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

DNA recombination during PCR.

A Meyerhans1, J P Vartanian, S Wain-Hobson

  • 1Laboratoire de Biologie et Immunologie Moléculaires des Rétrovirus, Institut Pasteur, Paris, France.

Nucleic Acids Research
|April 11, 1990
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

[Mathematical Modeling of the Intracellular Regulation of Immune Processes].

Molekuliarnaia biologiia·2019
Same author

Cancer immunotherapy of patients with HIV infection.

Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico·2018
Same author

Interplay between reaction and diffusion processes in governing the dynamics of virus infections.

Journal of theoretical biology·2018
Same author

Towards standardized automated immunomonitoring: an automated ELISpot assay for safe and parallelized functionality analysis of immune cells.

Cytotechnology·2016
Same author

APOBEC3A intratumoral DNA electroporation in mice.

Gene therapy·2016
Same author

Detection of Broadly Neutralizing Activity within the First Months of HIV-1 Infection.

Journal of virology·2016

Polymerase chain reaction (PCR) can create new DNA by combining sequences. Increasing Taq DNA polymerase time reduces unwanted recombination, important for studying genetic material like RNA viruses.

Area of Science:

  • Molecular Biology
  • Genetics
  • Virology

Background:

  • Polymerase chain reaction (PCR) is a key technique for DNA amplification.
  • Recombination during PCR can complicate analysis of genetic material.
  • The HIV1 tat gene is a target for studying viral gene expression.

Purpose of the Study:

  • To investigate PCR-mediated recombination between distinct HIV1 tat gene sequences.
  • To determine factors influencing the frequency of PCR recombination.
  • To explore the potential applications of PCR recombination.

Main Methods:

  • Co-amplification of two distinct HIV1 tat gene sequences using PCR.
  • Analysis of amplified DNA molecules for recombinant formation.
  • Mapping of crossover sites within recombinant DNA.

Related Experiment Videos

  • Varying Taq DNA polymerase elongation time to assess its effect on recombination.

    • Main Results:

    • PCR co-amplification resulted in the formation of recombinant DNA molecules.
    • Recombinant frequencies reached up to 5.4% of amplified molecules.
    • Increasing Taq DNA polymerase elongation time by 6-fold decreased recombinant frequency by 2.7-fold.
    • Crossover sites were localized to three specific regions, suggesting polymerase pause/termination sites.

    Conclusions:

    • PCR-mediated recombination occurs between distinct gene sequences.
    • Taq DNA polymerase elongation time is a critical factor in controlling PCR recombination.
    • This recombination phenomenon can be a challenge for analyzing heterogeneous genetic material.
    • PCR recombination can be harnessed to generate chimeric DNA molecules.