Vedeler A, Pryme I F, Hesketh J E
Department of Biochemistry, University of Bergen, Norway.
Mol Cell Biochem. 1991 Feb 2;100(2):183-93. doi: 10.1007/BF00234167.
Polysomes from Krebs II ascites and 3T3 cells were separated into three populations by using a sequential extraction method. Free polysomes were released by using a combination of low salt (25 mM KCl) and NP-40 detergent in the lysis buffer. The cytoskeletal bound polysomes were subsequently released by raising the salt concentration to 130 mM and finally, polysomes bound to the membranes of the endoplasmic reticulum were extracted by the combined treatment with Triton X-100 and deoxycholate. The results presented here illustrate that the three polysome-containing fractions differ in many parameters such as polysome profiles, cytoskeletal components and phospholipid content. When polyA-containing mRNA was isolated from the three polysome fractions and translated in an in vitro system, some differences were observed in the patterns of proteins being synthesized.
通过一种连续提取方法，将来自克雷布斯II腹水癌细胞和3T3细胞的多核糖体分离成三个群体。在裂解缓冲液中使用低盐（25 mM KCl）和NP - 40去污剂的组合释放游离多核糖体。随后通过将盐浓度提高到130 mM释放细胞骨架结合的多核糖体，最后，通过用Triton X - 100和脱氧胆酸盐联合处理提取与内质网膜结合的多核糖体。此处呈现的结果表明，这三个含多核糖体的组分在许多参数上存在差异，如多核糖体图谱、细胞骨架成分和磷脂含量。当从这三个多核糖体组分中分离出含polyA的mRNA并在体外系统中进行翻译时，在蛋白质合成模式上观察到了一些差异。
