Cytofluorometric studies on conformation of nucleic acids in situ. II. Denaturation of deoxyribonucleic acid.
Cytofluorometric studies on conformation of nucleic acids in situ. II. Denaturation of deoxyribonucleic acid.
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原位核酸构象的细胞荧光研究。
DOI：
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发表时间：
1976
影响因子：
3.2
通讯作者：
M. Melamed
中科院分区：
文献类型：
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作者：
Z. Darżynkiewicz;F. Traganos;Z. Arlin;A. T. Sharpless;M. Melamed
用细胞荧光法研究了单个完整细胞中脱氧核糖核酸（DNA）的原位热变性。这种方法使我们能够研究DNA变性的二价阳离子的存在下，在浓度报告是必要的，以保持天然结构的核染色质。在这些条件下，DNA变性的模式与在乙二胺四乙酸盐或柠檬酸盐存在下研究时非常不同。结果表明，与二价阳离子的存在，组蛋白的基本电荷分布更均匀沿着整个核DNA。当在1 mM MgCl2存在下测定时，各种细胞类型对DNA变性的敏感性表现出很大差异。人类淋巴细胞、单核细胞和某些种类的人类白血病细胞显示出足够大的差异，可以用作在混合样品中识别它们的参数。该方法在染色质构象的基础研究中可能的应用，并作为诊断细胞学或人类白血病分类中的细胞识别工具。
Thermal denaturation of deoxyribonucleic acid (DNA) in situ in individual unbroken cells is studied by a cytofluorometric method. This method allows us to investigate DNA denaturation in the presence of divalent cations at concentrations reported to be necessary to maintain native structure of nuclear chromatin. Under these conditions the pattern of DNA denaturation is very different than when studied in the presence of ethylenediaminetetraacetate or citrate. The results suggest that with divalent cations present, the histone basic charges are more uniformly distributed along whole nuclear DNA. Various cell types exhibit great differences in sensitivity to DNA denaturation when assayed in the presence of 1 mM MgCl2. Human lymphocytes, monocytes and certain kinds of human leukemic cells show differences large enough to be used as a parameter for their recognition in mixed samples. Possible applications of the method in basic research on chromatin conformation and as a tool for cell recognition in diagnostic cytology or in the classification of human leukemia are proposed.