Article Figures & Data
Figures
- FIG. 1.
Organ load in BALB/c mice infected i.n. with 1 × 104 CFU of B. pseudomallei K96243. •, lungs; ○, spleens. Symbols indicate counts per animal, and bars indicate means per group.
- FIG. 2.
(A) Attenuating transposon insertion sites in B. pseudomallei K96243. (B) Transposon insertion sites within the capsular polysaccharide biosynthesis region of B. pseudomallei K96243. Transposon insertion sites are indicated by vertical arrows. Genes and regions are not drawn to scale.
- FIG. 3.
Binding of monoclonal antibody 4V1H12 to the capsular polysaccharide of B. pseudomallei K96243 and selected transposon derivatives with insertions in the capsular polysaccharide coding region. (A) B. pseudomallei K96243 fluorescence view; (B) fluorescence view superimposed on phase-contrast image; (C and D) mutant 13H8 (wcbB); (E and F) mutant 1B9 (wcbC); (G and H) mutant 1A6 (wcbN).
- FIG. 4.
(A) Survival assay of mice infected with 1 × 103 CFU of B. pseudomallei K96243 (▪) or the wcbN mutant (▿). (B) Survival assay of mice infected with 1 × 103 of CFU B. pseudomallei K96243 (▪) or the wcbC mutant (▵).
- FIG. 5.
(A) Organ loads obtained from lungs of BALB/c mice infected i.n. with 1 × 103 CFU of wild-type (WT) B. pseudomallei K96243 (▴) or wcbC mutant strain 1B9 (B9) (▵). (B) Organ loads obtained from spleens of BALB/c mice infected i.n. with 1 × 103 CFU of B. pseudomallei K96243 (•) or wcbC mutant strain 1B9 (▵). Each symbol indicates counts from an individual mouse. Solid lines indicate mean values per group, and the broken lines show the limit of detection for this assay. NS, not statistically significant; S, significant (P < 0.05).
- FIG. 6.
Growth of B. pseudomallei K96243 and mutant strain 13B11 in M9 minimal medium with and without aromatic compound supplementation. ⧫, B. pseudomallei K96243 in M9 minimal medium; ▴, 13B11 in M9 minimal medium; □, B. pseudomallei K96243 in M9 minimal medium with aromatic compounds; ○, 13B11 in M9 minimal medium with aromatic compounds. The results indicate the means of three experiments each carried out in triplicate. Bars indicate standard deviations. O.D. 600, optical density at 600 nm.
- FIG. 7.
Protection assay of BALB/c mice vaccinated with B. pseudomallei mutant strain 13B11. ▪, naive mice; ⋄, mice vaccinated with 1 × 105 CFU of mutant strain 13B11; ▾, mice vaccinated with 1 × 106 CFU of mutant strain 13B11. Mice were challenged with 1 × 103 CFU of B. pseudomallei K96243 at day 35 postvaccination.
- FIG. 8.
(A) Organ loads obtained from lungs of BALB/c mice infected i.n. with 6 × 105 CFU of wild-type (WT) B. pseudomallei K96243 (•) or 7.5 × 105 CFU of aroB mutant strain 13B11 (▵). (B) Organ loads obtained from spleens of BALB/c mice infected i.n. with 6 × 105 CFU of wild-type B. pseudomallei K96243 (•) or 7.5 × 105 CFU of aroB mutant strain 13B11 (▵). Solid lines indicate mean values per group, and broken lines show the limit of detection for this assay. S, significant (P < 0.05).
Tables
- TABLE 1.
Transposon insertion sites identified in this studya
Mutant Disrupted gene Product Putative function Insertion distance from start codon (bp)b 7A6 wcbA Putative capsular polysaccharide export protein Capsular polysaccharide biosynthesis 1,301 13E8 wcbA Putative capsular polysaccharide export protein Capsular polysaccharide biosynthesis 713 13H8 wcbB Putative capsular polysaccharide glycosyltransferase biosynthesis protein Capsular polysaccharide biosynthesis 46 1B9 wcbC Putative capsular polysaccharide export protein Capsular polysaccharide biosynthesis 1,035 8F1 wcbC Putative capsular polysaccharide export protein Capsular polysaccharide biosynthesis 1,018 6E3 wcbC Putative capsular polysaccharide export protein Capsular polysaccharide biosynthesis 518 4A4 wcbD Putative ABC transporter transmembrane protein Capsular polysaccharide biosynthesis 961 JCX2C5 wzm Putative capsule polysaccharide export ABC transporter transmembrane protein Capsular polysaccharide biosynthesis 466 8C3 wcbE Putative glycosyltransferase Capsular polysaccharide biosynthesis 196 13G10 wcbE Putative glycosyltransferase Capsular polysaccharide biosynthesis 945 4H9 wcbE Putative glycosyltransferase Capsular polysaccharide biosynthesis 1,464 5E12 wcbE Putative glycosyltransferase Capsular polysaccharide biosynthesis 1,346 6G8 wcbH Putative glycosyltransferase Capsular polysaccharide biosynthesis 625 8H1 wcbH Putative glycosyltransferase Capsular polysaccharide biosynthesis 731 9G8 wcbH Putative glycosyltransferase Capsular polysaccharide biosynthesis 474 JCX2B10 wcbH Putative glycosyltransferase Capsular polysaccharide biosynthesis 75 4E6 wcbI Putative capsular polysaccharide biosynthesis protein Capsular polysaccharide biosynthesis 459 JCX2B2 wcbJ Putative capsular polysaccharide biosynthesis protein Capsular polysaccharide biosynthesis 516 1A6 wcbN Putative d-glycero-d-manno-heptose 1,7-bisphosphate phosphatase Capsular polysaccharide biosynthesis 258 13G3 wcbQ Putative capsular polysaccharide transmembrane export protein Capsular polysaccharide biosynthesis 711 12F5 wcbR Putative type I polyketide synthase Capsular polysaccharide biosynthesis 761 JCX2A9 wcbR Putative type I polyketide synthase Capsular polysaccharide biosynthesis 451 8A8 BPSL0776 RecA Homologous recombination, SOS response 207 7G7 BPSL2825 Putative PabB Biosynthesis of para-aminobenzoate 1,352 9D6 serC Phosphoserine aminotransferase Serine biosynthesis 360 6H5 trpG Anthranilate synthase component II Tryptophan biosynthesis 447 12C6 leuB Isopropylmalate dehydrogenase Leucine biosynthesis 769 8H10 leuB Isopropylmalate dehydrogenase Leucine biosynthesis 899 13B11 aroB Dehydroquinate synthase Chorismate biosynthesis 532 7A1 BPSL3147 Putative lipoprotein 39.16% amino acid identity to Shigella flexneri VacJ lipoprotein 618 6H4 BPSL1039 Putative ABC transporter Putative transmembrane ABC transporter 1,293 7G1 BPSL0634 Putative oxidoreductase Energy production and conversion 2,306 6A8 BPSL0634 Putative oxidoreductase Energy production and conversion 3,191 13D4 ppsA Putative phosphoenol pyruvate synthase Carbohydrate transport and metabolism 704 JCX2D8 Upstream of ihfA Putative integration host factor alpha subunit IhfA DNA replication, recombination, and repair NA 9F5 nth Putative endonuclease III DNA replication, recombination, and repair 150 7D6 nth Putative endonuclease III DNA replication, recombination, and repair 134 JCX2E12 BPSL0175/BPSL1060 Putative hypothetical bacteriophage protein Unknown 149 JCXD3 BPSL0175/BPSL1060 Putative hypothetical bacteriophage protein Unknown 123
