Conjugative DNA Transfer From E. coli to Transformation-Resistant Lactobacilli
| Autor: | Miguel A. Alvarez, Dolores L. Guzmán-Herrador, Sara Samperio, Matxalen Llosa, Rigoberto May-Cuz, Maria Cruz Martin |
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| Přispěvatelé: | Universidad de Cantabria, Ministerio de Economía y Competitividad (España), Agencia Estatal de Investigación (España), European Commission |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Microbiology (medical)
plasmid R388 lcsh:QR1-502 medicine.disease_cause Microbiology lcsh:Microbiology 03 medical and health sciences chemistry.chemical_compound Plasmid Staphylococcus epidermidis Lactobacillus medicine Lactic acid bacteria Escherichia coli plasmid RP4 030304 developmental biology Original Research 0303 health sciences biology Plasmid RP4 030306 microbiology Bacterial conjugation biology.organism_classification Plasmid R388 lactic acid bacteria Transformation (genetics) bacterial conjugation chemistry Bacteria DNA |
| Zdroj: | Frontiers in Microbiology Front Microbiol . 2021 Feb 11;12:606629 Digital.CSIC: Repositorio Institucional del CSIC Consejo Superior de Investigaciones Científicas (CSIC) UCrea Repositorio Abierto de la Universidad de Cantabria Universidad de Cantabria (UC) Frontiers in Microbiology, Vol 12 (2021) Digital.CSIC. Repositorio Institucional del CSIC instname |
| ISSN: | 1664-302X 2017-8719 |
| Popis: | © 2021 Samperio, Guzmán-Herrador, May-Cuz, Martín, Álvarez and Llosa. 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. Work in ML lab was supported by the grant BIO2017-87190-R from the Spanish Ministry of Science and Innovation. Work in MÁ lab was funded by the Spanish State Research Agency (AEI) and the European Regional Development Fund (FEDER) (AGL2016-78708-R, AEI/FEDER, EU). DG-H was a recipient of a predoctoral appointment from the University of Cantabria. RM-C received an Erasmus+ traineeship grant. |
| Databáze: | OpenAIRE |
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