McGhee J D, Kimmel C B
Chromosoma. 1975 Sep 26;52(2):189-205. doi: 10.1007/BF00326267.
If micrococcal nuclease is allowed to digest chromatin as it exists inside intact nuclei isolated from mouse myeloma tissue culture cells, more than 60% of the DNA can be isolated as a homogeneous fragment on a sucrose gradient. Analytical ultracentrifugation indicates that the protected DNA is native, unnicked, and about 140 +/- 10 base pairs long. After less extensive nuclease digestion, the protected DNA migrates in gels in lengths which are integral multiples of this 140 base pair "monomer" band. A submonomer band, 105 "/- 10 base pairs long, can also be detected. Similar digestion patterns were obtained by two different nuclear isolation procedures and even when intact cells were gently lysed directly in the digestion medium. These results confirm and extend the chromatin digestion studies of previous investigators and provide support for a subunit model for eukaryotic chromatin. The single strand specific S1 nuclease did not digest intranuclear chromatin under the conditions used.
如果用微球菌核酸酶消化从小鼠骨髓瘤组织培养细胞中分离出的完整细胞核内的染色质,超过60%的DNA可在蔗糖梯度上作为均匀片段分离出来。分析超速离心表明,受保护的DNA是天然的、未切口的,长度约为140±10个碱基对。核酸酶消化程度较低时,受保护的DNA在凝胶中迁移的长度是这个140碱基对“单体”条带的整数倍。还能检测到一条长度为105±10个碱基对的亚单体条带。通过两种不同的细胞核分离方法,甚至当完整细胞直接在消化培养基中轻轻裂解时,都获得了类似的消化模式。这些结果证实并扩展了先前研究者的染色质消化研究,并为真核染色质的亚基模型提供了支持。在所使用的条件下,单链特异性S1核酸酶不会消化核内染色质。