Isolation of nucleoli in a medium containing spermine and magnesium acetate.
Isolation of nucleoli in a medium containing spermine and magnesium acetate.
复制标题
在含有精胺和醋酸镁的培养基中分离核仁。
DOI：
10.1016/0014-4827(67)90235-2
复制
发表时间：
1967
影响因子：
3.7
通讯作者：
J. Hamilton
中科院分区：
文献类型：
--
作者：
H. Busch;K. Narayan;J. Hamilton
研究了聚阳离子对核仁超微结构的影响。当肿瘤细胞在缺乏二价离子、多胺或组蛋白的0.25M蔗糖中均质化时，核仁结构难以识别并且核仁成分似乎是分散的。在用于均质化的蔗糖培养基中添加富含精氨酸的组蛋白和一些稍微富含赖氨酸的组蛋白以及多胺，在超微结构形态方面保留了核仁结构。将鱼精蛋白、精胺和亚精胺添加到含有蔗糖的匀浆介质中，产生致密的核仁。这些结果表明，可以开发一种分离核仁的实用方法，其中使用多胺，并且发现15 mMMg2+和0.25 mM精胺的组合可以为0.25M蔗糖提供令人满意的添加，以维持核仁完整性。发现从在含有该组合的培养基中匀浆的肝细胞中分离的核仁在形态和组成上与用含有钙离子的蔗糖培养基获得的核仁相似。产量基本相同。
Studies are presented on the effects of polycations on the ultrastructure of nucleoli. When tumor cells were homogenized in 0.25Msucrose lacking divalent ions, polyamines or histones, the nucleolar structure was difficult to identify and nucleolar components appeared to be dispersed. Addition of arginine-rich histones and some slightly lysine-rich histones as well as polyamines to the sucrose medium used for homogenization preserved nucleolar structure with respect to ultrastructural morphology. Addition of protamines, spermine and spermidine to the homogenization medium containing sucrose produced compact and dense nucleoli. These results suggested that a practical procedure for isolation of nucleoli could be developed in which the polyamines were used and a combination of 15 mMMg2+and 0.25 mMspermine was found to provide a satisfactory addition to 0.25Msucrose for maintenance of nucleolar integrity. Nucleoli isolated from liver cells homogenized in a medium containing this combination were found to be similar in morphology and composition to those obtained with the sucrose medium containing calcium ion. The yields were essentially the same.