Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T

Autores
Pistorio, Mariano; Balagué, Laura Josefina; Del Papa, María Florencia; Pich Otero, Augusto J. L.; Lodeiro, Aníbal Roberto; Hozbor, Daniela Flavia; Lagares, Antonio
Año de publicación
2002
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
A single copy of the green fluorescent protein (GFP)-encoding gene gfp-P64L/S65T under the control of the constitutive nptII promoter was introduced in a neutral region of the Sinorhizobium meliloti chromosome, between the genes recA and alaS. Within the same chromosomal region downstream of gfp-P64L/S65T a tetracycline (Tc) resistant cassette was also inserted. Both markers were very stable during at least 40 bacterial generations without any selective pressure. Similarly, the gfp-Tc cassette was stable and functional in all rhizobia that were recovered from alfalfa nodules. The GFP-associated fluorescence derived from the (single copy) chromosomal gfp-P64L/S65T allowed detection of rhizobia during the colonisation of the root, infection thread formation, and nodule development. The gfp-Tc rhizobia showed indistinguishable phenotypes for nodulation, competitiveness, and nitrogen-fixation from the parental strain. The labelling system described here can be used for the stable fluorescent tagging of S. meliloti strains allowing their detection in biologically complex soil environments.
Facultad de Ciencias Exactas
Instituto de Biotecnologia y Biologia Molecular
Materia
Ciencias Exactas
Green fluorescent protein
Nodulation
Sinorhizobium meliloti
Soil
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/84925
Acceder
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oai_identifier_str oai:sedici.unlp.edu.ar:10915/84925
network_acronym_str SEDICI
repository_id_str 1329
network_name_str SEDICI (UNLP)
spelling Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65TPistorio, MarianoBalagué, Laura JosefinaDel Papa, María FlorenciaPich Otero, Augusto J. L.Lodeiro, Aníbal RobertoHozbor, Daniela FlaviaLagares, AntonioCiencias ExactasGreen fluorescent proteinNodulationSinorhizobium melilotiSoilA single copy of the green fluorescent protein (GFP)-encoding gene <i>gfp</i>-P64L/S65T under the control of the constitutive <i>nptII</i> promoter was introduced in a neutral region of the <i>Sinorhizobium meliloti</i> chromosome, between the genes <i>recA</i> and <i>alaS</i>. Within the same chromosomal region downstream of <i>gfp</i>-P64L/S65T a tetracycline (Tc) resistant cassette was also inserted. Both markers were very stable during at least 40 bacterial generations without any selective pressure. Similarly, the <i>gfp</i>-Tc cassette was stable and functional in all rhizobia that were recovered from alfalfa nodules. The GFP-associated fluorescence derived from the (single copy) chromosomal <i>gfp</i>-P64L/S65T allowed detection of rhizobia during the colonisation of the root, infection thread formation, and nodule development. The <i>gfp</i>-Tc rhizobia showed indistinguishable phenotypes for nodulation, competitiveness, and nitrogen-fixation from the parental strain. The labelling system described here can be used for the stable fluorescent tagging of <i>S. meliloti</i> strains allowing their detection in biologically complex soil environments.Facultad de Ciencias ExactasInstituto de Biotecnologia y Biologia Molecular2002info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf165-170http://sedici.unlp.edu.ar/handle/10915/84925enginfo:eu-repo/semantics/altIdentifier/issn/0378-1097info:eu-repo/semantics/altIdentifier/doi/10.1016/S0378-1097(02)00812-1info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-29T11:16:15Zoai:sedici.unlp.edu.ar:10915/84925Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-29 11:16:16.25SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T
title Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T
spellingShingle Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T
Pistorio, Mariano
Ciencias Exactas
Green fluorescent protein
Nodulation
Sinorhizobium meliloti
Soil
title_short Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T
title_full Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T
title_fullStr Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T
title_full_unstemmed Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T
title_sort Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T
dc.creator.none.fl_str_mv Pistorio, Mariano
Balagué, Laura Josefina
Del Papa, María Florencia
Pich Otero, Augusto J. L.
Lodeiro, Aníbal Roberto
Hozbor, Daniela Flavia
Lagares, Antonio
author Pistorio, Mariano
author_facet Pistorio, Mariano
Balagué, Laura Josefina
Del Papa, María Florencia
Pich Otero, Augusto J. L.
Lodeiro, Aníbal Roberto
Hozbor, Daniela Flavia
Lagares, Antonio
author_role author
author2 Balagué, Laura Josefina
Del Papa, María Florencia
Pich Otero, Augusto J. L.
Lodeiro, Aníbal Roberto
Hozbor, Daniela Flavia
Lagares, Antonio
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Ciencias Exactas
Green fluorescent protein
Nodulation
Sinorhizobium meliloti
Soil
topic Ciencias Exactas
Green fluorescent protein
Nodulation
Sinorhizobium meliloti
Soil
dc.description.none.fl_txt_mv A single copy of the green fluorescent protein (GFP)-encoding gene <i>gfp</i>-P64L/S65T under the control of the constitutive <i>nptII</i> promoter was introduced in a neutral region of the <i>Sinorhizobium meliloti</i> chromosome, between the genes <i>recA</i> and <i>alaS</i>. Within the same chromosomal region downstream of <i>gfp</i>-P64L/S65T a tetracycline (Tc) resistant cassette was also inserted. Both markers were very stable during at least 40 bacterial generations without any selective pressure. Similarly, the <i>gfp</i>-Tc cassette was stable and functional in all rhizobia that were recovered from alfalfa nodules. The GFP-associated fluorescence derived from the (single copy) chromosomal <i>gfp</i>-P64L/S65T allowed detection of rhizobia during the colonisation of the root, infection thread formation, and nodule development. The <i>gfp</i>-Tc rhizobia showed indistinguishable phenotypes for nodulation, competitiveness, and nitrogen-fixation from the parental strain. The labelling system described here can be used for the stable fluorescent tagging of <i>S. meliloti</i> strains allowing their detection in biologically complex soil environments.
Facultad de Ciencias Exactas
Instituto de Biotecnologia y Biologia Molecular
description A single copy of the green fluorescent protein (GFP)-encoding gene <i>gfp</i>-P64L/S65T under the control of the constitutive <i>nptII</i> promoter was introduced in a neutral region of the <i>Sinorhizobium meliloti</i> chromosome, between the genes <i>recA</i> and <i>alaS</i>. Within the same chromosomal region downstream of <i>gfp</i>-P64L/S65T a tetracycline (Tc) resistant cassette was also inserted. Both markers were very stable during at least 40 bacterial generations without any selective pressure. Similarly, the <i>gfp</i>-Tc cassette was stable and functional in all rhizobia that were recovered from alfalfa nodules. The GFP-associated fluorescence derived from the (single copy) chromosomal <i>gfp</i>-P64L/S65T allowed detection of rhizobia during the colonisation of the root, infection thread formation, and nodule development. The <i>gfp</i>-Tc rhizobia showed indistinguishable phenotypes for nodulation, competitiveness, and nitrogen-fixation from the parental strain. The labelling system described here can be used for the stable fluorescent tagging of <i>S. meliloti</i> strains allowing their detection in biologically complex soil environments.
publishDate 2002
dc.date.none.fl_str_mv 2002
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
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://sedici.unlp.edu.ar/handle/10915/84925
url http://sedici.unlp.edu.ar/handle/10915/84925
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/0378-1097
info:eu-repo/semantics/altIdentifier/doi/10.1016/S0378-1097(02)00812-1
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
165-170
dc.source.none.fl_str_mv reponame:SEDICI (UNLP)
instname:Universidad Nacional de La Plata
instacron:UNLP
reponame_str SEDICI (UNLP)
collection SEDICI (UNLP)
instname_str Universidad Nacional de La Plata
instacron_str UNLP
institution UNLP
repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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score 13.070432

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