Conjugative DNA Transfer From E. coli to Transformation-Resistant Lactobacilli
Open accessAbstract
Lactic acid bacteria (LAB) belonging to the genus classically known as Lactobacillus , recently split into 25 different genera, include many relevant species for the food industry. The well-known properties of lactobacilli as probiotics make them an attractive model also for vaccines and therapeutic proteins delivery in humans. However, scarce tools are available to accomplish genetic modification of these organisms, and most are only suitable for laboratory strains. Here, we test bacterial conjugation as a new tool to introduce genetic modifications into many biotechnologically relevant laboratory and wild type lactobacilli. Using mobilizable shuttle plasmids from a donor Escherichia coli carrying either RP4 or R388 conjugative systems, we were able to get transconjugants to all tested Lactocaseibacillus casei strains, including many natural isolates, and to several other genera, including Lentilactobacillus parabuchneri , for which no transformation protocol has been reported. Transconjugants were confirmed by the presence of the oriT and 16S rRNA gene sequencing. Serendipitously, we also found transconjugants into researcher-contaminant Staphylococcus epidermidis . Conjugative DNA transfer from E. coli to S. aureus was previously described, but at very low frequencies. We have purified this recipient strain and used it in standard conjugation assays, confirming that both R388 and RP4 conjugative systems mediate mobilization of plasmids into S. epidermidis . This protocol could be assayed to introduce DNA into other Gram-positive microorganisms which are resistant to transformation.
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Risk categories for probiotics are associated with four interrelated factors: acquired antimicrobial resistance genes, virulence factors, translocation, and mobile or unstable genomic elements that allow horizontal gene transfer.
“Risk categories for probiotics are associated with four interrelated factors: acquired antimicrobial resistance genes, virulence factors, translocation, and mobile or unstable genomic elements that allow horizontal gene transfer [43,44].”
From Current Concepts in Probiotic Safety and Efficacy (2026) · cites this paper as [44]
The Introduction (page unavailable) says conjugation is a means of spreading resistance and virulence factors among bacteria, and the Discussion reports plasmid transfer into lactobacilli. The paper does not set out four probiotic risk categories or address translocation and unstable genomic elements as parts of that framework. (LLM, conf=0.84)
Experimental — accuracy not yet measuredIn vitro and model-system experimental studies have shown plasmid conjugation between enterobacteria and Gram-positive recipients, including LAB.
“In vitro and in model systems, experimental studies have shown plasmid conjugation between enterobacteria and Gram-positive recipients, including LAB [44,123].”
From Current Concepts in Probiotic Safety and Efficacy (2026) · cites this paper as [44]
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- Authors
- Samperio, S., Guzmán-Herrador, D. L., May-Cuz, R., Martín, M. C., Álvarez, M. A., & Llosa, M.
- Affiliations
- Universidad de Cantabria (ES)
- Consejo Superior de Investigaciones Científicas (ES)
- Corresponding author
- Álvarez, Miguel A.
- Llosa, Matxalen
- Venue
- Frontiers in Microbiology
- Publisher
- Frontiers Media SA
- Published
- 2021-02-11
- Volume
- 12
- Pages
- 606629-606629
- ISSN
- 1664-302X (ISSN-L)
- PMC
- PMC7905204
- OpenAlex
- W3129747008
- Cited by
- 25
- References
- 80
- Citations by year
- 2026: 5 · 2025: 10 · 2024: 5 · 2023: 3 · 2022: 2
- FWCI
- 1.41
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Frontiers in Microbiology (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DIGITAL.CSIC (Spanish National Research Council (CSIC)) (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- UCrea (University of Cantabria) (submittedVersion, cc-by)
- Funders
- Ministerio de Ciencia e Innovación
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Agricultural and Biological Sciences › Food Science › Probiotics and Fermented Foods
- Physical Sciences › Environmental Science › Ecology › Bacteriophages and microbial interactions
- Keywords
- bacterial conjugation, conjugative transfer, Lactobacillus, Lentilactobacillus parabuchneri, Staphylococcus epidermidis, Staphylococcus aureus, plasmid mobilization, probiotics
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