双小核草履虫大核和小核的分离及部分特性研究
Isolation and partial characterization of macro- and micronuclei from Paramecium aurella.
作者信息
Cummings D J
出版信息
J Cell Biol. 1972 Apr;53(1):105-15. doi: 10.1083/jcb.53.1.105.
A method was developed for the isolation of macro- and micronuclei from Paramecium aurelia. This method utilized ionic and nonionic detergents to rupture the intact cells, calcium ions and spermidine were employed to protect the nuclei, and the nuclei were purified by centrifugation. Macronuclei consisted of 22% DNA, 10% RNA, and 68% protein. Micronuclei were composed of 9% DNA, 11% RNA, and 80% protein. DNA from both macro- and micronuclei had a density of 1.687 g/cc in CsCl and 1.417 g/cc in Cs(2)SO(4). These values corresponded to G + C content of about 23%. The RNA of macronuclei was examined by gel electrophoresis, and two high molecular weight species were identified having molecular Weights of 1.3 x 10(6) and 2.8 x 10(6) daltons. Three syngens were studied, and in each case the conditions for isolation of the nuclei were the same and no differences were observed in the properties of the nuclei.
开发了一种从双小核草履虫中分离大核和小核的方法。该方法利用离子和非离子去污剂使完整细胞破裂，使用钙离子和亚精胺保护细胞核，并通过离心纯化细胞核。大核由22%的DNA、10%的RNA和68%的蛋白质组成。小核由9%的DNA、11%的RNA和80%的蛋白质组成。大核和小核的DNA在CsCl中的密度为1.687 g/cc，在Cs₂SO₄中的密度为1.417 g/cc。这些值对应于约23%的G + C含量。通过凝胶电泳检查大核的RNA，鉴定出两种高分子量物质，分子量分别为1.3×10⁶和2.8×10⁶道尔顿。研究了三个同宗配合物，在每种情况下，细胞核的分离条件相同，并且在细胞核的性质上未观察到差异。
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