Xue Y, Collin S, Davies D R, Thomas C M
AFRC Institute of Plant Science Research, John Innes Research Centre, Norwich, UK.
Plant Mol Biol. 1994 Apr;25(1):91-103. doi: 10.1007/BF00024201.
As part of a strategy to define differences in genome organization and expression between cytoplasmic male-sterile (CMS) and male-fertile (MF) sugar-beet mitochondria, cDNA libraries from both mitochondrial genotypes were constructed. Preliminary screening with ribosomal RNA gene probes identified candidate cDNA clones corresponding to structural genes. In addition, reciprocal hybridization experiments were performed using labelled first-strand cDNA to identify uniquely transcribed sequences. One cDNA clone (pYC700) is unique to CMS mitochondria and is located upstream of the F0F1-ATPase subunit 6 gene (atp6). Another cDNA clone (pYC130), when used as a probe in northern hybridization analysis, revealed novel transcript profiles in CMS sugar-beet mitochondria. Sequence analysis of this cDNA showed strong homology with the F0F1-ATPase subunit alpha (atpA) coding sequences from several higher plants. The atp6 and atpA loci from each genotype were cloned and the genomic organization, DNA sequence and transcription of each locus was studied. Differences in the transcript profiles of each gene are a consequence of genomic rearrangements 5' to the coding sequence.
作为确定细胞质雄性不育（CMS）和雄性可育（MF）甜菜线粒体基因组组织和表达差异策略的一部分，构建了两种线粒体基因型的cDNA文库。用核糖体RNA基因探针进行初步筛选，鉴定出与结构基因对应的候选cDNA克隆。此外，使用标记的第一链cDNA进行了反向杂交实验，以鉴定独特转录的序列。一个cDNA克隆（pYC700）是CMS线粒体特有的，位于F0F1 - ATP酶亚基6基因（atp6）的上游。另一个cDNA克隆（pYC130）在Northern杂交分析中用作探针时，揭示了CMS甜菜线粒体中的新转录本谱。该cDNA的序列分析显示与几种高等植物的F0F1 - ATP酶亚基α（atpA）编码序列具有高度同源性。克隆了每种基因型的atp6和atpA基因座，并研究了每个基因座的基因组组织、DNA序列和转录情况。每个基因转录本谱的差异是编码序列5'端基因组重排的结果。
