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用甲基化白蛋白-硅藻土柱对大鼠组织中的核核糖核酸和细胞质核糖核酸进行分级分离。

Fractionation of nuclear and cytoplasmic ribonucleic acids from rat tissues on the methylated albumin--kieselguhr column.

作者信息

Lichtenstein A V, Shapot V S

出版信息

Biochem J. 1971 Nov;125(1):225-34. doi: 10.1042/bj1250225.

Abstract
  1. The conformation of RNA was found to affect its behaviour on methylated albumin-kieselguhr chromatography. The less regular the secondary structure of RNA, the more tightly it binds to the methylated albumin-kieselguhr column. 2. The presence of various denaturing agents (such as urea or perchlorate) in the medium while RNA was adsorbed on the column increased the resolving power of the technique as exemplified by the separation of rat liver rRNA into two distinct peaks. A special procedure for selective adsorption of the cytoplasmic DNA-like RNA on the preparative scale has been developed. Polyribosomal mRNA (rapidly labelled RNA formed in the presence of small doses of actinomycin D) can also be adsorbed selectively by the column. 3. A type of tissue specificity was detected in nuclear RNA from rat liver, kidney, thymus and spleen by using a modified salt and temperature gradient for the chromatographic fractionation (Lichtenstein, Piker & Shapot, 1967; Shapot, Lichtenstein & Piker, 1967). It was also found that cytoplasmic RNA from the different rat tissues contained no tenaciously bound fraction at all, whereas it constituted about 50% of the nuclear RNA. The problem of the possible biological function of the tenaciously bound fraction is discussed.
摘要
  1. 人们发现RNA的构象会影响其在甲基化白蛋白-硅藻土色谱上的行为。RNA二级结构越不规则,它与甲基化白蛋白-硅藻土柱的结合就越紧密。2. 当RNA吸附在柱上时,介质中存在各种变性剂(如尿素或高氯酸盐)会提高该技术的分辨能力,以大鼠肝脏rRNA分离成两个不同峰为例。已经开发出一种在制备规模上选择性吸附细胞质类DNA RNA的特殊方法。多核糖体mRNA(在小剂量放线菌素D存在下形成的快速标记RNA)也可被该柱选择性吸附。3. 通过使用改良的盐和温度梯度进行色谱分级分离,在大鼠肝脏、肾脏、胸腺和脾脏的核RNA中检测到一种组织特异性(利希滕斯坦、皮克和沙波特,1967年;沙波特、利希滕斯坦和皮克,1967年)。还发现来自不同大鼠组织的细胞质RNA根本不包含紧密结合部分,而紧密结合部分约占核RNA的50%。文中讨论了紧密结合部分可能的生物学功能问题。

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