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通过电子显微镜观察动质体DNA中的弯曲螺旋。

Visualization of the bent helix in kinetoplast DNA by electron microscopy.

作者信息

Griffith J, Bleyman M, Rauch C A, Kitchin P A, Englund P T

出版信息

Cell. 1986 Aug 29;46(5):717-24. doi: 10.1016/0092-8674(86)90347-8.

Abstract

Kinetoplast DNA minicircles from the trypanosomatid Crithidia fasciculata contain a segment of approximately 200 bp which is probably more highly bent than any other DNA previously studied. Electron microscopy (EM) of relaxed minicircles (2.5 kb) revealed 200-300 bp loops within the larger circles, and the loops could also be detected on full-length linear molecules. Examination by EM of a 219 bp cloned fragment which contains the bent helix revealed that up to 70% of the molecules appeared circular whether or not the ends were cohesive. In contrast, a 207 bp fragment from pBR322 showed no circles and the fragments in general appeared much straighter than the kinetoplast fragments. Treatment of the 219 bp bent kinetoplast fragment with the drug distamycin caused a striking reduction in curvature.

摘要

来自锥虫科纤细无鞭毛体的动质体DNA微小环含有一段约200 bp的片段,该片段可能比之前研究的任何其他DNA的弯曲程度都更高。对松弛微小环(2.5 kb)进行电子显微镜(EM)观察,发现在较大的环内有200 - 300 bp的环,并且在全长线性分子上也能检测到这些环。对包含弯曲螺旋的219 bp克隆片段进行EM检查发现,无论末端是否具有粘性,高达70%的分子呈现为环状。相比之下,来自pBR322的207 bp片段未显示出环状结构,并且这些片段总体上看起来比动质体片段直得多。用药物偏端霉素处理219 bp的弯曲动质体片段会导致曲率显著降低。

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