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由分子拥挤剂聚乙二醇凝聚的DNA的荧光动力学

Fluorescence dynamics of DNA condensed by the molecular crowding agent poly(ethylene glycol).

作者信息

Kombrabail Mamata H, Krishnamoorthy G

机构信息

Department of Chemical Sciences, Tata Institute of Fundamental Research, Mumbai, India.

出版信息

J Fluoresc. 2005 Sep;15(5):741-7. doi: 10.1007/s10895-005-2982-8.

Abstract

Condensation of extended DNA into compact structures is encountered in a variety of situations, both natural and artificial. While condensation of DNA has been routinely carried out by the use of multivalent cations, cationic lipids, detergents, and polyvalent cationic polymers, the use of molecular crowding agents in condensing DNA is rather striking. In this work, we have studied the dynamics of plasmid DNA condensed in the presence of a molecular crowding agent, polyethylene glycol (PEG). Steady-state and time-resolved fluorescence of the recently established condensation-indicating DNA binder, YOYO-1 [G. Krishnamoorthy, G. Duportail, and Y. Mely (2002), Biochemistry 41, 15277-15287] was used in inferring the dynamic aspects of DNA condensates. It is shown that DNA condensed by PEG is more flexible and less compact when compared to DNA condensed by binding agents such as polyethyleneimine. The relevance of such differences in dynamics toward functional aspects of condensed DNA is discussed.

摘要

在各种自然和人工情况下,都会遇到将伸展的DNA浓缩成紧密结构的情况。虽然DNA的浓缩通常是通过使用多价阳离子、阳离子脂质、去污剂和多价阳离子聚合物来实现的,但使用分子拥挤剂来浓缩DNA却相当引人注目。在这项工作中,我们研究了在分子拥挤剂聚乙二醇(PEG)存在下浓缩的质粒DNA的动力学。最近建立的用于指示DNA浓缩的结合剂YOYO-1 [G. Krishnamoorthy, G. Duportail, and Y. Mely (2002), Biochemistry 41, 15277-15287] 的稳态和时间分辨荧光被用于推断DNA凝聚物的动态方面。结果表明,与通过诸如聚乙烯亚胺等结合剂浓缩的DNA相比,PEG浓缩的DNA更具柔韧性且结构更不紧密。讨论了这种动力学差异对浓缩DNA功能方面的相关性。

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