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Bidirection replication from a unique origin in a mini-F plasmid.

R Eichenlaub, D Figurski, D R Helinski

    Proceedings of the National Academy of Sciences of the United States of America
    |March 1, 1977
    PubMed
    Summary
    This summary is machine-generated.

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    Replication of the mini F-kanamycin resistance plasmid (pML31) in Escherichia coli primarily occurs bidirectionally from a specific origin. Termination sites are located opposite this origin, near the kanamycin resistance gene.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • The mini F-kanamycin resistance plasmid, pML31, is a derivative of the F'lac plasmid.
    • Understanding plasmid replication mechanisms is crucial for genetic engineering and molecular biology.

    Purpose of the Study:

    • To investigate the replication mode and origin of the pML31 plasmid in Escherichia coli.
    • To determine the replication termination sites of pML31.

    Main Methods:

    • Isolation of covalently-closed circular DNA molecules of pML31 from Escherichia coli.
    • Electron microscopy analysis of DNA molecules after digestion with EcoRI and BamHI restriction endonucleases.

    Main Results:

    • The majority of pML31 molecules exhibited bidirectional replication, initiating from a unique origin on the F-fragment.

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  • This origin was mapped to 42.6 kilobases on the F-fragment, approximately 2.3 kilobases from an EcoRI site.
  • A small fraction of molecules showed unidirectional replication from the origin.
  • Replication termination was observed in a unique region opposite the origin, within the fragment containing the kanamycin resistance locus.

    • Conclusions:

    • Plasmid pML31 replicates via a predominantly bidirectional mechanism in Escherichia coli.
    • The identified origin and termination sites provide insights into the control of plasmid DNA replication.