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Related Experiment Videos

Replication of prophage P1 is cell-cycle specific.

J D Keasling1, B O Palsson, S Cooper

  • 1Department of Chemical Engineering, University of Michigan, Ann Arbor 48109-2136.

Journal of Bacteriology
|July 1, 1992
PubMed
Summary

The P1 prophage replicates during a specific part of the Escherichia coli cell cycle, distinct from the minichromosome. This replication timing suggests similar mechanisms to other plasmids and the bacterial chromosome.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Bacterial Genetics

Background:

  • Bacteriophage P1 is a temperate phage that integrates into the Escherichia coli genome as a prophage.
  • Understanding the replication timing of P1 prophage is crucial for comprehending its lifecycle and host-phage interactions within the bacterial division cycle.

Purpose of the Study:

  • To investigate the temporal regulation of P1 prophage replication during the Escherichia coli division cycle.
  • To compare the replication timing of P1 prophage with other genetic elements, such as the minichromosome and a high-copy plasmid, within the same bacterial cell.

Main Methods:

  • Utilized the membrane-elution technique to isolate and label Escherichia coli cells at precise stages of their division cycle.
  • Employed scintillation counting for quantitative analysis of radioactive P1 prophage DNA, enabling precise measurement of replication events.

Main Results:

  • P1 prophage replication occurs during a restricted interval of the bacterial division cycle.
  • P1 prophage replication is temporally separated from minichromosome replication, occurring approximately half a generation later.
  • A high-copy mini-R6K plasmid replicates continuously throughout the division cycle, contrasting with the regulated replication of P1 prophage.

Conclusions:

  • P1 prophage replication is tightly regulated and linked to specific cellular events, similar to chromosomal and F plasmid replication.
  • Replication initiation for P1 prophage appears to be dependent on reaching a critical cell size or mass per origin.
  • The distinct replication timing suggests unique regulatory mechanisms governing P1 prophage DNA synthesis within the host cell.

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