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通过限制性核糖核酸酶T2水解法分离和分析大鼠肝脏5S RNA的(Gp)nXp序列

Isolation and anaylsis of (Gp)nXp sequences of rat liver 5S RNA by means of restricted ribonucleas T2 hydrolysis.

作者信息

Willems R, Avdonina T, Lund A, Kisselev L L

出版信息

Nucleic Acids Res. 1974 Sep;1(9):1201-5. doi: 10.1093/nar/1.9.1201.

Abstract

Essentual difficulties arise when base number in oligoguanylic blocks and location of these blocks along the polynucleotide chain need to be determined in the course of determination of the nucleotide sequences in ribonucleic acids. To overcome this difficulty it is suggested to take advantage of a recently discovered resistance of phosphodiester bond between kethoxalated G and its 3'-neighbour against T(2) RNase hydrolysis 1,2. The approach is illustrated by analysis of 5S RNA from rat liver. Sequences of general formula (Gp)(n)Xp were isolated from T(2) RNase hydrolysate of 5 S RNA rapidly and quantitatively. The information obtained greatly facilitates the whole procedure of sequencing. It is expected that the method proposed would be effective for analysis of 5 S and 4 S RNA and for highmolecular weight fragments of ribosomal and viral RNAs.

摘要

在测定核糖核酸中的核苷酸序列过程中,当需要确定寡聚鸟苷酸片段中的碱基数目及其在多核苷酸链上的位置时,就会出现一些基本困难。为克服这一困难,建议利用最近发现的乙二醛化的鸟苷与其3'-邻位之间的磷酸二酯键对T(2)核糖核酸酶水解具有抗性这一特性。通过对大鼠肝脏5S RNA的分析来说明该方法。从5S RNA的T(2)核糖核酸酶水解产物中快速且定量地分离出通式为(Gp)(n)Xp的序列。所获得的信息极大地促进了整个测序过程。预计所提出的方法对5S和4S RNA以及核糖体和病毒RNA的高分子量片段的分析将是有效的。

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