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氯化钠对福尔根酸水解过程中DNA片段提取的影响。

The effect of sodium chloride on the extraction of DNA fragments during Feulgen acid hydrolysis.

作者信息

Kjellstrand P T

出版信息

Histochem J. 1977 May;9(3):357-67. doi: 10.1007/BF01004771.

Abstract

Feulgen acid hydrolysis was performed on ascites tumour cells labelled with radioactive DNA-precursors. The development of fragments of apurinic acid and the extraction of purines were studied by monitoring the variations in the extraction rate during the hydrolysis when sodium chloride was either present or absent from the hydrolysis solution. The changes in the rate of extraction of purines and the alterations in the initial retardation of the apurinic acid extracting process followed approximately the same pattern. The extractability of apurinic acid fragments during hydrolysis in 0.3 M HCl was found to be a maximum when the sodium chloride concentration was about 1 M. Sudden exchange experiments, in which acid was substituted for sodium chloride after various times of hydrolysis, revealed a successive shortening of the extractable fragments during the low acid concentration hydrolysis. The results strengthen the view that, during hydrolysis, apurinic acid is lost from the cells through a reaction whose form is determined, first, by an initial retardation of the depolymerization, second, by the maximum length at which fragments developed through the depolymerization become soluble and are lost by diffusion, and last, at low acid concentrations, by a mechanism whose influence is equivalent to the presence of bonds between the fragments and an unextractable stable structure.

摘要

对用放射性DNA前体标记的腹水肿瘤细胞进行福尔根酸水解。通过监测水解过程中水解溶液中存在或不存在氯化钠时提取率的变化,研究了脱嘌呤酸片段的形成和嘌呤的提取。嘌呤提取率的变化和脱嘌呤酸提取过程初始延迟的改变大致遵循相同的模式。发现在0.3 M HCl中水解时,当氯化钠浓度约为1 M时,脱嘌呤酸片段的可提取性最大。在不同水解时间后用酸代替氯化钠的突然交换实验表明,在低酸浓度水解过程中,可提取片段连续缩短。结果强化了这样一种观点,即在水解过程中,脱嘌呤酸通过一种反应从细胞中丢失,该反应的形式首先由解聚的初始延迟决定,其次由通过解聚形成的片段变得可溶并通过扩散丢失的最大长度决定,最后在低酸浓度下,由一种其影响等同于片段与不可提取的稳定结构之间存在键的机制决定。

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