[牛乳腺tRNA2Leu的一级结构。T1核糖核酸酶水解产物的寡核苷酸。完整核苷酸序列的重建]

[Primary structure of tRNA2Leu of bovine mammary gland. Oligonucleotides of T1-RNAse hydrolysate. Reconstruction of a total nucleotide sequence].

作者信息

Tukalo M A, Vasil'eva I G, Matsuka G Kh, Vlasov V B

出版信息

Bioorg Khim. 1984 Jan;10(1):58-67.

Abstract

The oligonucleotides obtained by digestion of tRNA2Leu from cow mammary gland with T1 RNase were separated by micro-column chromatography on DEAE-cellulose in 7 M urea at pH 7,5 and 3,7, and in addition on Dowex 1 x 2. The digest consisted of 18 individual components, the larger being a tridecanucleotide. Micro-column chromatography of nucleotides on anion-exchanger AG 1 x 8 and nucleosides on Aminex A-6 was used to determine the base composition of the oligonucleotides. The oligonucleotide structure was established using terminal analysis, hydrolysis by pancreatic and U2-RNases and incomplete hydrolysis by snake venom phosphodiesterases. The total primary structure of tRNA2Leu was derived from overlapping fragments isolated after its complete hydrolysis with pancreatic and T1 RNase and using data obtained on S1-nuclease digestion of tRNA. The methods of rapid gel-sequencing were also employed for checking the nucleotide sequence of tRNA2Leu from cow mammary gland.

摘要

用T1核糖核酸酶消化牛乳腺中的tRNA2Leu所得到的寡核苷酸，通过在pH 7.5和3.7的7M尿素中于DEAE - 纤维素上进行微柱层析，以及另外在Dowex 1×2上进行分离。消化产物由18个单独的组分组成，较大的是一个十三聚核苷酸。利用阴离子交换剂AG 1×8上的核苷酸微柱层析和Aminex A - 6上的核苷微柱层析来测定寡核苷酸的碱基组成。使用末端分析、胰核糖核酸酶和U2 - 核糖核酸酶水解以及蛇毒磷酸二酯酶不完全水解来确定寡核苷酸结构。tRNA2Leu的完整一级结构是从用胰核糖核酸酶和T1核糖核酸酶完全水解后分离出的重叠片段推导出来的，并利用了tRNA经S1核酸酶消化所获得的数据。快速凝胶测序方法也被用于检查牛乳腺tRNA2Leu的核苷酸序列。

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Nucleic Acids Res. 1996 Apr 15;24(8):1489-96. doi: 10.1093/nar/24.8.1489.

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