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脉冲场琼脂糖凝胶电泳在形态发生研究中的应用:双链DNA在噬菌体衣壳中的包装

Pulsed field agarose gel electrophoresis in the study of morphogenesis: packaging of double-stranded DNA in the capsids of bacteriophages.

作者信息

Serwer P, Hayes S J, Moreno E T, Louie D, Watson R H, Son M

机构信息

Department of Biochemistry, University of Texas Health Science Center, San Antonio 78284-7760.

出版信息

Electrophoresis. 1993 Apr;14(4):271-7. doi: 10.1002/elps.1150140148.

Abstract

To understand how comparatively simple macromolecular components become biological systems, studies are made of the morphogenesis of bacteriophages. Pulsed field agarose gel electrophoresis (PFGE) has contributed to these studies by: (i) improving the length resolution of both mature, linear, double-stranded bacteriophage DNAs and the concatemers formed both in vivo and in vitro by the end-to-end joining of these mature bacteriophage DNAs, (ii) improving the resolution of circular conformers of bacteriophage DNAs, (iii) improving the resolution of linear single-stranded bacteriophage DNAs, (iv) providing a comparatively simple technique for analyzing protein-DNA complexes, and (v) providing a solid-phase quantitative assay for all forms of bacteriophage DNA; solid-phase assays are both less complex and more efficient than liquid-phase assays such as rate zonal centrifugation. Conversely, studies of bacteriophages have contributed to PFGE the DNA standards used for determining the length of nonbacteriophage DNAs. Among the solid-phase assays based on PFGE is an assay for excluded volume effects.

摘要

为了理解相对简单的大分子组分如何形成生物系统,人们对噬菌体的形态发生进行了研究。脉冲场琼脂糖凝胶电泳(PFGE)通过以下方式对这些研究做出了贡献:(i)提高成熟线性双链噬菌体DNA以及这些成熟噬菌体DNA通过端对端连接在体内和体外形成的多联体的长度分辨率;(ii)提高噬菌体DNA环状构象体的分辨率;(iii)提高线性单链噬菌体DNA的分辨率;(iv)提供一种相对简单的分析蛋白质-DNA复合物的技术;(v)为所有形式的噬菌体DNA提供一种固相定量测定法;固相测定法比速率区带离心等液相测定法更简单且更高效。相反,对噬菌体的研究也为PFGE贡献了用于确定非噬菌体DNA长度的DNA标准。基于PFGE的固相测定法之一是排除体积效应测定法。

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