Lichtenstein A V, Alechina R P, Shapot V S
Biochem J. 1975 Jun;147(3):447-56. doi: 10.1042/bj1470447.
A novel method of RNA fractionation has been developed. Nuclear and cytoplasmic rat liver RNA species were fractionated as constituents of corresponding ribonucleoprotein particles, which were previously adsorbed on a Celite-column by their protein component. The fractionation is based on a dissociation of the particles (linear concentration gradient of LiCl and urea with subsequent temperature gradient), which results in a gradual release of the RNA molecules from ribonucleoprotein complexes. Thus the fractionation is in accordance with the tightness of the RNA-protein bonds. A gradient elution of RNA from a nucleoprotein-Celite column permitted fractionation of both ribosomal and rapidly labelled non-ribosomal RNA. The latter, both nuclear and cytoplasmic, could be separated by chromatography on nucleoprotein-Celite columns into two main fractions (components I and V). In cytoplasmic RNA components I and V presumably correspond to mlRNA (messenger-like RNA of free cytoplasmic particles) and mRNA (template RNA associated with ribosomes) respectively.
一种新的RNA分级分离方法已经被开发出来。大鼠肝脏的核RNA和细胞质RNA种类作为相应核糖核蛋白颗粒的组成成分进行分级分离,这些核糖核蛋白颗粒先前通过其蛋白质成分吸附在硅藻土柱上。分级分离基于颗粒的解离(LiCl和尿素的线性浓度梯度以及随后的温度梯度),这导致RNA分子从核糖核蛋白复合物中逐渐释放出来。因此,分级分离与RNA-蛋白质键的紧密程度一致。从核蛋白-硅藻土柱上对RNA进行梯度洗脱,可以对核糖体RNA和快速标记的非核糖体RNA进行分级分离。后者,无论是核内的还是细胞质的,都可以通过在核蛋白-硅藻土柱上进行色谱分离,分为两个主要部分(组分I和V)。在细胞质RNA中,组分I和V可能分别对应于mlRNA(游离细胞质颗粒的类信使RNA)和mRNA(与核糖体相关的模板RNA)。