Brown G D, Peluso R W, Moyer S A, Moyer R W
J Biol Chem. 1983 Dec 10;258(23):14309-14.
A rapid, simple procedure is described for the preparation of cell-free extracts of both uninfected and virus-infected pig kidney and baby hamster kidney cells which are very active in the in vitro translation of not only endogenous mRNA, but also of exogenous mRNA added to extracts that had been depleted of endogenous mRNA by treatment with micrococcal nuclease. This procedure appears to be adaptable with only minor variations to many eukaryotic cell lines and should greatly facilitate in vitro molecular studies of protein synthesis and gene regulation at the level of translation.
本文描述了一种快速、简单的方法,用于制备未感染和病毒感染的猪肾细胞及幼仓鼠肾细胞的无细胞提取物。这些提取物不仅在体外翻译内源性mRNA方面非常活跃,而且在用微球菌核酸酶处理以耗尽内源性mRNA的提取物中添加外源性mRNA后,也能高效翻译。该方法似乎只需稍作改动就能适用于许多真核细胞系,并且应能极大地促进在翻译水平上进行蛋白质合成和基因调控的体外分子研究。