Childs G, Levy S, Kedes L H
Biochemistry. 1979 Jan 9;18(1):208-13. doi: 10.1021/bi00568a032.
We describe a rapid and simple method for the purification of biologically active messenger RNAs. The method allows the isolation in a few hours of specific mRNAs from either whole cell or polysomal RNA even if the RNA represents less than 1% of the starting molecules. We used, as a model, cloned sea urchin (Strongylocentrotus purpuratus) histone gene fragments linked to cellulose by the method of B. E. Noyes & G. R. Stark (1975) Cell 5, 301--310) as hybridization probes to isolate specific histone mRNAs from whole cell and polysomal RNA extracts. RNAs isolated in this manner maintain their biological activity, serving as templates for histone proteins in a wheat-germ, cell-free protein translation system. In addition, radiolabeled histone-specific RNA purified from cleavage stage sea urchin embryos, pulse labeled for short periods of time and analyzed on denaturing polyacrylamide gels, was the same size as mature histone mRNA's.
我们描述了一种快速简便的纯化具有生物活性信使核糖核酸（mRNA）的方法。该方法能在数小时内从全细胞或多聚核糖体RNA中分离出特定的mRNA，即便该RNA在起始分子中所占比例不到1%。我们以通过B.E.诺伊斯和G.R.斯塔克（1975年，《细胞》5卷，301 - 310页）的方法与纤维素相连的克隆海胆（紫球海胆）组蛋白基因片段作为杂交探针，从全细胞和多聚核糖体RNA提取物中分离特定的组蛋白mRNA。以这种方式分离的RNA保持其生物活性，可在小麦胚芽无细胞蛋白质翻译系统中作为组蛋白的模板。此外，从处于卵裂期的海胆胚胎中纯化的经短时间脉冲标记并在变性聚丙烯酰胺凝胶上分析的放射性标记组蛋白特异性RNA，其大小与成熟组蛋白mRNA相同。
