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Related Experiment Videos

Are single-stranded circles intermediates in plasmid DNA replication?

H te Riele, B Michel, S D Ehrlich

    The EMBO Journal
    |March 1, 1986
    PubMed
    Summary
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    Plasmid pHV33, containing pC194 and pBR322 DNA, exists in double- and single-stranded forms in E. coli. This suggests single-stranded DNA is a key intermediate in plasmid replication.

    Area of Science:

    • Molecular Biology
    • Genetics
    • Microbiology

    Background:

    • Plasmid pC194 exhibits double- and single-stranded DNA forms in Bacillus subtilis and Staphylococcus aureus.
    • Understanding plasmid replication mechanisms is crucial in molecular biology.

    Purpose of the Study:

    • To investigate the DNA forms of the hybrid plasmid pHV33 (pBR322/pC194) in Escherichia coli.
    • To determine the role of single-stranded DNA in plasmid replication.

    Main Methods:

    • Construction of the hybrid plasmid pHV33.
    • Replication studies in Escherichia coli.
    • Analysis of double- and single-stranded DNA forms.

    Main Results:

    • The hybrid plasmid pHV33 exists as both double- and single-stranded DNA in E. coli when pC194 replication functions are active.

    Related Experiment Videos

  • Single-stranded pHV33 DNA conversion to double-stranded DNA is initiated at the rriB site on pBR322.
  • The efficiency of this conversion impacts the plasmid's double-stranded copy number.

    • Conclusions:

    • Single-stranded DNA is a critical intermediate in the replication of plasmid pHV33.
    • The interplay between pBR322 and pC194 replication machinery influences plasmid copy number.