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用于研究分离细胞核的流式细胞术方法:DNA对脱氧核糖核酸酶I的可及性及蛋白质-DNA含量

Flow cytometric methods for studying isolated nuclei: DNA accessibility to DNase I and protein-DNA content.

作者信息

Higashikubo R, Wright W D, Roti Roti J L

机构信息

Section of Cancer Biology, Washington University School of Medicine, St. Louis, Missouri 63108.

出版信息

Methods Cell Biol. 1990;33:325-36. doi: 10.1016/s0091-679x(08)60536-1.

Abstract

Two FCM methods utilizing isolated nuclei were described. A DNase I sensitivity assay, employing changes in binding and digestion kinetics of the enzyme as well as the binding of intercalating fluorochrome was used to observe structural changes of chromatin rendered by physical and chemical agents. A nuclear DNA-protein staining method was used to study changes in nuclear protein content, redistribution of populations in the cell cycle, and unbalanced growth, manifested as an extraordinary accumulation of nuclear protein brought about by physical and chemical perturbation.

摘要

描述了两种利用分离细胞核的流式细胞术方法。一种DNA酶I敏感性测定法,利用该酶结合和消化动力学的变化以及嵌入荧光染料的结合来观察物理和化学试剂引起的染色质结构变化。一种核DNA-蛋白质染色方法用于研究核蛋白含量的变化、细胞周期中群体的重新分布以及不平衡生长,不平衡生长表现为物理和化学扰动导致的核蛋白异常积累。

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