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染色质核小体:分离、亚分级分离及物理特性分析

Chromatin nu bodies: isolation, subfractionation and physical characterization.

作者信息

Olins A L, Carlson R D, Wright E B, Olins D E

出版信息

Nucleic Acids Res. 1976 Dec;3(12):3271-91. doi: 10.1093/nar/3.12.3271.

Abstract

Monomer chromatin subunit particles (nu1) have been isolated in gram quantities by large-scale zonal centrifugation of micrococcal nuclease digests of chicken erythrocyte nuclei. nu1 can be stored, apparently indefinitely, frozen in 0.2 mM EDTA (pH 7.0) at less than or equal to 25 degrees C. Aliquots of the stored monomers have been subfractionated by dialysis against 0.1 M KCl buffers into a soluble fraction containing equimolar amounts of H4, H3, H2A, H2B associated with a DNA fragment of approximately 130-140 nucleotide pairs, and a precipitated fraction containing all of the histones including H5 and H1 associated with DNA fragments. The total nu1 and the KCl-soluble fraction of nu1 have been examined by sedimentation, diffusion, sedimentation equilibrium ultracentrifugation, low-angle X-ray diffraction, and electron microscopy. Physical parameters from all of these techniques are presented and correlated in this study.

摘要

通过对鸡红细胞核的微球菌核酸酶消化物进行大规模区带离心,已以克量分离出单体染色质亚基颗粒(nu1)。nu1可以在小于或等于25摄氏度的条件下,于0.2 mM EDTA(pH 7.0)中冷冻保存,显然可以无限期保存。将储存的单体等分试样通过对0.1 M KCl缓冲液进行透析,分离成一个可溶性部分,该部分含有与约130 - 140个核苷酸对的DNA片段相关的等摩尔量的H4、H3、H2A、H2B,以及一个沉淀部分,该部分含有与DNA片段相关的包括H5和H1在内的所有组蛋白。已通过沉降、扩散、沉降平衡超速离心、低角度X射线衍射和电子显微镜对总nu1和nu1的KCl可溶性部分进行了检测。本研究给出并关联了所有这些技术的物理参数。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ffc3/343173/38d45db7415a/nar00497-0031-a.jpg

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