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以1M蔗糖垫超离心法制备大鼠肝聚核蛋白体(Rp),去Rp的上清中再经35～50％饱和度硫酸铵盐析和DEAE-52-氯化钠梯度柱层析制备核糖核酸酶天然抑制物(RⅠ)。紫外光谱和蔗糖梯度超离心分析及麦胚无细胞体系翻译活力测定表明制备过程中Rp基本保持完整,有较高的促进放射性氨基酸参入活力。RⅠ经部分纯化活力提高100倍以上,把它加入到麦胚体系,能进一步提高Rp及兔网织红细胞聚核蛋白体RNA的模板活力。
Rat hepatic polyribosomes (Rp) were prepared by ultracentrifugation using 1 M sucrose pad. The supernatant from Rp was further purified by ammonium sulfate salting-out with 35-50% saturation and DEAE-52-NaCl gradient column chromatography Natural inhibitor of ribonuclease (RI). Ultraviolet spectroscopy and sucrose gradient ultracentrifugation analysis and wheat germ cell-free system translational activity determination showed that during the preparation of Rp remained intact, there is a higher promotion of radioactive amino acids into the vitality. R I was partially purified more than 100 times more active, adding it to the wheat germ system, can further enhance the Rp and rabbit reticulocyte polyribonucleic RNA template activity.