Bonnefoy Nathalie, Fox Thomas D
Centre de Génétique Moléculaire, UPR 2167, Gif-sur-Yvette, France.
Methods Mol Biol. 2007;372:153-66. doi: 10.1007/978-1-59745-365-3_11.
Saccharomyces cerevisiae is currently the only species in which genetic transformation of mitochondria can be used to generate a wide variety of defined alterations in mitochondrial deoxyribonucleic acid (mtDNA). DNA sequences can be delivered into yeast mitochondria by microprojectile bombardment (biolistic transformation) and subsequently incorporated into mtDNA by the highly active homologous recombination machinery present in the organelle. Although transformation frequencies are relatively low, the availability of strong mitochondrial selectable markers for the yeast system, both natural and synthetic, makes the isolation of transformants routine. The strategies and procedures reviewed here allow the researcher to insert defined mutations into endogenous mitochondrial genes and to insert new genes into mtDNA. These methods provide powerful in vivo tools for the study of mitochondrial biology.
酿酒酵母是目前唯一一种可利用线粒体基因转化在其线粒体脱氧核糖核酸（mtDNA）中产生多种特定改变的物种。DNA序列可通过微粒轰击（生物弹道转化）导入酵母线粒体，随后通过该细胞器中存在的高活性同源重组机制整合到mtDNA中。尽管转化频率相对较低，但酵母系统中天然和合成的强大线粒体选择标记的可用性使得转化子的分离成为常规操作。本文综述的策略和程序使研究人员能够将特定突变插入内源性线粒体基因，并将新基因插入mtDNA。这些方法为线粒体生物学研究提供了强大的体内工具。
