Borelli T J, Wu J M
Biochem Int. 1985 Mar;10(3):517-24.
Polyadenylated RNA has been isolated from control and interferon-treated HL-60 cells by centrifugation through cesium chloride and oligo(dT)-cellulose column chromatography. The affinity column-purified RNA is poorly translated in the mRNA-dependent rabbit reticulocyte lysates but is an excellent template for in vitro protein synthesis using the wheat germ cell extracts. The discrepancy in the efficiency of HL-60 mRNA utilization in the two commonly used cell-free protein synthesizing systems is attributable to an inhibitory component present in the polyadenylated RNA. This contaminant is most likely double-stranded RNA based on (i) the ability of 2-aminopurine (3-5 mM) or high concentrations of penicillium chrysogenum double-stranded RNA (10-15 micrograms/ml) to overcome the inhibition exerted by the component, and (ii) the ability of the component to promote the enzymatic conversion of ATP into 2-5A by the highly purified rabbit reticulocyte 2-5A synthetase.
通过氯化铯离心和寡聚（dT）-纤维素柱层析，从对照及经干扰素处理的HL-60细胞中分离出了聚腺苷酸化RNA。亲和柱纯化的RNA在依赖mRNA的兔网织红细胞裂解物中翻译效率很低，但却是利用小麦胚芽细胞提取物进行体外蛋白质合成的极佳模板。在两种常用的无细胞蛋白质合成系统中，HL-60 mRNA利用效率的差异归因于聚腺苷酸化RNA中存在的一种抑制成分。基于以下两点，这种污染物很可能是双链RNA：（i）2-氨基嘌呤（3-5 mM）或高浓度的产黄青霉双链RNA（10-15微克/毫升）能够克服该成分施加的抑制作用；（ii）该成分能够通过高度纯化的兔网织红细胞2-5A合成酶促进ATP酶促转化为2-5A。
