Komiya H, Shimizu N, Kawakami M, Takemura S
J Biochem. 1980 Nov;88(5):1449-56. doi: 10.1093/oxfordjournals.jbchem.a133114.
By chromatography on columns of DEAE-Sephadex A-50 and Sephadex G-100, and electrophoresis on polyacrylamide gel, 5S rRNA was purified from a low-molecular-weight RNA fraction extracted from the total tissues of Lingula anatina. Complete digests of the 5S rRNA with RNase T1 [EC 3.1.4.8] and pancreatic RNase [EC 3.1.4.22] were sequenced by conventional column chromatography procedures. The nucleotide sequence of this RNA was determined mainly by a chemical method for sequencing the RNA 3' end-labeled with 32P (1), with the complement of the oligonucleotide catalog obtained by the complete RNase digestions of the RNA. By comparing the sequences of several invertebrate, vertebrate, and Chlorella 5S rRNAs, a phylogenic tree of the rRNAs was constructed and the time of divergence of Lingula was estimated.
通过在DEAE - 葡聚糖凝胶A - 50和葡聚糖凝胶G - 100柱上进行层析,以及在聚丙烯酰胺凝胶上进行电泳,从鸭嘴海豆芽总组织中提取的低分子量RNA组分中纯化出了5S rRNA。用核糖核酸酶T1[EC 3.1.4.8]和胰核糖核酸酶[EC 3.1.4.22]对5S rRNA进行完全消化,并通过常规柱层析程序对其进行测序。该RNA的核苷酸序列主要通过一种化学方法确定,该方法用于对用32P进行3'末端标记的RNA进行测序(1),并结合通过RNA的完全核糖核酸酶消化获得的寡核苷酸目录的互补序列。通过比较几种无脊椎动物、脊椎动物和小球藻5S rRNA的序列,构建了rRNA的系统发育树,并估计了鸭嘴海豆芽的分化时间。