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 theta-type thermosensitive replicon from Lactococcus lactis as an integration vector for Enterococcus faecalis

J Frère1, A Benachour, J C Giard

  • 1Laboratoire de Microbiologie de l'Environnement, IRBA, Université de Caen, France. frere@ibba.unicaen.fr

FEMS Microbiology Letters
|April 30, 1998
PubMed
Summary

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

[Impact of COVID-19 outbreak on domestic violence and child abuse].

Revue medicale de Liege·2021
Same author

The sponges Hymeniacidon perlevis and Halichondria panicea are reservoirs of antibiotic-producing bacteria against multi-drug resistant Staphylococcus aureus.

Journal of applied microbiology·2021
Same author

Diversity and dynamics of bacterial and fungal communities in cider for distillation.

International journal of food microbiology·2020
Same author

[Cat-scratch disease].

Revue medicale de Liege·2020
Same author

Characterization of Putative Virulence Factors of <i>Pseudomonas aeruginosa</i> Strain RBS Isolated from a Saltern, Tunisia: Effect of Metal Ion Cofactors on the Structure and the Activity of LasB.

BioMed research international·2020
Same author

[Kingella kingae bone and joint infections].

Revue medicale de Liege·2019

Researchers developed a new thermosensitive replicon, pUCB3522Ts, for efficient genetic integration in Enterococcus faecalis. This tool demonstrates high practicality for chromosomal integration via homologous recombination.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Thermosensitive replicons are valuable tools for conditional gene expression and genetic manipulation in bacteria.
  • The lactococcal theta-type replicon pUCL22 is a basis for developing such tools.
  • Efficient tools for targeted chromosomal integration are crucial for bacterial genetics.

Purpose of the Study:

  • To develop an improved thermosensitive replicon for genetic manipulation in Enterococcus faecalis.
  • To create a tool for efficient and targeted chromosomal integration in E. faecalis.
  • To validate the utility of the developed replicon as a genetic integrative tool.

Main Methods:

  • Isolation and characterization of a thermosensitive mutant of the lactococcal pUCL22 replicon.

Related Experiment Videos

  • Fusion of the repA and repB genes to create an improved thermosensitive replicon, pUCB3522Ts.
  • Cloning of an internal fragment of the Enterococcus faecalis JH2-2 npr gene into pUCB3522Ts.
  • Monitoring chromosomal integration via homologous recombination by temperature shift from 30°C to 42°C.

    • Main Results:

    • An improved thermosensitive replicon, pUCB3522Ts, was constructed by fusing repA and repB genes.
    • The pUCB3522Ts plasmid exhibited high instability at 42°C in Enterococcus faecalis.
    • Targeted integration into the E. faecalis JH2-2 npr gene was achieved with 85% efficiency in analyzed clones.

    Conclusions:

    • The developed pUCB3522Ts thermosensitive replicon is a practical and efficient tool for targeted chromosomal integration in Enterococcus faecalis.
    • This tool facilitates genetic manipulation by enabling selection of integrated clones at a non-permissive temperature.
    • The study demonstrates the successful application of a novel thermosensitive integrative system in Gram-positive bacteria.