Construction of a Sinorhizobium meliloti strain carrying 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 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-P64...

Full description

Saved in:
Bibliographic Details
Published in:FEMS microbiology letters 2002-09, Vol.214 (2), p.165-170
Main Authors: Pistorio, M, Balague, L.J, Papa, M.F. del, Pich-Otero, A, Lodeiro, A, Hozbor, D.F, Lagares, A
Format: Article
Language:English
Subjects:
Alfalfa
Animal, plant and microbial ecology
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
Chromosomes
Chromosomes, Bacterial
Competitiveness
Fluorescence
Fundamental and applied biological sciences. Psychology
genes
Genetic Vectors
Genetics
Green fluorescent protein
Green Fluorescent Proteins
Labeling
Luminescent Proteins - genetics
Microbial ecology
Microbiology
Nitrogen Fixation
Nodulation
Nodules
Phenotype
Phenotypes
Proteins
RecA protein
Sinorhizobium meliloti
Sinorhizobium meliloti - genetics
Soil
Soil environment
Soil Microbiology
Symbiosis
tetracycline
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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.
ISSN:0378-1097
1574-6968
DOI:10.1111/j.1574-6968.2002.tb11341.x