ArticleNucleic acids research2012
High efficiency recombineering in lactic acid bacteria.
Article in Nucleic acids research, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 127 papers.
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Who cites it
127 citing papers in PubMed, 227 citations in OpenAlex.
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- Genetic Toolbox Expansion Enables Constitutively Fluorescent Lacticaseibacillus rhamnosus for Functional Microbiome Research.Microbial biotechnology · 2026Article
- Genetically Modified Lactic Acid Bacteria in the EU Food Chain: Applications, Benefits, and Risk Assessment.International journal of molecular sciences · 2026Review
- Genome editing of phylogenetically distinct bacteria using cross-species retron-mediated recombineering.Nature biotechnology · 2026Article
- Enhanced editing ofApplied and environmental microbiology · 2026Article
- Advances in gene editing tools for four typical Gram-positive bacteria.Frontiers in microbiology · 2026Review
- Being a better version of yourself: genetically engineered probiotic bacteria as host defense enhancers in the control of intestinal pathogens.Gut microbes · 2025Review
- Engineered Lactiplantibacillus plantarum and Levilactobacillus brevis utilizing ribonucleoprotein-mediated editing for inactivation of hemolysin gene.World journal of microbiology & biotechnology · 2025Article
- Unlocking Casein Bioactivity: Lactic Acid Bacteria and Molecular Strategies for Peptide Release.International journal of molecular sciences · 2025Review
- Genome-Editing Tools for Lactic Acid Bacteria: Past Achievements, Current Platforms, and Future Directions.International journal of molecular sciences · 2025Review
- A diverse single-stranded DNA-annealing protein library enables efficient genome editing across bacterial phyla.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Development of aApplied and environmental microbiology · 2024Article
- Functional and practical insights into three lactococcal antiphage systems.Applied and environmental microbiology · 2024Article
- TheApplied and environmental microbiology · 2024Article
- Half a century after their discovery: Structural insights into exonuclease and annealase proteins catalyzing recombineering.Engineering microbiology · 2024Review
- New Genomic Techniques applied to food cultures: a powerful contribution to innovative, safe, and sustainable food products.FEMS microbiology letters · 2024Article
- Discovering genetic determinants for cell-to-cell adhesion in two prevalent conjugative lactococcal plasmids.Current research in microbial sciences · 2024Article
- Molecular mechanisms underlying the structural diversity of rhamnose-rich cell wall polysaccharides in lactococci.The Journal of biological chemistry · 2024Article
- Transcriptional control of two distinct lactococcal plasmid-encoded conjugation systems.Current research in microbial sciences · 2024Article
67 more citing papers are in PubMed but not listed here.
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
The ability to efficiently generate targeted point mutations in the chromosome without the need for antibiotics, or other means of selection, is a powerful strategy for genome engineering. Although oligonucleotide-mediated recombineering (ssDNA recombineering) has been utilized in Escherichia coli for over a decade, the successful adaptation of ssDNA recombineering to gram-positive bacteria has not been reported. Here we describe the development and application of ssDNA recombineering in lactic acid bacteria. Mutations were incorporated in the chromosome of Lactobacillus reuteri and Lactococcus lactis without selection at frequencies ranging between 0.4% and 19%. Whole genome sequence analysis showed that ssDNA recombineering is specific and not hypermutagenic. To highlight the utility of ssDNA recombineering we reduced the intrinsic vancomymycin resistance of L. reuteri >100-fold. By creating a single amino acid change in the D-Ala-D-Ala ligase enzyme we reduced the minimum inhibitory concentration for vancomycin from >256 to 1.5 µg/ml, well below the clinically relevant minimum inhibitory concentration. Recombineering thus allows high efficiency mutagenesis in lactobacilli and lactococci, and may be used to further enhance beneficial properties and safety of strains used in medicine and industry. We expect that this work will serve as a blueprint for the adaptation of ssDNA recombineering to other gram-positive bacteria.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.