Niessing J
Eur J Biochem. 1978 Nov 15;91(2):587-98. doi: 10.1111/j.1432-1033.1978.tb12712.x.
A procedure is described for the chromatographic analysis of RNA under fully denaturing conditions on cross-linked Sepharose. Chromatography of DNA . RNA hybrids, poly(C) . poly(G) hybrids and complexes of poly(C) . hnRNA on Sepharose CL in pure formamide at 46 degrees C leads to denaturation and strand separation of the hybrid structures. Using this procedure, nuclear RNA from duck immature red blood cells was resolved according to molecular size and assayed for the presence of globin mRNA sequences by hybridization with complementary DNA. Two size classes of putative globin mRNA precursor molecules were detected at an elution position corresponding to 14--18 S and 23--28 S. As judged from chromatographic analysis on poly(U)-Sepharose, about 70% of the 14--18-S globin precursor RNA is polyadenylated while only 11% of the putative 23--28-S precursor RNA has a poly(A) tract. Inhibition of transcription by actinomycin D and pulse-chase experiments indicate a half-life of less than 7.5 min for these precursor RNA species.
本文描述了一种在交联琼脂糖上进行完全变性条件下RNA色谱分析的方法。DNA.RNA杂交体、聚（C）.聚（G）杂交体以及聚（C）.不均一核RNA复合物在46℃纯甲酰胺中于琼脂糖CL上进行色谱分析，会导致杂交结构的变性和链分离。使用该方法，根据分子大小对鸭未成熟红细胞的核RNA进行分离，并通过与互补DNA杂交检测珠蛋白mRNA序列的存在。在对应于14 - 18 S和23 - 28 S的洗脱位置检测到两类假定的珠蛋白mRNA前体分子。从聚（U）-琼脂糖色谱分析判断，14 - 18 S珠蛋白前体RNA约70%是聚腺苷酸化的，而假定的23 - 28 S前体RNA只有11%有聚（A）尾。放线菌素D抑制转录以及脉冲追踪实验表明这些前体RNA种类的半衰期小于7.5分钟。
