Study of plasmid replication in Escherichia coli with a combination of 2D gel electrophoresis and electron microscopy

Andrei Kuzminov, Eric Schabtach, Franklin W. Stahl

Research output: Contribution to journalEditorialpeer-review

Abstract

We studied θ-mode DNA replication in p15A-based Escherichia coli plasmids by analyzing their replication intermediates using a combination of neutral agarose 2D gel electrophoresis and electron microscopy. Our analysis: (1) confirms the original assignment of various features of the 2D gel pattern; (2) shows that while one replication fork progresses around the plasmid DNA, the other is immobile, as if the replication were unidirectional; and (3) reveals that termination often occurs at a location away from the replication origin, suggesting that the replication of our plasmids is, in fact, bidirectional, the two forks being active at different times.

Original languageEnglish (US)
Pages (from-to)1-7
Number of pages7
JournalJournal of Molecular Biology
Volume268
Issue number1
DOIs
StatePublished - Apr 25 1997
Externally publishedYes

Keywords

  • 2D agarose gel-electrophoresis
  • Electron microscopy
  • P15A replicon
  • Replication intermediates
  • Unidirectional replication

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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