agrobacterium tumefaciens-induced bacteraemia does not lead to reporter gene expression in mouse organs农杆菌属tumefaciens-induced菌血症不会导致记者基因表达在小鼠器官.pdfVIP

Agrobacterium tumefaciens-Induced Bacteraemia Does Not Lead to Reporter Gene Expression in Mouse Organs 1 1 1 3 2 Igor V. Petrunia , Olga Y. Frolova , Tatiana V. Komarova , Sergey L. Kiselev , Vitaly Citovsky , Yuri L. Dorokhov1,3* 1 A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia, 2 Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, New York, United States of America, 3 N.I. Vavilov Institute of General Genetics, Russian Academy of Science, Moscow, Russia 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 wi