Henney H R, Yee M
Cytobios. 1979;24(94):103-16.
A method was established for the isolation and purification of nuclei in high yield from the microplasmodia of Physarum flavicomum. Purified nuclei was resistant to breakage by methods commonly employed for isolated plant and animal nuclei. Incubation of nuclei with 5 mM dithiothreitol at pH 9.2 was found to be the simplest and most effective method for breaking the nuclei. Several methods for the extraction of nuclear protein were compared. Incubation of nuclear lysates with either 2 M NaCl, with or without 5 M urea, or 1 M CaCl2 resulted in the extraction of nuclear actin together with histones. The histones were chemically fractionated into the five basic groups common to other eucaryotic tissue. Amino acid analyses of the total histone were also performed. Nuclear actin was found to have a molecular weight of 41,000 +/- 4,000 daltons as determined by SDS polyacrylamide gel electrophoresis. The amino acid composition of the nuclear actin was established.
建立了一种从黄绒绒泡菌的微原质团中高产分离和纯化细胞核的方法。纯化后的细胞核对通常用于分离植物和动物细胞核的方法具有抗破碎性。发现在pH 9.2条件下用5 mM二硫苏糖醇孵育细胞核是破碎细胞核最简单且最有效的方法。比较了几种提取核蛋白的方法。用2 M NaCl（有无5 M尿素）或1 M CaCl₂孵育核裂解物会导致核肌动蛋白与组蛋白一起被提取出来。组蛋白被化学分离为其他真核组织共有的五个基本组。还对总组蛋白进行了氨基酸分析。通过SDS聚丙烯酰胺凝胶电泳测定，发现核肌动蛋白的分子量为41,000±4,000道尔顿。确定了核肌动蛋白的氨基酸组成。
