An optimized electrotransformation protocol for Lactobacillus jensenii
Open accessAbstract
Engineered bacteria are promising candidates for in situ detection and treatment of diseases. The female uro-genital tract presents several pathologies, such as sexually transmitted diseases or genital cancer, that could benefit from such technology. While bacteria from the gut microbiome are increasingly engineered, the use of chassis isolated from the female uro-genital resident flora has been limited. A major hurdle to implement the experimental throughput required for efficient engineering in these non-model bacteria is their low transformability. Here we report an optimized electrotransformation protocol for Lactobacillus jensenii , one the most widespread species across vaginal microflora. Starting from classical conditions, we optimized buffers, electric field parameters, cuvette type and DNA quantity to achieve an 80-fold improvement in transformation efficiency, with up to 3.5·10 3 CFUs/μg of DNA in L . jensenii ATCC 25258 . We also identify several plasmids that are maintained and support reporter gene expression in L . jensenii . Finally, we demonstrate that our protocol provides increased transformability in three independent clinical isolates of L . jensenii . This work will facilitate the genetic engineering of L . jensenii and enable its use for addressing challenges in gynecological healthcare.
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L. jensenii is present in approximately 20% of women, suggesting a larger cohort would have been necessary for its isolation.
“L. jensenii, while typical of a healthy vaginal microbiota, is present in approximately 20% of the women, suggesting that a larger cohort would have been necessary for its isolation [48, 49].”
From Women's health is a team effort: probiogenomics supports the development of a multi-species vaginal probiotic (2026) · cites this paper as [48]
The Introduction (page not provided) describes L. jensenii as a vaginal resident and says it represents approximately 20% of vaginal flora in most groups studied. That is not the same as being present in 20% of women. The paper does not say that a larger cohort would have been necessary to isolate it. (LLM, conf=0.86)
Experimental — accuracy not yet measuredLm. fermentum is not typically a dominant species in the gastrointestinal tract or vagina, but its established probiotic characteristics make it a promising candidate.
“WhileLm. fermentumis not typically a dominant species in the gastrointestinal tract or in the vagina, its established probiotic characteristics make it a promising candidate [48, 50–52].”
From Women's health is a team effort: probiogenomics supports the development of a multi-species vaginal probiotic (2026) · cites this paper as [48]
The Introduction discusses probiotic potential for lactobacilli generally and identifies L. jensenii as a candidate for vaginal microbiome engineering. A search for “fermentum” found only a plasmid-origin entry in the Results table; our copy does not address Lm. fermentum’s abundance in the gastrointestinal tract or vagina, or its probiotic characteristics. (LLM, conf=0.74)
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References 26 · Cited by 9
Research works printed but not linked: 1
- [4]Ruzal SM. Lactobacillus Genomics and Metabolic Engineering. Ruzal S, editor. Poole, England: Caister Academic Press; 2019. 222 p.
- Authors
- Fristot, E., Bessede, T., Camacho Rufino, M., Mayonove, P., Chang, H.-J., Zuniga, A., Michon, A.-L., Godreuil, S., Bonnet, J., & Cambray, G.
- Affiliations
- Centre National de la Recherche Scientifique (FR)
- Université de Montpellier (FR)
- Centre Hospitalier Universitaire de Montpellier (FR)
- Corresponding author
- Bonnet, Jérôme
- Cambray, Guillaume
- Venue
- PLOS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2023-02-17
- Volume
- 18
- Issue
- 2
- Pages
- e0280935
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC9937494
- OpenAlex
- W4321166976
- Cited by
- 9
- References
- 26
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 1 · 2023: 2
- FWCI
- 1.49
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- PLoS ONE (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- HAL (submittedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- ATIP-Avenir
- Fondation Bettencourt Schueller
- Ministère de l'Enseignement Supérieur et de la Recherche
- Ligue Contre le Cancer
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Immunology and Microbiology › Microbiology › Reproductive tract infections research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Keywords
- Lactobacillus jensenii, electrotransformation, transformation efficiency optimization, vaginal microbiota, urogenital tract, bacterial chassis, plasmid maintenance, reporter gene expression, clinical isolates, gynecological diseases
- MeSH terms
- Bacteria, Female, Humans, Lactobacillus, Plasmids, Vagina
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