Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Autonomous replication of plasmids bearing monkey DNA origin-enriched sequences.

L Frappier1, M Zannis-Hadjopoulos

  • 1McGill Cancer Centre, Department of Medicine, McGill University, Montreal, Quebec, Canada.

Proceedings of the National Academy of Sciences of the United States of America
|October 1, 1987
PubMed
Summary

Four monkey origin-enriched sequences (ORS) support plasmid replication in mammalian cells, demonstrating controlled, semiconservative replication. This finding advances understanding of DNA replication origins.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Replication initiation and DNA topology: The twisted life of the origin.

Journal of cellular biochemistry·2010
Same author

Dynamic changes in chromatin structure through post-translational modifications of histone H3 during replication origin activation.

Journal of cellular biochemistry·2009
Same author

Metazoan origins of DNA replication: regulation through dynamic chromatin structure.

Journal of cellular biochemistry·2009
Same author

HAUSP/USP7 as an Epstein-Barr virus target.

Biochemical Society transactions·2004
Same author

14-3-3s are DNA-replication proteins.

Biochemical Society transactions·2002
Same author

In vivo association of Ku with mammalian origins of DNA replication.

Molecular biology of the cell·2001

Area of Science:

  • Molecular Biology
  • Genetics
  • Mammalian Cell Culture

Background:

  • Early replicating DNA sequences, termed origin-enriched sequences (ORS), are crucial for understanding DNA replication initiation.
  • Investigating the function of monkey ORS in heterologous mammalian cell systems can reveal conserved replication mechanisms.

Purpose of the Study:

  • To identify and characterize monkey ORS capable of supporting transient episomal replication in various mammalian cell lines.
  • To determine the mode of replication supported by these ORS and their efficiency across different cell types.

Main Methods:

  • Isolation and cloning of twelve ORS from early replicating monkey (CV-1) DNA.
  • Transient transfection of CV-1, COS-7, and HeLa cells with ORS-containing plasmids.
  • Analysis of plasmid replication using Dpn I resistance and bromodeoxyuridine substitution assays.

Related Experiment Videos

  • Electron microscopy for visualization of replication intermediates and restriction mapping of replication bubbles.

    • Main Results:

    • Four monkey ORS (ORS3, -8, -9, and -12) were identified as capable of supporting plasmid replication.
    • Replication was demonstrated to be controlled and semiconservative, consistent with mammalian replicon activity.
    • Replication efficiency varied, with HeLa cells showing the highest activity; electron microscopy confirmed replication bubbles in ORS8 and ORS12.
    • Replication bubble mapping localized the initiation site within the ORS12 sequence.

    Conclusions:

    • Monkey ORS can function as functional replication origins in diverse mammalian cell environments.
    • The identified ORS support a canonical mode of DNA replication, offering valuable tools for studying replication.
    • HeLa cells provide a highly efficient system for studying monkey ORS-driven replication.