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May 1, 1972Journal of Bacteriology1,193 citationsOpen Access

Nature of Col E 1 Plasmid Replication in Escherichia coli in the Presence of Chloramphenicol

DCDon B. Clewell

Key Points

  • To investigate the replication of Col E1 plasmid in Escherichia coli in the presence of chloramphenicol.
  • Examined replication rates of Col E1 plasmid after chloramphenicol addition for 10 to 15 hours.
  • Utilized glucose-Casamino Acids medium to analyze the rate of plasmid DNA synthesis.
  • Measured the increase in Col E1 DNA copies per cell before and after chloramphenicol treatment.
  • Replication of Col E1 plasmid continues despite halted chromosomal DNA synthesis.
  • Increased replication rate reaches approximately eight times the average needed for DNA doubling per cell.
  • Col E1 DNA copies increase to about 3,000 per cell, a 125-fold rise over the normal 24 copies.

Abstract

The colicinogenic factor E(1) (Col E(1)) in Escherichia coli continues to replicate by a semiconservative mechanism in the presence of chloramphenicol (CAP) for 10 to 15 hr, long after chromosomal deoxyribonucleic acid (DNA) synthesis has terminated. Following CAP addition, the rate of synthesis of plasmid DNA gradually increases to an extent dependent on the medium employed. Within 2 to 4 hr after the addition of CAP, replication in a glucose-Casamino Acids medium approaches a maximum rate representing approximately eight times an average rate which would be required for a net doubling of DNA per cell in one generation. The number of copies of Col E(1) DNA molecules that accumulate under these conditions approaches about 3,000 copies per cell, representing a 125-fold increase over the normal level of 24 copies per cell. The system is particularly convenient for studying the mechanism of DNA replication.

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Cite This Study

Don B. Clewell (1972) studied this question.

synapsesocial.com/papers/6a0934651d1abd907d161487https://doi.org/10.1128/jb.110.2.667-676.1972
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