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利用荧光探针[Ru(phen)₂dppz]²⁺对DNA与聚酰胺-胺型树枝状大分子(PAMAM)之间相互作用的研究

Studies on the interactions between DNA and PAMAM with fluorescent probe [Ru(phen)2 dppz]2+.

作者信息

Zhou Linzhu, Gan Lu, Li Haibing, Yang Xiangliang

机构信息

College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.

出版信息

J Pharm Biomed Anal. 2007 Jan 4;43(1):330-4. doi: 10.1016/j.jpba.2006.06.021. Epub 2006 Jul 26.

Abstract

The interactions between plasmid DNA and cationic polymers are of interest for their potential biological applications. In this paper, the interactions of DNA with polyamidoamine (PAMAM) dendrimer were studied by fluorescence spectroscopy and stopped-flow technique. A rapid and reproducible fluorescent assay method had been developed for assessing PAMAM and DNA interactions using [Ru(phen)2 dppz]2+ as a probe. We further studied the kinetics of PAMAM binding to DNA and the reverse process of DNA dissociation from the complexes. The results indicated that DNA condensation was the rate-determining step during the complexation process, while DNA unfolding and expansion was the rate-determining step during the DNA dissociation process. At N/P ratios before reaching the thermodynamically most stable state, the complexes of DNA and PAMAM were incompact and could dissociate to some extent. And at N/P ratios above 2.0, DNA was fully condensed by PAMAM and dissociation was increasingly difficult. These results provided some useful instructions for self-assembling and disassembling of DNA as well as efficient gene delivery applications.

摘要

质粒DNA与阳离子聚合物之间的相互作用因其潜在的生物学应用而备受关注。本文采用荧光光谱法和停流技术研究了DNA与聚酰胺-胺(PAMAM)树枝状大分子的相互作用。以[Ru(phen)2 dppz]2+为探针,建立了一种快速、可重复的荧光检测方法,用于评估PAMAM与DNA的相互作用。我们进一步研究了PAMAM与DNA结合的动力学以及DNA从复合物中解离的逆过程。结果表明,DNA凝聚是复合过程中的速率决定步骤,而DNA解折叠和伸展是DNA解离过程中的速率决定步骤。在达到热力学最稳定状态之前的N/P比下,DNA与PAMAM的复合物不紧密,在一定程度上可以解离。而在N/P比高于2.0时,DNA被PAMAM完全凝聚,解离变得越来越困难。这些结果为DNA的自组装和解组装以及高效基因传递应用提供了一些有用的指导。

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