Nagley P, Clayton D A
Nucleic Acids Res. 1980 Jul 11;8(13):2947-65. doi: 10.1093/nar/8.13.2947.
The map positions in mouse mitochondrial DNA of the two ribosomal RNA genes and adjacent genes coding several small transcripts have been determined precisely by application of a procedure in which DNA-RNA hybrids have been subjected to digestion by S1 nuclease under conditions of varying severity. Digestion of the DNA-RNA hybrids with S1 nuclease yielded a series of species which were shown to contain ribosomal RNA molecules together with adjacent transcripts hybridized conjointly to a continuous segment of mitochondrial DNA. There is one small transcript about 60 bases long whose gene adjoins the sequences coding the 5'-end of the small ribosomal RNA (950 bases) and which lies approximately 200 nucleotides from the D-loop origin of heavy strand mitochondrial DNA synthesis. An 80-base transcript lies between the small and large ribosomal RNA genes, and genes for two further short transcript (each about 80 bases in length) abut the sequences coding the 3'-end of the large ribosomal RNA (approximately 1500 bases). The ability to isolate a discrete DNA-RNA hybrid species approximately 2700 base pairs in length containing all these transcripts suggests that there can be few nucleotides in this region of mouse mitochondrial DNA which are not represented as stable RNA species.
通过应用一种程序,在不同严格程度的条件下用S1核酸酶消化DNA - RNA杂种,已精确确定了小鼠线粒体DNA中两个核糖体RNA基因以及编码几个小转录本的相邻基因的图谱位置。用S1核酸酶消化DNA - RNA杂种产生了一系列片段,这些片段显示包含核糖体RNA分子以及与线粒体DNA连续片段联合杂交的相邻转录本。有一个约60个碱基长的小转录本,其基因邻接编码小核糖体RNA(950个碱基)5'端的序列,并且它距离重链线粒体DNA合成的D环起始点约200个核苷酸。一个80个碱基的转录本位于小核糖体RNA基因和大核糖体RNA基因之间,另外两个短转录本(每个约80个碱基长)的基因邻接编码大核糖体RNA(约1500个碱基)3'端的序列。能够分离出一个长度约为2700个碱基对、包含所有这些转录本的离散DNA - RNA杂种片段,这表明在小鼠线粒体DNA的这个区域中,未作为稳定RNA片段存在的核苷酸很少。