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在模型薄膜中研究染色质紧密程度对福尔根-席夫反应化学计量的影响。I. 含分离脱氧核糖核酸薄膜的理论动力学与实验

The influence of chromatin compactness on the stoichiometry of the Feulgen-Schiff procedure studied in model films. I. Theoretical kinetics and experiments with films containing isolated deoxyribonucleic acid.

作者信息

Duijndam W A, van Duijn P

出版信息

J Histochem Cytochem. 1975 Dec;23(12):882-90. doi: 10.1177/23.12.53248.

Abstract

Theoretical considerations on the expected kinetics of the course of the Feulgen-Schiff reaction show that the leveling off of the first part of the Feulgen hydrolysis curve can be explained by the gradual conversion of deoxyribonucleic acid (DNA) to apurinic acid (APA). In addition, depolymerization of DNA caused by the acid used for hydrolysis can account for the decline after a maximum is reached in this curve. With the aid of polyacrylamide model films containing DNA, a detailed study was made both of the process of purine liberation which results in the formation of APA and of the depolymerization processes which cause losses of stainable material. The liberation of purine bases was analyzed by ultraviolet absorbance measurements and by gel chromatography of the neutralized hydrolysing acid. APA concentration was monitored by following the loss of ultraviolet absorbance associated with the purine losses. The depolymerization process was followed by phosphorus determinations. The experimental results were found to be in accordance with the kinetics expected from the theoretical model.

摘要

对福尔根-席夫反应进程预期动力学的理论考量表明,福尔根水解曲线第一部分的趋于平稳可通过脱氧核糖核酸(DNA)逐渐转化为脱嘌呤酸(APA)来解释。此外,水解所用酸导致的DNA解聚可解释该曲线达到最大值后出现的下降。借助含有DNA的聚丙烯酰胺模型膜,对导致APA形成的嘌呤释放过程以及引起可染色物质损失的解聚过程进行了详细研究。通过紫外吸光度测量和中和水解酸的凝胶色谱分析嘌呤碱的释放。通过跟踪与嘌呤损失相关的紫外吸光度损失来监测APA浓度。通过磷测定来跟踪解聚过程。实验结果与理论模型预期的动力学一致。

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