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

Replication of circular and linear SV40-based plasmids in monkey cells

F Ascenzioni1, L Pucci, A M Guerrini

  • 1Istituto Pasteur-Fondazione Cenci Bolognetti, Dipartimento di Biologia Cellulare e dello Sviluppo, Università La Sapienza, Roma, Italy.

Research in Virology
|September 1, 1996
PubMed
Summary
This summary is machine-generated.

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

Hypnosis and superficial cervical anesthesia for total thyroidectomy in a high-risk patient - A case report.

International journal of surgery case reports·2020
Same author

Listeriolysin O antibodies detection in pregnant women: results from an Italian pilot study.

Annali di igiene : medicina preventiva e di comunita·2019
Same author

Municipal wastewater spiramycin removal by conventional treatments and heterogeneous photocatalysis.

The Science of the total environment·2017
Same author

Blood and urine inducible protein 10 as potential markers of disease activity.

The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease·2016
Same author

Prevention of vascular damage with Lisosan G wheat extract: the in vitro basis for a clinical investigation.

European review for medical and pharmacological sciences·2015
Same author

Cytogenetic and molecular mapping of the wheat-Aegilops longissima chromatin breakpoints in powdery mildew-resistant introgression lines.

TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·2013

Replication of SV40-based plasmids in COS-7 cells depends on DNA structure. Circular plasmids replicate, while linear forms do not, though circularization restores replication potential.

Area of Science:

  • Molecular Biology
  • Virology
  • Genetics

Background:

  • SV40 (Simian Vacuolating Virus 40) origin of replication (ori) is crucial for viral DNA replication.
  • Telomeric sequences play a role in DNA stability and replication.
  • Understanding plasmid replication mechanisms is vital for gene delivery and biotechnology.

Purpose of the Study:

  • To investigate the impact of DNA structure (circular vs. linear) and telomeric sequences on SV40-based plasmid replication.
  • To assess the functional integrity of the SV40 ori in different plasmid forms.
  • To elucidate the role of host cell factors in viral DNA replication.

Main Methods:

  • Construction of circular (pYACneoC) and linear (pYACneoL, pYACneoN) SV40-based plasmids with varying telomeric sequences.

Related Experiment Videos

  • Transfection of COS-7 cells with the constructed plasmids.
  • Extraction and analysis of low-molecular-weight DNA at various time points post-transfection.
  • Assessment of replication by DpnI digestion to quantify replicated molecules.

    • Main Results:

    • Circular plasmids (pYACneoC) showed replication in approximately 50% of molecules shortly after transfection.
    • Linear plasmids (pYACneoL and pYACneoN) did not exhibit replication.
    • Circularized forms recovered from linear plasmids demonstrated replication, with pYACneoL showing higher efficiency than pYACneoN.
    • Replication of circular plasmids was transient, occurring only in a narrow window post-transfection.

    Conclusions:

    • Plasmid DNA structure significantly influences SV40 ori-mediated replication.
    • Linear DNA topology inhibits replication, despite the presence of a functional SV40 ori.
    • Host cell functions and DNA conformation are critical determinants of viral replication efficiency.