Hunger H D, Grützmann H, Speer A, Hanke R, Hahn V, Coutelle C
Acta Biol Med Ger. 1979;38(11-12):1555-63.
Different fractions of cellular RNA from erythroid enriched bone marrow cells of the rabbit, extracted by the temperature fractionation method, were investigated by hybridization to globin cDNA. 97.4% of all globin sequences were found in the 4 degrees C franction (cytoplasmic RNA) 0.11% are in the 40 degrees / 50 degrees C fraction and 2.47% in the 65 degrees C and 85 degrees C franctions (pre-mRNA). This shows a substantial purification of the pre-mRNA fractions from cytoplasmic mRNA. 33% of the globin sequences in the 65 degrees C and 85 degrees C fractions are polyadenylated. The poly(A)+-RNA from the 65 degrees C and 85 degrees C fractions separated in a formamide sucrose gradient showed a clear hybridization to globin cDNA in the region between 9S and 28S and around 4S. In a control experiment in which RNA from baby hamster kidney cells (BHK) was mixed with globin mRNA and separated in the same manner hybridization was observed at the 9S position of the gradient only.
采用温度分级分离法从兔富含红系细胞的骨髓细胞中提取细胞RNA的不同级分,通过与珠蛋白cDNA杂交进行研究。发现所有珠蛋白序列的97.4%存在于4℃级分(细胞质RNA)中,0.11%存在于40℃/50℃级分中,2.47%存在于65℃和85℃级分(前体mRNA)中。这表明前体mRNA级分已从细胞质mRNA中得到大量纯化。65℃和85℃级分中33%的珠蛋白序列是聚腺苷酸化的。在甲酰胺蔗糖梯度中分离的65℃和85℃级分的聚(A)+RNA在9S至28S区域以及约4S处与珠蛋白cDNA呈现明显杂交。在一个对照实验中,将幼仓鼠肾细胞(BHK)的RNA与珠蛋白mRNA混合并以相同方式分离,仅在梯度的9S位置观察到杂交。