Zhang X Y, Fittler F, Hörz W
Nucleic Acids Res. 1983 Jul 11;11(13):4287-306. doi: 10.1093/nar/11.13.4287.
The question of nucleosome phasing on African Green Monkey (AGM) alpha-satellite DNA has been addressed by employing a new approach. Nucleosome cores were prepared from AGM nuclei with micrococcal nuclease, exonuclease III and nuclease S1. The core DNA population derived from alpha-satellite DNA containing chromatin was purified from total core DNA by denaturation of the DNA, reassociation to a low Cot value, and hydroxyapatite chromatography to separate the renatured satellite fraction. After end-labeling the termini of the alpha-satellite containing core DNA fragments were mapped by high resolution gel electrophoresis relative to known restriction sites along the 172 bp repeat unit of the satellite DNA. The results show that nucleosomes occupy eight strictly defined positions on the alpha-satellite DNA which could be determined with an accuracy of +/- 1 base pair. Approximately 35% of all nucleosomes are organized in one of these frames while the other seven registers contribute about 10% each.
通过采用一种新方法,研究了非洲绿猴(AGM)α卫星DNA上核小体相位的问题。用微球菌核酸酶、核酸外切酶III和核酸酶S1从AGM细胞核中制备核小体核心。通过DNA变性、复性至低Cot值以及羟基磷灰石色谱法从总核心DNA中纯化出源自含α卫星DNA染色质的核心DNA群体,以分离复性的卫星部分。在对含α卫星的核心DNA片段的末端进行末端标记后,通过高分辨率凝胶电泳相对于卫星DNA 172 bp重复单元上的已知限制位点对其进行定位。结果表明,核小体在α卫星DNA上占据八个严格定义的位置,其精度可达±1个碱基对。所有核小体中约35%以这些框架之一排列,而其他七个排列各占约10%。