Characterization of two bioluminescent Rhizobium meliloti strains constructed for field releases

The deliberate release of genetically engineered microorganisms requires a thorough characterization of the microbes in question. For the two bioluminescent Rhizobium meliloti strains, L1 and L33 [Selbitschka et al. (1992) Mol Ecol 1: 9-19; Selbitschka et al. (1995) FEMS Microbiol Ecol 16: 223-232],...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 1996-05, Vol.45 (4), p.509-512
Main Authors: Dammann-Kalinowski, T, Niemann, S, Keller, M, Selbitschka, W, Tebbe, C.C, Puhler, A
Format: Article
Language:English
Subjects:
Bacterial Proteins - biosynthesis
Bacterial Proteins - genetics
Base Sequence
Biological and medical sciences
Biology of microorganisms of confirmed or potential industrial interest
Biotechnology
Containment of Biohazards
Environment and pollution
Environmental Microbiology
Fundamental and applied biological sciences. Psychology
Genes, Synthetic
Genetically engineered organisms behavior (microorganisms, plants, animals)
Genetics
Industrial applications and implications. Economical aspects
Luciferases - biosynthesis
Luciferases - genetics
Luminescent Measurements
Medicago sativa - microbiology
Mission oriented research
Molecular Sequence Data
Phenotype
plant nutrition
Recombinant Fusion Proteins - biosynthesis
Rhizobium meliloti
Sinorhizobium meliloti - genetics
Species Specificity
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Summary:The deliberate release of genetically engineered microorganisms requires a thorough characterization of the microbes in question. For the two bioluminescent Rhizobium meliloti strains, L1 and L33 [Selbitschka et al. (1992) Mol Ecol 1: 9-19; Selbitschka et al. (1995) FEMS Microbiol Ecol 16: 223-232], designated for field release, the sites of genetic modifications in the chromosomes were sequenced from amplified genomic DNA. This indicated no unexpected alterations in the nucleotide sequence. The bioluminescent phenotype was stably inherited over more than 100 generations in liquid cultures. The presence of the luciferase gene in both strains did not have secondary effects on a variety of metabolic pathways as assessed by the Biology GN system. A specific polymerase chain reaction amplification, based on the chromosomal insertion site of the luc cassette, allowed the discrimination between the two strains and thus simplifies monitoring. The RecA-deficient strain L1 showed a strongly (more than 90%) reduced ability to nodulate alfalfa in competition with its parent strain R. meliloti 2011 and its RecA+ counterpart L33.
ISSN:0175-7598
1432-0614
DOI:10.1007/s002530050720