关于人转运核糖核酸的研究。I. 胎盘和肝脏转运核糖核酸的快速大规模分离及部分分级分离
Studies on human tRNA. I. The rapid, large scale isolation and partial fractionation of placenta and liver tRNA.
作者信息
Roe B A
出版信息
Nucleic Acids Res. 1975 Jan;2(1):21-42. doi: 10.1093/nar/2.1.21.
Abstract
A procedure for the large scale isolation of mammalian tRNA has been applied to the isolation of several grams of human liver, human placenta, rabbit liver and rat liver tRNA. This procedure entails an initial grinding of the tissue in phenol-sodium acetate at acidic pH, followed by DEAE cellulose chromatography. Procedures are also described for analysis of the purified tRNA on the basis of size, using controlled pore glass bean columns. In addition, the acceptor activity of isolated tRNAs has been determined using both the heterologous and homologous synthetases. The chromatographic profile of individual isoaccepting species using BD cellulose chromatography is shown and the 3' terminal nucleoside content was also determined. The methods described now make it feasible for large scale studies of mammalian tRNA enabling us to better understand the relationships between the structure of mammalian tRNA and its many diversified functions.
摘要
一种大规模分离哺乳动物tRNA的方法已被应用于从几克人类肝脏、人类胎盘、兔肝脏和大鼠肝脏中分离tRNA。该方法首先在酸性pH条件下于苯酚 - 醋酸钠中研磨组织，随后进行DEAE纤维素色谱分离。还介绍了使用可控孔径玻璃珠柱基于大小分析纯化tRNA的方法。此外，已使用异源和同源合成酶测定了分离的tRNA的受体活性。展示了使用BD纤维素色谱法对各个同功受体种类的色谱图谱，并且还测定了3'末端核苷含量。现在所描述的方法使得对哺乳动物tRNA进行大规模研究成为可能，从而使我们能够更好地理解哺乳动物tRNA的结构与其多种多样化功能之间的关系。
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