凝胶中DNA的检测：应用脉冲场琼脂糖凝胶电泳研究病毒DNA代谢

In-gel detection of DNA : application to study of viral DNA metabolism by use of pulsed-field agarose gel electrophoresis.

作者信息

Sewer P, Watson R H, Son M

机构信息

Department of Biochemistry, The University of Texas Health Science Center, San Antonio, TX.

出版信息

Methods Mol Biol. 1992;12:333-43. doi: 10.1385/0-89603-229-9:333.

DOI:10.1385/0-89603-229-9:333

PMID:21409643

Abstract

The introduction of pulsed-field agarose gel electrophoresis (PFGE) has expanded the list of particles separable by use of gel electrophoresis to include: (1) linear DNAs as long as 3-6 Mbp, (2) DNA- protein complexes and circular DNAs that become arrested during invariant field agarose gel electrophoresis; and (3) micron-sized spheres that also become arrested during invariant field agarose gel electrophoresis (reviewed in refs. 1-3). During the replication, recombination and packaging of DNA by the various double-stranded DNA bacteriophages, circular DNAs, protein-DNA complexes, and end-to- end joined mature DNA multimers (concatemers) as long as 0.5-1.0 Mbp are formed (reviewed in refs. 4,5). Thus, in addition to being useful for genome mapping (1,2,6,7 and see also Chapter 18 ), PFGE also is useful for studying the DNA metabolism of both bacteriophages and, presumably, other viruses.

摘要

脉冲场琼脂糖凝胶电泳（PFGE）的引入，已将可通过凝胶电泳分离的颗粒种类扩展到包括：（1）长达3 - 6兆碱基对（Mbp）的线性DNA；（2）在恒定电场琼脂糖凝胶电泳过程中滞留的DNA - 蛋白质复合物和环状DNA；以及（3）在恒定电场琼脂糖凝胶电泳过程中也会滞留的微米级球体（参考文献1 - 3中有综述）。在各种双链DNA噬菌体对DNA进行复制、重组和包装的过程中，会形成环状DNA、蛋白质 - DNA复合物以及长达0.5 - 1.0兆碱基对的端对端连接的成熟DNA多联体（串联体）（参考文献4、5中有综述）。因此，PFGE除了可用于基因组图谱绘制（参考文献1、2、6、7，另见第18章）外，还可用于研究噬菌体以及可能的其他病毒的DNA代谢。