Chiron Stéphane, Gaisne Mauricette, Guillou Emmanuelle, Belenguer Pascale, Clark-Walker G Desmond, Bonnefoy Nathalie
Centre de Génétique Moléculaire, UPR 2167, Gif-sur-Yvette, France.
Methods Mol Biol. 2007;372:91-105. doi: 10.1007/978-1-59745-365-3_7.
The fission yeast Schizosaccharomyces pombe, widely used for studies of cell cycle control and differentiation, provides an alternative and complementary model to the budding yeast Saccharomyces cerevisiae for studies of nucleo-mitochondrial interactions. There are striking similarities between S. pombe and mammalian cells, in both their respiratory physiology and their mitochondrial genome structure. This technical review briefly lists the general and specific properties that are helpful to know when starting to use fission yeast as a model system for mitochondrial studies. In addition, advice is given for cell growth and genetic techniques, tips for disruption of genes involved in respiration are presented. and a basic differential centrifugation protocol is provided for the isolation of purified mitochondria that are suitable for diverse applications such as subfractionation and in vitro import.
裂殖酵母粟酒裂殖酵母被广泛用于细胞周期调控和分化的研究，它为研究核-线粒体相互作用提供了一个可替代且互补的模型，可用于补充芽殖酵母酿酒酵母的研究。粟酒裂殖酵母与哺乳动物细胞在呼吸生理学和线粒体基因组结构方面都有显著的相似之处。本技术综述简要列出了一些通用和特定的特性，这些特性有助于在开始将裂殖酵母用作线粒体研究的模型系统时了解。此外，还给出了细胞生长和基因技术方面的建议，介绍了破坏参与呼吸作用的基因的技巧，并提供了一种基本的差速离心方案，用于分离适合多种应用（如亚分级分离和体外导入）的纯化线粒体。
