Development of a high-throughput platform to measure plasmid transfer frequency
dc.contributor.author | Arbé Cartón, Kepa | |
dc.contributor.author | Rey Sogo, Ana | |
dc.contributor.author | Santos Fernández, Nagore | |
dc.contributor.author | Altube, Oihane | |
dc.contributor.author | Garbisu Crespo, Carlos | |
dc.contributor.author | Arana Urbieta, Lide | |
dc.contributor.author | Alcorta Calvo, Miren Itziar | |
dc.date.accessioned | 2024-05-03T17:23:21Z | |
dc.date.available | 2024-05-03T17:23:21Z | |
dc.date.issued | 2023-10 | |
dc.identifier.citation | Frontiers in Cellular and Infection Microbiology 13 : (2023) // Article ID 1269732 | es_ES |
dc.identifier.issn | 2235-2988 | |
dc.identifier.uri | http://hdl.handle.net/10810/67473 | |
dc.description.abstract | Antibiotic resistance represents one of the greatest threats to global health. The spread of antibiotic resistance genes among bacteria occurs mostly through horizontal gene transfer via conjugation mediated by plasmids. This process implies a direct contact between a donor and a recipient bacterium which acquires the antibiotic resistance genes encoded by the plasmid and, concomitantly, the capacity to transfer the acquired plasmid to a new recipient. Classical assays for the measurement of plasmid transfer frequency (i.e., conjugation frequency) are often characterized by a high variability and, hence, they require many biological and technical replicates to reduce such variability and the accompanying uncertainty. In addition, classical conjugation assays are commonly tedious and time-consuming because they typically involve counting colonies on a large number of plates for the quantification of donors, recipients, and transconjugants (i.e., the bacteria that have received the genetic material by conjugation). Due to the magnitude of the antibiotic resistance problem, it is critical to develop reliable and rapid methods for the quantification of plasmid transfer frequency that allow the simultaneous analysis of many samples. Here, we present the development of a high-throughput, reliable, quick, easy, and cost-effective method to simultaneously accomplish and measure multiple conjugation events in 96-well plates, in which the quantification of donors, recipients, and transconjugants is estimated from the time required to reach a specific threshold value (OD600 value) in the bacterial growth curves. Our method successfully discriminates different plasmid transfer frequencies, yielding results that are equivalent to those obtained by a classical conjugation assay. | es_ES |
dc.description.sponsorship | The authors declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by MCIN/AEI/10.13039/501100011033 (PID2020-116495RB-I00) and the Basque Government (IT1578-22). AR-S received an UPV/EHU Training of pre-doctoral Research Staff Grant. NS-F received an Ikasiker grant from the Basque Government. | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | Frontiers Media | es_ES |
dc.relation | info:eu-repo/grantAgreement/MICINN/PID2020-116495RB-I00 | es_ES |
dc.rights | info:eu-repo/semantics/openAccess | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/es/ | * |
dc.subject | antibiotic resistance | es_ES |
dc.subject | bacterial conjugation | es_ES |
dc.subject | conjugation frequency | es_ES |
dc.subject | conjugation inhibitors | es_ES |
dc.subject | Escherichia coli | es_ES |
dc.subject | high-throughput screening platform | es_ES |
dc.subject | horizontal gene transfer | es_ES |
dc.title | Development of a high-throughput platform to measure plasmid transfer frequency | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.rights.holder | © 2023 Arbé-Carton, Rey-Sogo, Santos-Fernández, Altube, Garbisu, Arana and Alkorta. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. | es_ES |
dc.rights.holder | Atribución 3.0 España | * |
dc.relation.publisherversion | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1269732/full | es_ES |
dc.identifier.doi | 10.3389/fcimb.2023.1269732 | |
dc.departamentoes | Bioquímica y biología molecular | es_ES |
dc.departamentoes | Química aplicada | es_ES |
dc.departamentoeu | Biokimika eta biologia molekularra | es_ES |
dc.departamentoeu | Kimika aplikatua | es_ES |
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Except where otherwise noted, this item's license is described as © 2023 Arbé-Carton, Rey-Sogo, Santos-Fernández, Altube, Garbisu, Arana and Alkorta. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.