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甜菜线粒体小环状DNA:高分辨率转录图谱绘制、转录本丰度及拷贝数测定

Sugarbeet minicircular mitochondrial DNAs: high-resolution transcript mapping, transcript abundance and copy number determination.

作者信息

Thomas C M

机构信息

Department of Applied Genetics, John Innes Centre for Plant Science Research, Norwich, UK.

出版信息

Mol Gen Genet. 1992 Sep;234(3):457-65. doi: 10.1007/BF00538706.

Abstract

Three minicircular mitochondrial DNAs have been studied to address several aspects of transcription in sugarbeet mitochondria. High-resolution transcript mapping experiments have shown that sequences at the 5' termini of minicircle transcripts are highly homologous and resemble sequences at the 5' termini of sugarbeet mainband mitochondrial genes (atpA, atp6). In addition, they show homology to transcript termini of mitochondrial genes from other dicotyledonous plants, suggesting they may function as promoter sequences. Conserved sequences, which most probably act as RNA processing signals, were also identified at the 3' termini of minicircle transcripts. An oligonucleotide probe to a 14 base conserved sequence was used to determine the relative copy numbers of the three minicircle components in male-fertile mitochondria. Copy numbers were roughly equivalent, suggesting minicircles are replicated and/or transmitted with nearly equal efficiency, at least in sugarbeet taproots. Mc.a and Mc.c transcript levels are equivalent, consistent with their template copy number, however; Mc.d transcript levels were significantly lower than expected, implicating additional factors such as promoter strength and/or transcript stability in determining transcript levels in sugarbeet mitochondria, as recently demonstrated in maize.

摘要

为了研究甜菜线粒体转录的几个方面,对三个小环状线粒体DNA进行了研究。高分辨率转录图谱实验表明,小环转录本5'末端的序列高度同源,并且与甜菜主带线粒体基因(atpA、atp6)5'末端的序列相似。此外,它们与其他双子叶植物线粒体基因的转录本末端具有同源性,表明它们可能作为启动子序列发挥作用。在小环转录本的3'末端也鉴定出了最有可能作为RNA加工信号的保守序列。使用针对14个碱基保守序列的寡核苷酸探针来确定雄性可育线粒体中三个小环成分的相对拷贝数。拷贝数大致相当,这表明小环至少在甜菜主根中以几乎相等的效率进行复制和/或传递。然而,Mc.a和Mc.c的转录水平相当,与其模板拷贝数一致;Mc.d的转录水平明显低于预期,这表明在确定甜菜线粒体转录水平时还涉及其他因素,如启动子强度和/或转录本稳定性,正如最近在玉米中所证明的那样。

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