Hew C L, Penner P E
Can J Biochem. 1979 Jul;57(7):1030-5. doi: 10.1139/o79-129.
The induction of rat liver zinc-thioneins mRNA was studied in a wheat germ cell-free translation system. Liver poly A rich polysomal RNA was isolated from rats which had been injected with zinc sulfate 5 h previously. These RNA preparations stimulated the incorporation of [35S]cystine into trichloroacetic acid insoluble proteins when assayed in the cell-free synthetic system. The translation products were characterized by Sephadex G-75 chromatography in 8 M urea--50 mM beta-mercaptoethanol, by disc gel electrophoresis in 4 M area--Tris-glycine buffer (pH 9.2), and by peptide fingerprinting with pepsin. These results were identical with authentic rat liver zinc-thioneins. The zinc-thioneins mRNA activity in the control rats, however, was minimal. The stimulation in zinc-thioneins synthesis observed in the cell-free synthesis was similar to the increased synthesis of these polypeptides in vivo.
在小麦胚无细胞翻译系统中研究了大鼠肝脏锌硫蛋白mRNA的诱导情况。从5小时前注射过硫酸锌的大鼠中分离出富含多聚腺苷酸的肝脏多聚核糖体RNA。当在无细胞合成系统中进行测定时，这些RNA制剂能刺激[35S]胱氨酸掺入三氯乙酸不溶性蛋白质中。通过在8M尿素-50mMβ-巯基乙醇中进行Sephadex G-75层析、在4M脲- Tris-甘氨酸缓冲液（pH 9.2）中进行圆盘凝胶电泳以及用胃蛋白酶进行肽指纹图谱分析来对翻译产物进行表征。这些结果与大鼠肝脏锌硫蛋白的标准品一致。然而，对照大鼠中锌硫蛋白mRNA的活性极低。在无细胞合成中观察到的锌硫蛋白合成的刺激与这些多肽在体内合成的增加相似。
