Wittig B, Wittig S
Nucleic Acids Res. 1977 Nov;4(11):3901-17. doi: 10.1093/nar/4.11.3901.
The fractionation of gram quantities of nuclease digested chromatin from chicken embryos into nucleosome mono-, di-, tri-, and tetramers is described in detail. Each of these nucleosomal species contains a fraction soluble in 0-1 M KC1 that decreases with increasing repeat number. Less histone H1 is associated with the nucleosome fractions soluble as compared to the respective fractions precipitated in 0.1 M KC1. Thermal denaturation profiles of the four nucleosomal species are monophasic. The same Tm of 78 degrees C has been determined for the KC1-soluble nucleosomes and for the KC1-insoluble monomer. The Tm of the KC1-insoluble oligomers is 79.8 degrees C. Multiphasic melting curves were recorded for nucleosomal material that was concentrated by lyophilisation or stored at 4 degrees C in 0.25 mM EDTA. Total nucleosome mono-, di-, tri-, and tetramers (consisting of both the fraction soluble and insoluble in 0.1 M KC1) have been analyzed concerning their sedimentation, diffusion, partial specific volume, and molecular weight and compared with the sedimentation and molecular weight data of KC1-soluble nucleosome mono- and tetramers.
详细描述了从鸡胚中对克数量的核酸酶消化染色质进行分级分离,得到核小体单体、二聚体、三聚体和四聚体的过程。这些核小体种类中的每一种都含有一部分可溶于0 - 1M KCl的成分,该成分会随着重复数目的增加而减少。与在0.1M KCl中沉淀的相应部分相比,与可溶的核小体部分结合的组蛋白H1较少。这四种核小体种类的热变性曲线是单相的。已确定KCl可溶核小体和KCl不溶单体的相同熔点为78℃。KCl不溶寡聚体的熔点为79.8℃。对于通过冻干浓缩或在0.25mM EDTA中于4℃储存的核小体物质,记录到多相熔解曲线。已对总的核小体单体、二聚体、三聚体和四聚体(包括在0.1M KCl中可溶和不可溶的部分)的沉降、扩散、比容和分子量进行了分析,并与KCl可溶核小体单体和四聚体的沉降和分子量数据进行了比较。