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通过冷冻电子显微镜测定DNA持久长度。区分静态和动态因素对DNA表观持久长度的贡献。

Determination of DNA persistence length by cryo-electron microscopy. Separation of the static and dynamic contributions to the apparent persistence length of DNA.

作者信息

Bednar J, Furrer P, Katritch V, Stasiak A Z, Dubochet J, Stasiak A

机构信息

Laboratoire d'Analyse Ultrastructurale, Université de Lausanne, Lausanne-Dorigny, Switzerland.

出版信息

J Mol Biol. 1995 Dec 8;254(4):579-94. doi: 10.1006/jmbi.1995.0640.

Abstract

Axial deflection of DNA molecules in solution results from thermal motion and intrinsic curvature related to the DNA sequence. In order to measure directly the contribution of thermal motion we constructed intrinsically straight DNA molecules and measured their persistence length by cryo-electron microscopy. The persistence length of such intrinsically straight DNA molecules suspended in thin layers of cryo-vitrified solutions is about 80 nm. In order to test our experimental approach, we measured the apparent persistence length of DNA molecules with natural "random" sequences. The result of about 45 nm is consistent with the generally accepted value of the apparent persistence length of natural DNA sequences. By comparing the apparent persistence length to intrinsically straight DNA with that of natural DNA, it is possible to determine both the dynamic and the static contributions to the apparent persistence length.

摘要

溶液中DNA分子的轴向偏转源于热运动以及与DNA序列相关的固有曲率。为了直接测量热运动的贡献,我们构建了固有直链DNA分子,并通过冷冻电子显微镜测量其持久长度。悬浮在冷冻玻璃化溶液薄层中的此类固有直链DNA分子的持久长度约为80纳米。为了测试我们的实验方法,我们测量了具有天然“随机”序列的DNA分子的表观持久长度。约45纳米的结果与天然DNA序列表观持久长度的普遍接受值一致。通过比较天然DNA与固有直链DNA的表观持久长度,可以确定对表观持久长度的动态和静态贡献。

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