Fractionation of rat liver tRNA by reversed-phase high performance liquid chromatography: isolation of Iso-tRNAs(Pro).

D Kanduc
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引用次数: 7

Abstract

This paper illustrates the fractionation of cytoplasmic transfer ribonucleic acid from rat liver by reversed-phase high performance liquid chromatography using a gradient of acetonitrile/ammonium acetate. The procedure is fast, highly reproducible, and gives an excellent resolution of the numerous tRNA population: about 50 peaks with area peak percentages ranging from 0.001 to 5 can be monitored. Uncharged tRNA preparations exhibited a chromatographic profile different from aminoacylated tRNA, thus suggesting a possible strategy to distinguish between aminoacylated and nonacylated tRNA species. Moreover, a first approach to map the HPLC peaks was attempted by chromatographing preparations of tRNA which had been aminoacylated with individual 3H-labeled aminoacids. Here is reported the case of tRNA(Pro), which gave three well separated radioactive peaks, most likely corresponding to tRNA(Pro) isoacceptor species.

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反相高效液相色谱法分离大鼠肝脏tRNA:分离iso -tRNA (Pro)。
本文介绍了用乙腈/乙酸铵梯度反相高效液相色谱法分离大鼠肝脏细胞质转移核糖核酸的方法。该方法快速,重复性高,并提供了大量tRNA种群的极好分辨率:可以监测约50个峰,面积峰百分比范围为0.001至5%。未带电tRNA的制备表现出与氨基酰化tRNA不同的色谱特征,从而提出了区分氨基酰化和非酰化tRNA的可能策略。此外,通过用单个3h标记的氨基酸进行氨基酰化的tRNA的色谱制备,尝试了绘制HPLC峰的第一种方法。本文报道了tRNA(Pro)的例子,它给出了三个分离良好的放射性峰,很可能对应于tRNA(Pro)的同受体种类。
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