Single filament electrophoresis of F-actin and filamentous virus fd
Single filament electrophoresis of F-actin and filamentous virus fd
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DOI：
10.1063/1.1859284
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发表时间：
2005-03-08
影响因子：
4.4
通讯作者：
Tang, JX
中科院分区：
文献类型：
--
作者：
Li, GL;Wen, Q;Tang, JX
我们开发了一种适合单分子电泳的电泳池。该装置通过直接记录荧光标记的大分子在外部电场下的运动来工作。测量了棒状多分散肌动蛋白丝（F-肌动蛋白）的电泳迁移率及其对丝的方向的依赖性。通过沿着和垂直于长丝长轴的运动之间的流体动力阻力的差异观察并定量地解释了浸渍效应。当对所有方向进行平均时，F-肌动蛋白在 50 mM KCl 和 2 mM MgCl2 中的迁移率确定为 -(8.5 +/- 0.7)x10(-5) cm(2)/(V s)。该方法还用于比较 F-肌动蛋白和 fd 病毒在混合物中的迁移率。测得 fd 病毒与 F-肌动蛋白的可靠比率为 1.26。讨论了方向相关阻力对电泳迁移率的影响，并提出了可靠测量的策略。
We have developed an electrophoretic cell suitable for single-molecule electrophoresis. The setup works for fluorescently labeled macromolecules by direct recording of their motion under an external electric field. The electrophoretic mobility of rodlike, polydisperse actin filaments (F-actin) were measured, as well as its dependence on the orientation of the filaments. A dipping effect is observed and quantitatively accounted for by the difference in hydrodynamic drag between motions along and perpendicular to the long axis of a filament. When averaged over all orientations, the mobility of F-actin in 50 mM KCl and 2 mM MgCl2 is determined to be -(8.5 +/- 0.7)x10(-5) cm(2)/(V s). This method is also used to compare the mobility of F-actin and fd virus in a mixture of them. A reliable ratio of 1.26 is measured for fd virus to F-actin. The influence of the orientation dependent drag on electrophoretic mobility is discussed and a strategy for reliable measurement is proposed.