Dieckmann C L, Pape L K, Tzagoloff A
Proc Natl Acad Sci U S A. 1982 Mar;79(6):1805-9. doi: 10.1073/pnas.79.6.1805.
Nuclear pet mutants of Saccharomyces cerevisiae deficient in mitochondrial respiration have been studied genetically and biochemically. Seven noncomplementing mutations leading to a deficiency of mitochondrial cytochrome b have been assigned to a single complementation group (group 60). Examination of mitochondrial RNA by blot hybridization on diazobenzyloxymethyl-paper has revealed that group 60 mutants produce a large number of novel apocytochrome b transcripts not detected in wild-type yeast. The product of the gene affected in the mutants, therefore, appears to be required either for correct transcription or for processing of apocytochrome b premessenger RNA. The gene has been designated CBP1. A representative mutant from complementation group 60 (N5-26) has been transformed to respiratory competency with a recombinant plasmid pool consisting of random fragments of wild-type yeast nuclear DNA inserted into a vector capable of replicating in yeast and Escherichia coli. The complementation of the N5-26 mutation has been shown for a number of independent transformants to be due to the presence of plasmid DNA. The plasmid pG60/T10 was further characterized to have a nuclear DNA insert of 6.7 kilobase pairs. This plasmid complements the mutations of all group 60 mutants, thus confirming that it contains the CBP1 gene.
对线粒体呼吸功能缺陷的酿酒酵母核pet突变体进行了遗传学和生物化学研究。导致线粒体细胞色素b缺陷的七个非互补突变已被归入一个单一的互补群（第60组）。通过在重氮苄氧基甲基纸上进行印迹杂交来检测线粒体RNA，结果显示第60组突变体产生了大量在野生型酵母中未检测到的新型脱辅基细胞色素b转录本。因此，突变体中受影响基因的产物似乎是正确转录或脱辅基细胞色素b前体信使RNA加工所必需的。该基因被命名为CBP1。来自互补群60的一个代表性突变体（N5 - 26）已用一个重组质粒库转化为具有呼吸能力，该重组质粒库由插入到能够在酵母和大肠杆菌中复制的载体中的野生型酵母核DNA随机片段组成。已证明许多独立转化体对N5 - 26突变的互补是由于质粒DNA的存在。质粒pG60/T10经进一步鉴定，其核DNA插入片段为6.7千碱基对。该质粒可互补所有第60组突变体的突变，从而证实它包含CBP1基因。
