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大肠杆菌RNA聚合酶在金上的电沉积过程以及大肠杆菌RNA聚合酶在云母上的沉积用于扫描力显微镜观察。

Electrodeposition procedure of E. coli RNA polymerase onto gold and deposition of E. coli RNA polymerase onto mica for observation with scanning force microscopy.

作者信息

Keller R W, Keller D J, Bear D, Vasenka J, Bustamante C

机构信息

Department of Chemistry, University of New Mexico, Albuquerque 87131.

出版信息

Ultramicroscopy. 1992 Jul;42-44 ( Pt B):1173-80. doi: 10.1016/0304-3991(92)90420-o.

Abstract

Molecules of the transcriptional enzyme E. coli RNA polymerase (RNAP) have been deposited using three different deposition methods: (1) passive adsorption onto gold, (2) electrochemical adsorption onto gold and (3) adsorption onto mica. In all cases SFM imaging was straightforward and reliable, and surface coverage by the protein varied with deposition conditions as expected. To determine the nature of the electrochemical treatment on the gold substrate, cyclic voltammetry was performed with various chemical solutions. Finally, a comparison is made between the SFM images of RNAP obtained with these methods and STM images obtained earlier. Both STM and SFM show strikingly similar results; however, heights and widths of individual molecules differ.

摘要

已使用三种不同的沉积方法来放置转录酶大肠杆菌RNA聚合酶(RNAP)的分子:(1)被动吸附到金上,(2)电化学吸附到金上,以及(3)吸附到云母上。在所有情况下,扫描力显微镜(SFM)成像都直接且可靠,并且蛋白质的表面覆盖率随沉积条件而变化,正如预期的那样。为了确定金基底上电化学处理的性质,用各种化学溶液进行了循环伏安法测定。最后,对用这些方法获得的RNAP的SFM图像与早期获得的扫描隧道显微镜(STM)图像进行了比较。STM和SFM都显示出惊人的相似结果;然而,单个分子的高度和宽度有所不同。

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