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

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&...

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Autores principales: Pistorio, Mariano, Balagué, Laura Josefina, Del Papa, María Florencia, Pich Otero, Augusto J. L., Lodeiro, Aníbal Roberto, Hozbor, Daniela Flavia, Lagares, Antonio
Formato: Articulo
Lenguaje:Inglés
Publicado: 2002
Materias:
Ciencias Exactas
Green fluorescent protein
Nodulation
Sinorhizobium meliloti
Soil
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/84925
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-84925
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
Green fluorescent protein
Nodulation
Sinorhizobium meliloti
Soil
spellingShingle Ciencias Exactas
Green fluorescent protein
Nodulation
Sinorhizobium meliloti
Soil
Pistorio, Mariano
Balagué, Laura Josefina
Del Papa, María Florencia
Pich Otero, Augusto J. L.
Lodeiro, Aníbal Roberto
Hozbor, Daniela Flavia
Lagares, Antonio
Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T
topic_facet Ciencias Exactas
Green fluorescent protein
Nodulation
Sinorhizobium meliloti
Soil
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.
format Articulo
Articulo
author Pistorio, Mariano
Balagué, Laura Josefina
Del Papa, María Florencia
Pich Otero, Augusto J. L.
Lodeiro, Aníbal Roberto
Hozbor, Daniela Flavia
Lagares, Antonio
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_sort Pistorio, Mariano
title Construction of a Sinorhizobium meliloti strain carrying a stable and non-transmissible chromosomal single copy of the green fluorescent protein GFP-P64L/S65T
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
publishDate 2002
url http://sedici.unlp.edu.ar/handle/10915/84925
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