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

Stable low-copy-number Staphylococcus aureus shuttle vectors.

Steve Grkovic1, Melissa H Brown1, Kate M Hardie1

  • 1School of Biological Sciences, Macleay Building A12, University of Sydney, New South Wales 2006, Australia.

Microbiology (Reading, England)
|March 14, 2003
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

A large retrospective study on 1PN embryo transfer supports the need for updated harmonized best practice guidelines.

Human reproduction (Oxford, England)·2026
Same author

Efficacy of physical and chemical disinfection against clinically relevant free-living amoebae isolated from drinking water and plumbing biofilms.

Frontiers in microbiology·2025
Same author

Is CepA from <i>Klebsiella pneumoniae</i> a biocide pump? Evidence suggests a metal efflux function.

mSphere·2025
Same author

Handwashing basins and healthcare associated infections: Bacterial diversity in biofilms on faucets and drains.

The Science of the total environment·2024
Same author

Increased flushing frequency of a model plumbing system initially promoted the formation of viable but non culturable cells but ultimately reduced the concentration of culturable and total <i>Legionella</i> DNA.

Heliyon·2024
Same author

pSK41/pGO1-family conjugative plasmids of Staphylococcus aureus encode a cryptic repressor of replication.

Plasmid·2023

New Staphylococcus aureus-Escherichia coli shuttle vectors, based on the pSK1 plasmid, enable stable, low-copy-number gene expression and promoter analysis in S. aureus, overcoming common stability issues.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • Staphylococcus aureus gene expression studies are often hindered by vector instability.
  • Existing Gram-positive vectors, particularly rolling-circle types, suffer from segregational and structural instability.
  • Efficient tools for analyzing gene function and regulation in S. aureus are needed.

Purpose of the Study:

  • To construct and validate novel Staphylococcus aureus-Escherichia coli shuttle vectors.
  • To improve gene cloning and expression efficiency in S. aureus.
  • To develop tools for promoter strength determination and gene analysis at low copy number.

Main Methods:

  • Construction of shuttle vectors incorporating S. aureus theta-mode replication functions from plasmid pSK1.

Related Experiment Videos

  • Cloning and expression of previously difficult-to-express genes in S. aureus.
  • Generation of vectors for transcriptional and translational fusions using the S. aureus blaZ reporter gene.
  • Assessment of promoter strengths in S. aureus.

    • Main Results:

    • Successfully constructed and demonstrated the utility of S. aureus-E. coli shuttle vectors.
    • Achieved efficient cloning and expression of challenging genes in S. aureus.
    • Developed reporter gene fusion vectors for promoter analysis.
    • Demonstrated enhanced stability and low-copy-number replication, avoiding common issues with rolling-circle vectors.

    Conclusions:

    • The new pSK1-based shuttle vectors offer improved stability for gene expression and analysis in Staphylococcus aureus.
    • These vectors facilitate low-copy-number studies, overcoming limitations of previous systems.
    • The developed tools are valuable for investigating gene function and regulation in S. aureus.