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通过去除抗衡离子稳定 DNA 聚集。

Stable DNA aggregation by removal of counterions.

机构信息

Department of Chemistry and Centre for Research on Biomolecular Interactions, York University , Toronto, Ontario M3J 1P3, Canada.

出版信息

Anal Chem. 2013 Nov 5;85(21):10004-7. doi: 10.1021/ac402645n. Epub 2013 Oct 8.

Abstract

Negatively charged DNA can form extremely stable complexes with positively charged ions. These counterions are very difficult to remove from DNA; therefore, little is known about DNA behavior in their deficiency. We investigated whether removal of counterions from the strongly bound counterion layer would elicit any novel DNA properties or behaviors. In order to remove the tightly bound counterions, we used dialysis against deionized water in the presence of a strong (0.6 kV/cm) electric field. The electric field promoted the dissociation of the DNA-counterion complexes, while dialysis facilitated irreversible partitioning of counterions and DNA. Counterintuitively, when deprived of counterions, DNA precipitated from the solution into amorphous aggregates. The aggregates remained stable even when the electric field was turned off but readily redissolved when counterions were reintroduced. The phenomenon is likely explained by attraction of like-charged DNA polyions due to entropic-stabilization of condensed counterion layers.

摘要

带负电荷的 DNA 可以与带正电荷的离子形成极其稳定的复合物。这些反离子很难从 DNA 中去除;因此,人们对 DNA 在缺乏反离子时的行为知之甚少。我们研究了从强结合反离子层中去除反离子是否会引发任何新的 DNA 特性或行为。为了去除紧密结合的反离子,我们在存在强电场(0.6 kV/cm)的情况下用去离子水透析。电场促进了 DNA-反离子复合物的解离,而透析促进了反离子和 DNA 的不可逆分配。出人意料的是,当 DNA 失去反离子时,它会从溶液中沉淀出来形成无定形聚集体。这些聚集体即使在电场关闭时也保持稳定,但当重新引入反离子时很容易重新溶解。这种现象可能是由于凝聚的反离子层的熵稳定化导致相同电荷的 DNA 聚离子的吸引力所致。

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