Seligy V L, Poon N H
Nucleic Acids Res. 1978 Jul;5(7):2233-52. doi: 10.1093/nar/5.7.2233.
Mononucleosomes prepared from goose erythrocyte nuclei exhibited limited heterogeneity with respect to number of electrophoretic components, histones and DNA composition. The components differ slightly in ionic strength induced self-association. Thermal denaturation of each component gave only two dominant, highly cooperative, melting transitions, T" and T"'. Urea and trypsin were used to establish the differential lability of these two transitions. Comparison of the morphologies of the mononucleosomes at various stages throughout the melting profile indicated that the 13.3 +/- 1.5 nm diameter mononucleosomes start to disrupt only in the latter half of transition T" and do not unfold until after reaching T"'. The resultant, open ended (17.4 +/- 2.2 nm diameter) toroids are still largely negatively staining and much more uniform in shape if fixed simultaneously with gluteraldehyde.
从鹅红细胞核制备的单核小体在电泳成分数量、组蛋白和DNA组成方面表现出有限的异质性。这些成分在离子强度诱导的自缔合方面略有不同。每个成分的热变性仅产生两个主要的、高度协同的解链转变,即T" 和T"'。使用尿素和胰蛋白酶来确定这两个转变的不同稳定性。在整个解链过程的不同阶段对单核小体形态的比较表明,直径为13.3±1.5 nm的单核小体仅在转变T" 的后半段开始破坏,直到达到T"' 后才展开。如果与戊二醛同时固定,所得的开口环形结构(直径为17.4±2.2 nm)在很大程度上仍为负染色,且形状更加均匀。