通过在碱性甲泛影酰胺密度梯度中进行等密度离心研究组蛋白在碱变性染色质中的分布。
Distribution of histones in alkali-denatured chromatin studied by isopycnic centrifugation in alkaline metrizamide density gradients.
作者信息
Russev G, Tsanev R
出版信息
Nucleic Acids Res. 1976 Mar;3(3):697-707. doi: 10.1093/nar/3.3.697.
Three types of density gradients - neutral metrizamide, alkaline NaOH-metrizamide and alkaline triethanolamine-metrizamide - were used for studying the distribution of histones between the two DNA strands in alkali-denatured chromatin. It was found possible to avoid both protein redistribution and dissociation by using triethanolamine-metrizamide density gradients at pH 10.5. Under these conditions an alkali-denatured mixture of DNA and chromatin was well separated into the original DNA and DNP. When native or sonicated chromatin was denatured at pH 12.2 and centrifuged in a triethanolamine-metrizamide density gradient at pH 10.5 no peak of free DNA appeared. These results show that both DNA strands remain associated with histone molecules upon alkaline denaturation of chromatin.
三种密度梯度——中性甲泛影酰胺、碱性氢氧化钠 - 甲泛影酰胺和碱性三乙醇胺 - 甲泛影酰胺——被用于研究碱变性染色质中组蛋白在两条DNA链之间的分布。发现在pH 10.5使用三乙醇胺 - 甲泛影酰胺密度梯度可以避免蛋白质重新分布和解离。在这些条件下，DNA和染色质的碱变性混合物能很好地分离成原始DNA和脱氧核糖核蛋白（DNP）。当天然或超声处理的染色质在pH 12.2变性并在pH 10.5的三乙醇胺 - 甲泛影酰胺密度梯度中离心时，未出现游离DNA峰。这些结果表明，在染色质碱性变性时，两条DNA链均与组蛋白分子保持结合。
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