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Living Modified Organism
(LMO)
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Pseudomonas fluorescens strain SBW25 modified for biocontrol of fungal pathogens
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SBW25:tgl
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- Organization: Swedish University of Agricultural Sciences (SLU), Department of Microbiology () | BCH-CON-SCBD-49435-2Organization:Swedish University of Agricultural Sciences (SLU), Department of Microbiology ()Box 7025 750 07 Uppsala, Sweden,
,Phone:Fax:Email:Website:
The bacterium, Pseudomonas fluorescens, was chromosomally tagged with genes kilA, telAB conferring resistance to potassium tellurite, the constitutive promoter P-psbA, the gfp gene producing green fluorescent protein and luxAB genes encoding bioluminescence production. All these genes were inserted in order to monitor and trace the bacterium in the wheat plant-soil environment.
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The term “Recipient organism” refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas “Parental organisms” refers to those that were involved in cross breeding or cell fusion.
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BCH-ORGA-SCBD-48835-4 Organism Pseudomonas fluorescens (PSEFL)Bacteria
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pUTtgl
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- Electroporation
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
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BCH-GENE-SCBD-45846-5 Green Fluorescent Protein gene | Aequorea victoria (Crystal Jellyfish, Water Jellyfish, AEQVI)Protein coding sequence | Selectable marker genes and reporter genes
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BCH-GENE-SCBD-103753-1 Thylakoid membrane protein gene promoter | Amaranthus hybridus (Slim Amaranth, Green Amaranth, Pigweed)Promoter
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BCH-GENE-SCBD-103755-1 Luciferase alpha and beta subunit fusion gene | Vibrio harveyi (Vibrio, V. harveyi)Protein coding sequence | Selectable marker genes and reporter genes
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BCH-GENE-SCBD-103757-1 kilA gene | Enterobacter aerogenes (Enterobacter)Protein coding sequence | Selectable marker genes and reporter genes
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BCH-GENE-SCBD-103758-1 telAB gene | Enterobacter aerogenes (Enterobacter)Protein coding sequence | Selectable marker genes and reporter genes
Intended function of each constituent part of the insert in the GMO:
kilA, telAB: these genes confer resistance to the chemical compound potassium tellurite (K2TeO3). The use of these genes as marker genes was developed to suit strains that were aimed to be released in field trials, where it is not desirable to use antibiotic resistance genes as marker tools for tracking the bacteria. Makes selective plating on potassium tellurite possible.
P-psbA: a constitutive promoter.
GFP: GFP emits green light at 508 nm upon ultraviolet light illumination at 396 nm, no additional substrates needed. GFP is a measurement of the total number of cells since it is expressed regardless of metabolic activity in the cells.
LUXAB: The phenotype, bioluminescence, is dependent on the energy reserves of the cells, FMNH2. Therefore, the luxAB genes can be used to determine the number of metabolically active cells. Bioluminescence requires n-decanal as a substrate.
Information on the inserted DNA sequences
kilA, telAB: The tellurite resistance is encoded by the cryptic telAB and kilA genes of plasmid RK2 from Klebsiella aerogenes. PpsbA/gfp/luxAB: The PpsbA/gfp/luxAB genes were excised from plasmid pUTgfplux as a NotI fragment and ligated into the unique NotI-site of pUTtel.
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kilA, telAB: these genes confer resistance to the chemical compound potassium tellurite (K2TeO3). The use of these genes as marker genes was developed to suit strains that were aimed to be released in field trials, where it is not desirable to use antibiotic resistance genes as marker tools for tracking the bacteria. Makes selective plating on potassium tellurite possible.
P-psbA: a constitutive promoter.
GFP: GFP emits green light at 508 nm upon ultraviolet light illumination at 396 nm, no additional substrates needed. GFP is a measurement of the total number of cells since it is expressed regardless of metabolic activity in the cells.
LUXAB: The phenotype, bioluminescence, is dependent on the energy reserves of the cells, FMNH2. Therefore, the luxAB genes can be used to determine the number of metabolically active cells. Bioluminescence requires n-decanal as a substrate.
Information on the inserted DNA sequences
kilA, telAB: The tellurite resistance is encoded by the cryptic telAB and kilA genes of plasmid RK2 from Klebsiella aerogenes. PpsbA/gfp/luxAB: The PpsbA/gfp/luxAB genes were excised from plasmid pUTgfplux as a NotI fragment and ligated into the unique NotI-site of pUTtel.
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- Other (Biocontrol)
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