Preparation of DNA and nucleoprotein samples for AFM imaging.

Preparation of DNA and nucleoprotein samples for AFM imaging.
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DOI:
10.1016/j.micron.2010.08.011
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发表时间:
2011-02
期刊:
影响因子:
2.4
通讯作者:
Lyubchenko, Yuri L.
Lyubchenko, Yuri L.
中科院分区:
工程技术4区
文献类型:
--
作者:
Lyubchenko, Yuri L.

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对具有各种结构、拓扑结构和蛋白质复合物的 DNA 进行可靠且可重复成像的样品制备技术进行了综述。主要重点是利用云母化学功能化的方法,从而能够制备具有所需特性的表面。通过不同 DNA 结构成像的例子来说明这些方法。特别关注 AFM 在纳米尺度上对 DNA 动力学进行成像的可能性。通过将动态过程成像为局部替代结构的转变(DNA 在 H 和 B 形式之间的转变),说明了延时 AFM 在水溶液中的功能。通过一些位点特异性复合物以及染色质等系统成像的例子说明了 AFM 在蛋白质-DNA 复合物研究中的应用。回顾了蛋白质-DNA 复合物的延时 AFM 研究,包括使用高速 AFM 的最新进展。
Sample preparation techniques allowing reliable and reproducible imaging of DNA with various structures, topologies and complexes with proteins are reviewed. The major emphasis is given to methods utilizing chemical functionalization of mica, enabling preparation of the surfaces with required characteristics. The methods are illustrated by examples of imaging of different DNA structures. Special attention is given to the possibility of AFM to image the dynamics of DNA at the nanoscale. The capabilities of time-lapse AFM in aqueous solutions are illustrated by imaging of dynamic processes as transitions of local alternative structures (transition of DNA between H and B forms). The application of AFM to studies of protein-DNA complexes is illustrated by a few examples of imaging site-specific complexes, as well as such systems as chromatin. The time-lapse AFM studies of protein-DNA complexes including very recent advances with the use of high-speed AFM are reviewed.
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