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酵母中线粒体突变体的筛选。

Screens for mitochondrial mutants in yeast.

作者信息

Foury Françoise

机构信息

Unité de Biochimie Physiologique, Louvain-la-Neuve, Belgium.

出版信息

Methods Mol Biol. 2007;372:167-76. doi: 10.1007/978-1-59745-365-3_12.

Abstract

We describe methods that allow isolation, identification, and counting of mitochondrial mutants that are resistant to antibiotics (ant(R)) or respiratory deficient (rho-). (1) Analysis of diploid and meiotic progenies generated in crosses between mutants and tester strains allows distinguishing nuclear from mitochondrial mutants, for either antibiotic resistance or respiratory deficiency. (2) The mutation rate of mitochondrial deoxyribonucleic acid (mtDNA) can be estimated from the average frequency of ant(R) mutants produced in a large number of independent clones. (3) The frequency of retention of mtDNA fragments in rho- genomes accumulating in nuclear respiratory-deficient mutants can be determined by a genetic test based on the ability of these rho- genomes to restore cellular growth on glycerol in crosses with selected mutants bearing punctual mutations in their mtDNA (mit-).

摘要

我们描述了一些方法,可用于分离、鉴定和计数对抗生素具有抗性(ant(R))或呼吸缺陷(rho-)的线粒体突变体。(1)分析突变体与测试菌株杂交产生的二倍体和减数分裂后代,可区分核突变体和线粒体突变体,无论是抗生素抗性还是呼吸缺陷方面。(2)线粒体脱氧核糖核酸(mtDNA)的突变率可根据大量独立克隆中产生的ant(R)突变体的平均频率来估计。(3)核呼吸缺陷突变体中积累的rho-基因组中线粒体DNA片段的保留频率,可通过基于这些rho-基因组在与线粒体DNA中存在点突变(mit-)的选定突变体杂交时恢复甘油上细胞生长能力的遗传测试来确定。

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