Eladari M E, Hampe A, Galibert F
Biochimie. 1979;61(9):1073-80. doi: 10.1016/s0300-9084(80)80261-6.
The primary structure of 17S and 25S ribosomal RNAs from Saccharomyces cerevisiae has been analysed by two-dimensional fractionation of T1 oligonucleotides. This method consists of an electrophoresis at pH 3.5 followed by a homochromatography on DEAE-cellulose plates. After the second dimension, the large T1 oligonucleotides were hydrolyzed by pancreatic RNAse, followed by alkaline hydrolysis of the pancreatic products. By fractionating a mixture of tritiated HeLa cell ribosomal RNAs and 32 P yeast cell ribosomal RNAs, two autoradiographs were obtained; one corresponding to the 32P labelled material and the other to the tritiated labelled material. By superposition of the two autoradiographs, the mobility of the various T1 oligonucleotides can be accurately compared and it is shown that yeast 17S rRNA and human 18S rRNA have in common 5 large oligonucleotides and that yeast 25S rRNA and human 28S rRNA have 4 identical oligonucleotides.
通过T1寡核苷酸的二维分级分离,分析了酿酒酵母17S和25S核糖体RNA的一级结构。该方法包括在pH 3.5下进行电泳,然后在DEAE-纤维素板上进行同系层析。在第二维之后,大的T1寡核苷酸用胰核糖核酸酶水解,随后对胰产物进行碱性水解。通过对氚标记的HeLa细胞核糖体RNA和32P酵母细胞核糖体RNA的混合物进行分级分离,获得了两张放射自显影片;一张对应于32P标记的物质,另一张对应于氚标记的物质。通过叠加这两张放射自显影片,可以准确比较各种T1寡核苷酸的迁移率,结果表明酵母17S rRNA和人18S rRNA共有5个大的寡核苷酸,酵母25S rRNA和人28S rRNA有4个相同的寡核苷酸。