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大鼠肝脏染色质-核膜复合体:分离与纯化

Chromatin-nuclear envelope complex from rat liver: isolation and purification.

作者信息

Prusov A N, Fais D, Polyakov V Y, Vanyushin B F

出版信息

Cell Biol Int Rep. 1980 Apr;4(4):391-8. doi: 10.1016/0309-1651(80)90220-9.

Abstract

We describe a method for the isolation of a fraction of nuclear envelope (NE) from rat liver. The method includes mild treatment of pure nuclei with either endonuclease of DNase I under low ionic strength conditions in the presence of magnesium, which allows the nucleomeric organization of the chromatin (Ch) to be preserved. The NEs were purified by centrifugation in sucrose gradients followed by floatation in sucrose. No more than 3% of the Ch present in the purified Ch-NE complexes was due to the non specific adsorption of Ch to the NE. The main components of the complex (Ch and NE) retained their in situ ultrastructure. The complex consisted of 9--10% DNA, 3--4% RNA, about 63% protein and about 24% phospholipids.

摘要

我们描述了一种从大鼠肝脏中分离核膜(NE)组分的方法。该方法包括在低离子强度条件下,于镁离子存在的情况下,用核酸内切酶或DNase I对纯细胞核进行温和处理,这样可使染色质(Ch)的核小体结构得以保留。通过在蔗糖梯度中离心,随后在蔗糖中漂浮来纯化核膜。纯化的染色质 - 核膜复合物中存在的染色质,不超过3%是由于染色质非特异性吸附到核膜上所致。复合物的主要成分(染色质和核膜)保留了它们的原位超微结构。该复合物由9 - 10%的DNA、3 - 4%的RNA、约63%的蛋白质和约24%的磷脂组成。

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