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悬滴法对670千碱基对DNA进行视频光学显微镜观察:DNA包膜的形状

Video light microscopy of 670-kb DNA in a hanging drop: shape of the envelope of DNA.

作者信息

Serwer P, Estrada A, Harris R A

机构信息

Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760, USA.

出版信息

Biophys J. 1995 Dec;69(6):2649-60. doi: 10.1016/S0006-3495(95)80135-9.

Abstract

Although its conformation has not been observed directly, double-stranded DNA in solution is usually assumed to be randomly coiled at the level of the DNA double helix. By video light microscopy of ethidium-stained DNA at equilibrium in a nonturbulent hanging drop, in the present study, the 670 kb linear bacteriophage G DNA is found to form a flexible filament that has on average 17 double helical segments across its width. This flexible filament 1) has both asymmetry and dimensions expected of a random coil and 2) has ends that move according to the statistics expected of a random walk. After unraveling the flexible filament-associated DNA double helix near the surface of a hanging drop, recompaction occurs without perceptible rotation of the DNA. Both conformational change and intermolecular tangling of the DNA are observed when G DNA undergoes nondiffusive motion in a hanging drop. The characteristics of the G DNA flexible filament are explained by the assumption that the flexible filament is a random coil of double helical segments that are unperturbed by motion of the suspending medium.

摘要

尽管尚未直接观察到双链DNA的构象,但通常认为溶液中的双链DNA在DNA双螺旋水平上是随机卷曲的。在本研究中,通过对在非湍流悬滴中处于平衡状态的溴化乙锭染色DNA进行视频光学显微镜观察,发现670 kb的线性噬菌体G DNA形成了一条柔性细丝,其宽度平均有17个双螺旋片段。这条柔性细丝1)具有随机卷曲所预期的不对称性和尺寸,2)其末端的移动符合随机游走的统计规律。在悬滴表面附近解开与柔性细丝相关的DNA双螺旋后,重新压缩发生时DNA没有明显的旋转。当噬菌体G DNA在悬滴中进行非扩散运动时,会观察到DNA的构象变化和分子间缠结。假设柔性细丝是由双螺旋片段组成的随机卷曲,且不受悬浮介质运动的干扰,这可以解释噬菌体G DNA柔性细丝的特征。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/598f/1236502/e46fda7c13ad/biophysj00054-0471-a.jpg

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