Szent-Gyorgyi C, Isenberg I
Nucleic Acids Res. 1983 Jun 11;11(11):3717-36. doi: 10.1093/nar/11.11.3717.
We have developed a method of preparing yeast chromatin that facilitates the analysis of nucleoprotein organization. Yeast chromatin, isolated as an insoluble complex, is digested with micrococcal nuclease and fractionated into major insoluble and soluble fractions. No nucleosomal repeat is seen early in digestion for the insoluble fraction. Nucleosomal complexes of the soluble fraction are excised by nuclease in a distinctive non-random pattern; they are markedly depleted in mononucleosomes. When we analyze the soluble material by high resolution native electrophoresis, we find that the nucleoproteins resolve into two bands for each DNA multimer of the nucleosomal repeat. Our results suggest that there are structural similarities between bulk yeast chromatin and chromatin configurations found in transcribing genes of complex eukaryotes.
我们开发了一种制备酵母染色质的方法，该方法有助于分析核蛋白的组织。作为不溶性复合物分离出的酵母染色质，用微球菌核酸酶消化并分级分离成主要的不溶性和可溶性部分。在消化早期，不溶性部分未观察到核小体重复序列。可溶性部分的核小体复合物被核酸酶以独特的非随机模式切除；它们在单核小体中明显减少。当我们通过高分辨率天然电泳分析可溶性物质时，我们发现核蛋白对于核小体重复序列的每个DNA多聚体都分解成两条带。我们的结果表明，大量酵母染色质与复杂真核生物转录基因中发现的染色质构型之间存在结构相似性。
