Characterisation of an acapsular mutant of Burkholderia pseudomallei identified by signature tagged mutagenesis


Atkins, T; Prior, R; Mack, K; Russell, P; Nelson, M; Prior, J; Ellis, J; Oyston, PCF; Dougan, G; Titball, RW; (2002) Characterisation of an acapsular mutant of Burkholderia pseudomallei identified by signature tagged mutagenesis. Journal of medical microbiology, 51 (7). pp. 539-47. ISSN 0022-2615

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Abstract

A Burkholderia pseudomallei mutant which was attenuated in a mouse model of melioidosis was identified by a signature tagged mutagenesis approach. The transposon was shown to be inserted into a gene within the capsular biosynthetic operon. Compared with the wild-type bacteria this mutant demonstrated a 10(5)-fold increase in the median lethal dose in a mouse model and it did not react with a monoclonal antibody against high mol. wt polysaccharide of B. pseudomallei. To determine the kinetics of infection, mice were dosed intraperitoneally (i.p.) and intravenously (i.v.) with mutant and wild-type bacteria. After i.p challenge, the number of mutant bacteria in the peritoneal cavity declined, whereas wild-type bacteria proliferated. When administered by the i.v. route, the mutant was able to cause disease but the time to death was increased compared with the wild type. Mice were dosed with the mutant and subsequently challenged with wild-type B. pseudomallei, but the mutant failed to induce a protective immune response.

Item Type: Article
Keywords: Bacterial virulence genes, pseudomonas-pseudomallei, transposon, mutagenesis, subtractive hybridization, intracellular survival, negative selection, tn5 insertion, murine model, melioidosis, polysaccharide, Animal, Antibodies, Monoclonal, immunology, Bacterial Capsules, genetics, immunology, Burkholderia pseudomallei, genetics, immunology, pathogenicity, Disease Models, Animal, Female, Immunoblotting, Injections, Intraperitoneal, Injections, Intravenous, Kinetics, Melioidosis, microbiology, Mice, Mice, Inbred BALB C, Mutagenesis, Mutation, Virulence, genetics, immunology
Faculty and Department: Faculty of Infectious and Tropical Diseases > Dept of Pathogen Molecular Biology
PubMed ID: 12132769
Web of Science ID: 176774600002
URI: http://researchonline.lshtm.ac.uk/id/eprint/17731

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