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原子力显微镜下DNA的运动及酶促降解

Motion and enzymatic degradation of DNA in the atomic force microscope.

作者信息

Bezanilla M, Drake B, Nudler E, Kashlev M, Hansma P K, Hansma H G

机构信息

Department of Physics, University of California, Santa Barbara 93106.

出版信息

Biophys J. 1994 Dec;67(6):2454-9. doi: 10.1016/S0006-3495(94)80733-7.

Abstract

The dynamics and enzymatic degradation of single DNA molecules can now be observed with the atomic force microscope. A combination of two advances has made this possible. Tapping in fluid has reduced lateral forces, which permits the imaging of loosely adsorbed molecules; and the presence of nickel ions appears to form a relatively stable bridge between the negatively charged mica and the negatively charged DNA phosphate backbone. Continuous imaging shows DNA motion and the process of DNA degradation by the nuclease DNase I. It is possible to see DNase degradation of both loosely adsorbed and tightly adsorbed DNA molecules. This method gives images in aqueous buffer of bare, uncoated DNA molecules with lengths of only a few hundred base pairs, or approximately 100 nm in length.

摘要

现在可以用原子力显微镜观察单个DNA分子的动力学和酶促降解过程。两项进展的结合使得这成为可能。在液体中轻敲减少了横向力,这使得能够对松散吸附的分子进行成像;并且镍离子的存在似乎在带负电荷的云母和带负电荷的DNA磷酸骨架之间形成了相对稳定的桥梁。连续成像显示了DNA的运动以及核酸酶DNase I对DNA的降解过程。可以看到DNase对松散吸附和紧密吸附的DNA分子的降解。这种方法可以在水性缓冲液中获得长度仅为几百个碱基对(即长度约为100纳米)的裸露、未包被的DNA分子的图像。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2503/1225630/b9c174e53d43/biophysj00068-0330-a.jpg

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