的方法,这些核基因突变体对于 mtDNA 的完整性是必需的,用于测量 petite 突变体的频率和性质,估计 mtDNA 中点突变频率,并确定核基因突变是隐性还是显性,如果是后者,则确定显性的类型。