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Stretching of single collapsed DNA molecules.

机译:拉伸单个折叠的DNA分子。

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摘要

The elastic response of single plasmid and lambda phage DNA molecules was probed using optical tweezers at concentrations of trivalent cations that provoked DNA condensation in bulk. For uncondensed plasmids, the persistence length, P, decreased with increasing spermidine concentration before reaching a limiting value 40 nm. When condensed plasmids were stretched, two types of behavior were observed: a stick-release pattern and a plateau at approximately 20 pN. These behaviors are attributed to unpacking from a condensed structure, such as coiled DNA. Similarly, condensing concentrations of hexaammine cobalt(III) (CoHex) and spermidine induced extensive changes in the low and high force elasticity of lambda DNA. The high force (5-15 pN) entropic elasticity showed worm-like chain (WLC) behavior, with P two- to fivefold lower than in low monovalent salt. At lower forces, a 14-pN plateau abruptly appeared. This corresponds to an intramolecular attraction of 0.083-0.33 kT/bp, consistent with osmotic stress measurements in bulk condensed DNA. The intramolecular attractive force with CoHex is larger than with spermidine, consistent with the greater efficiency with which CoHex condenses DNA in bulk. The transition from WLC behavior to condensation occurs at an extension about 85% of the contour length, permitting looping and nucleation of condensation. Approximately half as many base pairs are required to nucleate collapse in a stretched chain when CoHex is the condensing agent.
机译:使用光学镊子在三价阳离子浓度下探测单个质粒和λ噬菌体DNA分子的弹性响应,从而引起大量DNA缩合。对于未缩合的质粒,在达到40 nm极限值之前,持久性长度P随着亚精胺浓度的增加而降低。拉伸浓缩的质粒时,观察到两种类型的行为:粘着释放模式和大约20 pN的平稳期。这些行为归因于从压缩结构(例如卷曲的DNA)中解包。同样,六氨合钴(III)(CoHex)和亚精胺的浓缩浓度引起了λDNA的低和高力弹性的广泛变化。高力(5-15 pN)的熵弹性表现出蠕虫状链(WLC)行为,其中P比低一价盐低2至5倍。在较低的力量下,突然出现14 p​​N的平台。这对应于0.083-0.33 kT / bp的分子内吸引力,与大体积浓缩DNA中的渗透压测量一致。 CoHex的分子内吸引力大于亚精胺,这与CoHex大量凝聚DNA的更高效率相吻合。从WLC行为到凝结的过渡发生在轮廓长度的大约85%的延伸处,允许凝结成环和成核。当CoHex为缩合剂时,要使成核的链在拉伸链中塌陷,需要大约一半的碱基对。

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