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哺乳动物细胞中核糖体RNA成熟的指纹图谱研究。

Fingerprinting studies of the maturation of ribosomal RNA in mammalian cells.

作者信息

Galibert F, Tiollais P, Eladari M E

出版信息

Eur J Biochem. 1975 Jun 16;55(1):239-45. doi: 10.1111/j.1432-1033.1975.tb02156.x.

Abstract

32P-labelled ribosomal RNA of L 5178 Y cells (a mouse cell line) was digested with T1 ribonuclease and fingerprinted by electrophoresis at pH 3.5 on cellulose acetate and homochromatography on DEAE-cellulose thin-layer plate. From this, it can be concluded that 18-S and 28-S RNA have different and characteristic fingerprints and that the number, the size and the frequency of the large T1 oligonucleotides demonstrate that the guanylic residues are randomly interspaced along the molecule. Using a double-labelling technique with 32P-labelled 45-S RNA and 14C-labelled 18-S RNA or 28-S RNA, long T1 oligonucleotides of the 45-S RNA can be divided into three classes: (a) those which are lost during the transition, (b) those which are present in the 18-S RNA and (c) those which are present in the 28-S RNA. These results provide direct evidence for the existence of one common precursor for the two mature ribosomal RNAs. The comparison of the fingerprints of T1-ribonuclease-digested 47-S, 45-S and 41-S RNA shows that the 47-S and 41-S RNA have a characteristic ribosomal pattern. Finally the size, the number and the mobility of the oligonucleotides present in the different RNA precursors but absent from the mature RNA demonstrate that the non-conversed RNA pieces do not have a monotonous and repetitive sequence.

摘要

用T1核糖核酸酶消化L 5178 Y细胞(一种小鼠细胞系)的32P标记核糖体RNA,并通过在pH 3.5条件下于醋酸纤维素上进行电泳以及在DEAE - 纤维素薄板上进行同系层析进行指纹图谱分析。由此可以得出结论,18 - S和28 - S RNA具有不同的特征性指纹图谱,并且大的T1寡核苷酸的数量、大小和频率表明鸟苷酸残基沿分子随机间隔分布。使用32P标记的45 - S RNA和14C标记的18 - S RNA或28 - S RNA的双标记技术,45 - S RNA的长T1寡核苷酸可分为三类:(a)在转变过程中丢失的那些;(b)存在于18 - S RNA中的那些;(c)存在于28 - S RNA中的那些。这些结果为两种成熟核糖体RNA存在一个共同前体提供了直接证据。对T1核糖核酸酶消化的47 - S、45 - S和41 - S RNA的指纹图谱比较表明,47 - S和41 - S RNA具有特征性的核糖体模式。最后,存在于不同RNA前体但不存在于成熟RNA中的寡核苷酸的大小、数量和迁移率表明,未转化的RNA片段不具有单调且重复的序列。

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