Dabeva M D, Todorov B N, Khadzhiolova A A
Biokhimiia. 1976 Mar;41(3):458-68.
A method is described for the isolation of pure liver nuclei with minimal cytoplasmic contaminants, loss of nuclear RNA and degradation of nuclear RNA. The RNA components are extracted in three distinct fractions by subsequent treatment with phenol at 4 degrees, 50 degrees and 85 degrees C. The total and 14C-orotate labelled RNA components in the three nuclear RNA fractions are characterized by nucleotide composition, poly(A)-RNA content and agar-gel electrophoresis. The results show that the RNA in three fractions correspond to the nucleosol, nucleolus and chromatin compartments of the nucleus. The nuclear HnRNA components are exclusively in the 85 degrees C RNA. Nuclear ribosomal RNA is extracted in the 4 degrees C and 50 degrees C RNA fractions. These two nuclear RNA fractions are distinct in constituent pre-rRNA species and the rate of labelling of their rRNA components. The amount of the pre-rRNA and rRNA species is determined. The results show that the nucleolus-nucleosol and nucleosol-cytoplasm transitions of ribosomal subparticles are markedly slower processes than the preceeding steps of ribosome biogenesis.
本文描述了一种分离纯肝细胞核的方法，该方法可使细胞质污染物最少，核RNA损失最小且核RNA不发生降解。通过在4℃、50℃和85℃下依次用苯酚处理，将RNA成分提取为三个不同的组分。通过核苷酸组成、聚（A）-RNA含量和琼脂凝胶电泳对三个核RNA组分中的总RNA和14C-乳清酸标记的RNA成分进行表征。结果表明，三个组分中的RNA分别对应于细胞核的核溶质、核仁和染色质区室。核不均一核RNA（HnRNA）成分仅存在于85℃的RNA中。核糖体RNA在4℃和50℃的RNA组分中被提取。这两个核RNA组分在组成前体rRNA种类及其rRNA成分的标记速率方面有所不同。测定了前体rRNA和rRNA种类的量。结果表明，核糖体亚颗粒的核仁-核溶质和核溶质-细胞质转变过程明显比核糖体生物发生的先前步骤慢。
