Karpinska B, Karpinski S, Hällgren J E
Department of Forest Genetics and Plant Physiology, Faculty of Forestry, Swedish University of Agricultural Sciences, Umeå, Sweden.
Curr Genet. 1995 Oct;28(5):423-8. doi: 10.1007/BF00310810.
The nucleotide sequence of the region encoding NADH dehydrogenase subunit 3 and ribosomal protein S12 from Pinus sylvestris (L.) mitochondrial DNA (mtDNA) has been determined. A sequence comparison of this region with six individual cDNA clones prepared by RT-PCR revealed 35 C-to-T differences, showing the occurrence of RNA editing. All but one of these alterations in mRNA sequence change codon identities to specify amino acid better conserved in evolution. Most of these modifications take place within the nad3 gene changing 20% of the amino-acid sequence, which is much more than in angiosperms. Of six cDNA clones investigated, four clones of nad3 were differentially edited, but the editing of the rps12 sequences was identical. As in angiosperms, the two genes are separated by a short sequence of 52 bp, which is not edited. Two transcripts of about 0.9 kb and 1.2 kb, each encoding both proteins, have been detected by Northern hybridisation. The hybridisation of nad3 and rps12 probes with pine mtDNA digested with different restriction enzymes indicates that both genes are present in a single copy in pine mtDNA. The analysis of PCR amplification products with gene-specific primers shows a conserved order of these genes in a wide range of gymnosperms.
已确定欧洲赤松线粒体DNA(mtDNA)中编码NADH脱氢酶亚基3和核糖体蛋白S12区域的核苷酸序列。将该区域与通过RT-PCR制备的六个单独的cDNA克隆进行序列比较,发现有35个C到T的差异,表明存在RNA编辑。mRNA序列中的这些改变除了一个之外,其余都改变了密码子的同一性,以指定在进化中更保守的氨基酸。这些修饰大多发生在nad3基因内,改变了20%的氨基酸序列,这比被子植物中的要多得多。在所研究的六个cDNA克隆中,nad3的四个克隆有差异编辑,但rps12序列的编辑是相同的。与被子植物一样,这两个基因被一段52 bp的短序列隔开,该序列未被编辑。通过Northern杂交检测到了两个约0.9 kb和1.2 kb的转录本,每个转录本都编码这两种蛋白质。nad3和rps12探针与用不同限制酶消化的松树mtDNA杂交表明,这两个基因在松树mtDNA中均以单拷贝形式存在。用基因特异性引物对PCR扩增产物进行分析表明,在广泛的裸子植物中,这些基因具有保守的顺序。