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Journal of Biological Sciences

Year: 2005 | Volume: 5 | Issue: 2 | Page No.: 222-229
DOI: 10.3923/jbs.2005.222.229

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Authors


Stephen Cooper


Jay D. Keasling


Keywords


  • E. coli
  • plasmid replication
  • membrane-elution
  • Double- label
  • p1
Research Article

Experimental and Theoretical Considerations of P1-plasmid Replication and Segregation During the E. coli Cell Cycle

Stephen Cooper and Jay D. Keasling
Two contrasting experimental descriptions of P1-plasmid replication during the cell cycle of Escherichia coli have been described. One set of results led to the proposal that replication of P1-plasmid occurs at a specific time during the cell cycle over a wide range of growth rates and follows rules similar to that governing bacterial chromosome replication. Experiments supporting this proposal utilized membrane-elution experiments, radioactive double-labeling of DNA and scintillation counting of purified plasmids. An alternative experimental description of P1-plasmid replication during the cell cycle, also based on membrane-elution methodology but measuring radioactivity incorporated into plasmid DNA by autoradiography and scanning of films, proposed that P1-plasmid replicates at all stages of the cell cycle in rapidly growing cells, but with a slight periodicity or increase in P1 replication probability within the cell cycle of slower growing cells. These discordant experimental results are analyzed. It is concluded that the direct double-label counting approach is to be preferred, as the results are consistent with a large number of experiments, are supported by theoretical considerations and yield a unified view of plasmid replication over a wide range of growth rates. Theoretical ramifications of each view of P1-plasmid replication-cycle-dependent and cycle-independent are compared. An analysis of P1-plasmid segregation is also presented.
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How to cite this article

Stephen Cooper and Jay D. Keasling, 2005. Experimental and Theoretical Considerations of P1-plasmid Replication and Segregation During the E. coli Cell Cycle. Journal of Biological Sciences, 5: 222-229.

DOI: 10.3923/jbs.2005.222.229

URL: https://scialert.net/abstract/?doi=jbs.2005.222.229

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