title: Characterization of the Burkholderia pseudomallei K96243 capsular polysaccharide I coding region.
abstract: Burkholderia pseudomallei is the causative agent of melioidosis, a disease endemic to regions of Southeast Asia and Northern Australia. Both humans and a range of other animal species are susceptible to melioidosis, and the production of a group 3 polysaccharide capsule in B. pseudomallei is essential for virulence. B. pseudomallei capsular polysaccharide (CPS) I comprises unbranched manno-heptopyranose residues and is encoded by a 34.5-kb locus on chromosome 1. Despite the importance of this locus, the role of all of the genes within this region is unclear. We inactivated 18 of these genes and analyzed their phenotype using Western blotting and immunofluorescence staining. Furthermore, by combining this approach with bioinformatic analysis, we were able to develop a model for CPS I biosynthesis and export. We report that inactivating gmhA, wcbJ, and wcbN in B. pseudomallei K96243 retains the immunogenic integrity of the polysaccharide despite causing attenuation in the BALB/c murine infection model. Mice immunized with the B. pseudomallei K96243 mutants lacking a functional copy of either gmhA or wcbJ were afforded significant levels of protection against a wild-type B. pseudomallei K96243 challenge.
authors.affiliation: Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom.
authors.forename: Jon
authors.initials: J
authors.lastname: Cuccui
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Milne
authors.forename: Nicholas
authors.initials: N
authors.lastname: Harmer
authors.forename: Alison J
authors.initials: AJ
authors.lastname: George
authors.forename: Sarah V
authors.initials: SV
authors.lastname: Harding
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Dean
authors.forename: Andrew E
authors.initials: AE
authors.lastname: Scott
authors.forename: Mitali
authors.initials: M
authors.lastname: Sarkar-Tyson
authors.forename: Brendan W
authors.initials: BW
authors.lastname: Wren
authors.forename: Richard W
authors.initials: RW
authors.lastname: Titball
authors.forename: Joann L
authors.initials: JL
authors.lastname: Prior
mesh_terms: D000818:Animals; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D053898:Biosynthetic Pathways / Q000235:genetics*; D015153:Blotting, Western; D016957:Burkholderia pseudomallei / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D005260:Female; D005455:Fluorescent Antibody Technique; D055786:Gene Knockout Techniques; D056426:Genetic Loci; D008554:Melioidosis / Q000382:microbiology / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D005810:Multigene Family; D011135:Polysaccharides, Bacterial / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014774:Virulence; D037521:Virulence Factors / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D001428:Bacterial Vaccines; D011135:Polysaccharides, Bacterial; D014613:Vaccines, Attenuated; D037521:Virulence Factors