Abstract
Agrobacterium tumefaciens is the main plant biotechnology gene transfer tool with host range which can be extended to non-plant eukaryotic organisms under laboratory conditions. Known medical cases of Agrobacterium species isolation from bloodstream infections necessitate the assessment of biosafety-related risks of A. tumefaciens encounters with mammalian organisms. Here, we studied the survival of A. tumefaciens in bloodstream of mice injected with bacterial cultures. Bacterial titers of 108 CFU were detected in the blood of the injected animals up to two weeks after intravenous injection. Agrobacteria carrying Cauliflower mosaic virus (CaMV) 35S promoter-based constructs and isolated from the injected mice retained their capacity to promote green fluorescent protein (GFP) synthesis in Nicotiana benthamiana leaves. To examine whether or not the injected agrobacteria are able to express in mouse organs, we used an intron-containing GFP (GFPi) reporter driven either by a cytomegalovirus (CMV) promoter or by a CaMV 35S promoter. Western and northern blot analyses as well as RT-PCR analysis of liver, spleen and lung of mice injected with A. tumefaciens detected neither GFP protein nor its transcripts. Thus, bacteraemia induced in mice by A. tumefaciens does not lead to detectible levels of genetic transformation of mouse organs.
Highlights
Engineered plants often represent a preferred source of recombinant proteins and biopharmaceuticals for human consumption [1,2]
To examine whether or not the injected agrobacteria are able to express in mouse organs, we used an intron-containing green fluorescent protein (GFP) (GFPi) reporter driven either by a cytomegalovirus (CMV) promoter or by a Cauliflower mosaic virus (CaMV) 35S promoter
Bacteraemia induced in mice by A. tumefaciens does not lead to detectible levels of genetic transformation of mouse organs
Summary
Engineered plants often represent a preferred source of recombinant proteins and biopharmaceuticals for human consumption [1,2]. Genetic transformation of plants is usually achieved using Agrobacterium tumefaciens. This bacterium is a soil-borne, nonpathogenic for humans microorganism which can transfer its T-DNA into the genomes of plants and of human cultured cells (for review, see [3,4,5]). 17] require the assessment of biosafety-related implications of Agrobacterium invasion of mammalian organisms. We studied whether or not intravenously injected A .tumefaciens can survive in mouse bloodstream and direct expression of its TDNA within mouse organs. Agrobacterium persisted in the bloodstream for up to two weeks post injection, it failed to express the reporter GFP gene in such diverse organs as spleen, liver and lung. Agrobacterium induces bacteremia in mice, but does not cause detectible genetic alteration of mouse tissues
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