Development of molecular tools to monitor conjugative transfer in rhizobia

Autores
Torres Tejerizo, Gonzalo Arturo; Bañuelos, Luis Alfredo; Cervantes, Laura; Gaytán, Paul; Pistorio, Mariano; Romero, David; Brom, Susana
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Evolution of bacterial populations has been extensively driven by horizontal transfer events. Conjugative plasmid transfer is considered the principal contributor to gene exchange among bacteria. Several conjugative and mobilizable plasmids have been identified in rhizobia, and two major molecular mechanisms that regulate their transfer have been described, under laboratory conditions. The knowledge of rhizobial plasmid transfer regulation in natural environments is very poor. In this work we developed molecular tools to easily monitor the conjugative plasmid transfer in rhizobia by flow cytometry (FC) or microscopy. 24 cassettes were constructed by combining a variety of promotors, fluorescent proteins and antibiotic resistance genes, and used to tag plasmids and chromosome of donor strains. We were able to detect plasmid transfer after conversion of non-fluorescent recipients into fluorescent transconjugants. Flow cytometry (FC) was optimized to count donor, recipient and transconjugant strains to determine conjugative transfer frequencies. Results were similar, when determined either by FC or by viable counts. Our constructions also allowed the visualization of transconjugants in crosses performed on bean roots. The tools presented here may also be used for other purposes, such as analysis of transcriptional fusions or single-cell tagging. Application of the system will allow the survey of how different environmental conditions or other regulators modulate plasmid transfer in rhizobia.
Fil: Torres Tejerizo, Gonzalo Arturo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentina
Fil: Bañuelos, Luis Alfredo. Universidad Nacional Autónoma de México; México
Fil: Cervantes, Laura. Universidad Nacional Autónoma de México; México
Fil: Gaytán, Paul. Universidad Nacional Autónoma de México; México
Fil: Pistorio, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentina
Fil: Romero, David. Universidad Nacional Autónoma de México; México
Fil: Brom, Susana. Universidad Nacional Autónoma de México; México
Materia
CONJUGATIVE TRANSFER
FLUORESCENT CASSETTES
PLASMID
RHIZOBIA
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/99884
Acceder
id CONICETDig_621737895a255ea51b8d831e02a246fa
oai_identifier_str oai:ri.conicet.gov.ar:11336/99884
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Development of molecular tools to monitor conjugative transfer in rhizobiaTorres Tejerizo, Gonzalo ArturoBañuelos, Luis AlfredoCervantes, LauraGaytán, PaulPistorio, MarianoRomero, DavidBrom, SusanaCONJUGATIVE TRANSFERFLUORESCENT CASSETTESPLASMIDRHIZOBIAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Evolution of bacterial populations has been extensively driven by horizontal transfer events. Conjugative plasmid transfer is considered the principal contributor to gene exchange among bacteria. Several conjugative and mobilizable plasmids have been identified in rhizobia, and two major molecular mechanisms that regulate their transfer have been described, under laboratory conditions. The knowledge of rhizobial plasmid transfer regulation in natural environments is very poor. In this work we developed molecular tools to easily monitor the conjugative plasmid transfer in rhizobia by flow cytometry (FC) or microscopy. 24 cassettes were constructed by combining a variety of promotors, fluorescent proteins and antibiotic resistance genes, and used to tag plasmids and chromosome of donor strains. We were able to detect plasmid transfer after conversion of non-fluorescent recipients into fluorescent transconjugants. Flow cytometry (FC) was optimized to count donor, recipient and transconjugant strains to determine conjugative transfer frequencies. Results were similar, when determined either by FC or by viable counts. Our constructions also allowed the visualization of transconjugants in crosses performed on bean roots. The tools presented here may also be used for other purposes, such as analysis of transcriptional fusions or single-cell tagging. Application of the system will allow the survey of how different environmental conditions or other regulators modulate plasmid transfer in rhizobia.Fil: Torres Tejerizo, Gonzalo Arturo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Bañuelos, Luis Alfredo. Universidad Nacional Autónoma de México; MéxicoFil: Cervantes, Laura. Universidad Nacional Autónoma de México; MéxicoFil: Gaytán, Paul. Universidad Nacional Autónoma de México; MéxicoFil: Pistorio, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; ArgentinaFil: Romero, David. Universidad Nacional Autónoma de México; MéxicoFil: Brom, Susana. Universidad Nacional Autónoma de México; MéxicoElsevier Science2015-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/99884Torres Tejerizo, Gonzalo Arturo; Bañuelos, Luis Alfredo; Cervantes, Laura; Gaytán, Paul; Pistorio, Mariano; et al.; Development of molecular tools to monitor conjugative transfer in rhizobia; Elsevier Science; Journal of Microbiological Methods; 117; 10-2015; 155-1630167-7012CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0167701215300385info:eu-repo/semantics/altIdentifier/doi/10.1016/j.mimet.2015.08.005info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:26:06Zoai:ri.conicet.gov.ar:11336/99884instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 11:26:06.312CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Development of molecular tools to monitor conjugative transfer in rhizobia
title Development of molecular tools to monitor conjugative transfer in rhizobia
spellingShingle Development of molecular tools to monitor conjugative transfer in rhizobia
Torres Tejerizo, Gonzalo Arturo
CONJUGATIVE TRANSFER
FLUORESCENT CASSETTES
PLASMID
RHIZOBIA
title_short Development of molecular tools to monitor conjugative transfer in rhizobia
title_full Development of molecular tools to monitor conjugative transfer in rhizobia
title_fullStr Development of molecular tools to monitor conjugative transfer in rhizobia
title_full_unstemmed Development of molecular tools to monitor conjugative transfer in rhizobia
title_sort Development of molecular tools to monitor conjugative transfer in rhizobia
dc.creator.none.fl_str_mv Torres Tejerizo, Gonzalo Arturo
Bañuelos, Luis Alfredo
Cervantes, Laura
Gaytán, Paul
Pistorio, Mariano
Romero, David
Brom, Susana
author Torres Tejerizo, Gonzalo Arturo
author_facet Torres Tejerizo, Gonzalo Arturo
Bañuelos, Luis Alfredo
Cervantes, Laura
Gaytán, Paul
Pistorio, Mariano
Romero, David
Brom, Susana
author_role author
author2 Bañuelos, Luis Alfredo
Cervantes, Laura
Gaytán, Paul
Pistorio, Mariano
Romero, David
Brom, Susana
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv CONJUGATIVE TRANSFER
FLUORESCENT CASSETTES
PLASMID
RHIZOBIA
topic CONJUGATIVE TRANSFER
FLUORESCENT CASSETTES
PLASMID
RHIZOBIA
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Evolution of bacterial populations has been extensively driven by horizontal transfer events. Conjugative plasmid transfer is considered the principal contributor to gene exchange among bacteria. Several conjugative and mobilizable plasmids have been identified in rhizobia, and two major molecular mechanisms that regulate their transfer have been described, under laboratory conditions. The knowledge of rhizobial plasmid transfer regulation in natural environments is very poor. In this work we developed molecular tools to easily monitor the conjugative plasmid transfer in rhizobia by flow cytometry (FC) or microscopy. 24 cassettes were constructed by combining a variety of promotors, fluorescent proteins and antibiotic resistance genes, and used to tag plasmids and chromosome of donor strains. We were able to detect plasmid transfer after conversion of non-fluorescent recipients into fluorescent transconjugants. Flow cytometry (FC) was optimized to count donor, recipient and transconjugant strains to determine conjugative transfer frequencies. Results were similar, when determined either by FC or by viable counts. Our constructions also allowed the visualization of transconjugants in crosses performed on bean roots. The tools presented here may also be used for other purposes, such as analysis of transcriptional fusions or single-cell tagging. Application of the system will allow the survey of how different environmental conditions or other regulators modulate plasmid transfer in rhizobia.
Fil: Torres Tejerizo, Gonzalo Arturo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentina
Fil: Bañuelos, Luis Alfredo. Universidad Nacional Autónoma de México; México
Fil: Cervantes, Laura. Universidad Nacional Autónoma de México; México
Fil: Gaytán, Paul. Universidad Nacional Autónoma de México; México
Fil: Pistorio, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentina
Fil: Romero, David. Universidad Nacional Autónoma de México; México
Fil: Brom, Susana. Universidad Nacional Autónoma de México; México
description Evolution of bacterial populations has been extensively driven by horizontal transfer events. Conjugative plasmid transfer is considered the principal contributor to gene exchange among bacteria. Several conjugative and mobilizable plasmids have been identified in rhizobia, and two major molecular mechanisms that regulate their transfer have been described, under laboratory conditions. The knowledge of rhizobial plasmid transfer regulation in natural environments is very poor. In this work we developed molecular tools to easily monitor the conjugative plasmid transfer in rhizobia by flow cytometry (FC) or microscopy. 24 cassettes were constructed by combining a variety of promotors, fluorescent proteins and antibiotic resistance genes, and used to tag plasmids and chromosome of donor strains. We were able to detect plasmid transfer after conversion of non-fluorescent recipients into fluorescent transconjugants. Flow cytometry (FC) was optimized to count donor, recipient and transconjugant strains to determine conjugative transfer frequencies. Results were similar, when determined either by FC or by viable counts. Our constructions also allowed the visualization of transconjugants in crosses performed on bean roots. The tools presented here may also be used for other purposes, such as analysis of transcriptional fusions or single-cell tagging. Application of the system will allow the survey of how different environmental conditions or other regulators modulate plasmid transfer in rhizobia.
publishDate 2015
dc.date.none.fl_str_mv 2015-10
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/99884
Torres Tejerizo, Gonzalo Arturo; Bañuelos, Luis Alfredo; Cervantes, Laura; Gaytán, Paul; Pistorio, Mariano; et al.; Development of molecular tools to monitor conjugative transfer in rhizobia; Elsevier Science; Journal of Microbiological Methods; 117; 10-2015; 155-163
0167-7012
CONICET Digital
CONICET
url http://hdl.handle.net/11336/99884
identifier_str_mv Torres Tejerizo, Gonzalo Arturo; Bañuelos, Luis Alfredo; Cervantes, Laura; Gaytán, Paul; Pistorio, Mariano; et al.; Development of molecular tools to monitor conjugative transfer in rhizobia; Elsevier Science; Journal of Microbiological Methods; 117; 10-2015; 155-163
0167-7012
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0167701215300385
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.mimet.2015.08.005
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.982451

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