生物
质粒
微生物学
DNA测序
阴道加德纳菌
乳酸菌
转化(遗传学)
细菌
计算生物学
基因
遗传学
细菌性阴道病
作者
Elsa Fristot,Thomas Bessède,Miguel Camacho Rufino,Pauline Mayonove,Hao-Hsun Chang,Ana Zúñiga,Anne-Laure Michon,Sylvain Godreuil,J. Bonnet,Guillaume Cambray
出处
期刊:PLOS ONE
[Public Library of Science]
日期:2023-02-17
卷期号:18 (2): e0280935-e0280935
被引量:1
标识
DOI:10.1371/journal.pone.0280935
摘要
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 3.5·103 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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