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

A new site-specific integration system for mycobacteria.

Jeffrey Murry1, Christopher M Sassetti, Jonathan Moreira

  • 1Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.

Tuberculosis (Edinburgh, Scotland)
|November 1, 2005
PubMed
Summary

The Streptomyces phiC31 integration system enables site-specific DNA integration into mycobacterial chromosomes, creating stable genetic tools. This method offers enhanced stability for studying mycobacterial genetics and pathogenesis.

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

Ubiquitylation by the GID/CTLH complex regulates the metabolic and innate immune response of macrophages to infection by <i>Mycobacterium tuberculosis</i>.

bioRxiv : the preprint server for biology·2026
Same author

Cross-species blood transcriptional correlates of BCG-mediated protection against tuberculosis include innate and adaptive immune processes.

JCI insight·2025
Same author

Reference-based chemical-genetic interaction profiling to elucidate small molecule mechanism of action in Mycobacterium tuberculosis.

Nature communications·2025
Same author

Cross-species blood transcriptional correlates of BCG-mediated protection against tuberculosis include innate and adaptive immune processes.

bioRxiv : the preprint server for biology·2025
Same author

The Mycobacterium tuberculosis Transposon Sequencing Database (MtbTnDB): A Large-Scale Guide to Genetic Conditional Essentiality.

Molecular microbiology·2025
Same author

Antibody-Fab and -Fc features promote Mycobacterium tuberculosis restriction.

Immunity·2025

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Site-specific integration into the mycobacterial chromosome is crucial for stable transformants.
  • Gene expression challenges can arise at specific integration sites.
  • Understanding mycobacterial pathogenesis requires stable genetic manipulation tools.

Purpose of the Study:

  • To utilize the Streptomyces phiC31 integration system for site-specific DNA integration in mycobacteria.
  • To assess the stability and efficiency of this integration system across different mycobacterial species.
  • To evaluate the utility of phiC31-based plasmids as tools for mycobacterial genetics research.

Main Methods:

  • Employing the Streptomyces phiC31 integration system for site-specific recombination.

Related Experiment Videos

  • Integrating vector DNA into Mycobacterium smegmatis, Mycobacterium bovis BCG, and Mycobacterium tuberculosis.
  • Analyzing insertion sites and plasmid stability through in vitro passage.

    • Main Results:

    • A single dominant insertion site was identified in Mycobacterium smegmatis.
    • Three distinct insertion sites were observed in Mycobacterium bovis BCG.
    • Integrated vectors in Mycobacterium smegmatis demonstrated greater stability than episomal plasmids during unselected passage.

    Conclusions:

    • The phiC31 integration system is effective for site-specific DNA integration in various mycobacteria.
    • phiC31-based integration offers enhanced stability for genetic elements in mycobacteria compared to plasmids.
    • phiC31 integration system plasmids represent valuable tools for advancing mycobacterial genetics and pathogenesis studies.