ArticleScientific reports2024
Optimizing phage-based mutant recovery and minimizing heat effect in the construction of transposon libraries in Staphylococcus aureus.
Article in Scientific reports, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- Profiling of genetic determinants required for fitness of community-associated methicillin-resistantMicrobiology spectrum · 2026Article
- Staphylococcus aureus's golden-yellow pigment staphyloxanthin: production enhancement, analytical characterization, and biological attributes.Microbial cell factories · 2026Review
- Article
- The Secondary Resistome of Methicillin-ResistantAntibiotics (Basel, Switzerland) · 2025Article
- Competitive fitness of Staphylococcus aureus against nasal commensals depends on biotin biosynthesis and acquisition.The ISME journal · 2025Article
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Authors and funding
3 authors.
Funding
Abstract
Staphylococcus aureus (S. aureus), particularly Methicillin-resistant S. aureus (MRSA), poses a significant global public health threat, necessitating advanced methodologies to enhance our understanding of this organism at the omics levels. This study introduces a refined protocol for constructing and curing high-density transposon mutant (tn-mutant) libraries in S. aureus, addressing the challenges associated with low transductant yields, and the complex genetic manipulation mechanism in Gram-positive bacteria. Our methodology employs a Himar1 transposon based on a two-plasmid system, leveraging Himar1's high insertional efficiency in AT-rich organisms. Enhanced transduction efficiency was achieved through chloramphenicol pre-treatment and the use of modified enriched media. Complementing this, an optimized plasmid curing procedure ensured a representative and stable tn-mutant library. The protocol was successfully applied to multiple S. aureus strains, demonstrating an increase in mutant recovery and reduced post-curing impact. The method offers a robust approach for Transposon Insertion Sequencing (TIS) applications in S. aureus, enabling deeper insights into survival, resistance, and pathogenicity mechanisms. This protocol holds a significant potential for accelerating the construction of tn-mutant libraries in various S. aureus strains.
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Registered trials
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