Article Figures & Data
Figures
- Fig. 1.
Deduced amino acid sequence of Tn5-flanking DNA from theB. abortus rough mutants aligned with amino acid sequences from GenBank. (A) Alignment of CA180 ORF product withRhizobium sp. strain NoeI and Vibrio choleraePMM; (B) alignment of CA353 ORF product with V. choleraeRfbD-like gene product and V. cholerae oxidoreductase (oxido.); (C) alignment of CA533 ORF product with V. cholerae RfbV and an outer membrane protein (OMP) precursor ofV. cholerae; (D) alignment of CA613 ORF product with RfbA ofP. aeruginosa and TrsF of Y. enterocolitica. Conserved amino acids are indicated by vertical lines, and similar amino acids are indicated by colons. Numbers at the beginnings and ends of sequences indicate the positions of published amino acids. Dashes indicate spaces inserted to improve alignment. Alignments were performed with CLUSTAL V. See the text for complete descriptions of the genes.
- Fig. 2.
Immunoblot of whole-cell lysates from B. abortus smooth and rough strains electrophoresed on a 12% (wt/vol) gel, transferred to a polyvinylidene difluoride membrane, and reacted with anti-O-antigen antibody (MAb 39). Lane 1, S2308; lane 2, S19; lane 3, RB51; lane 4, CA613; lane 5, CA533; lane 6, CA353; lane 7, CA180. Positions of molecular mass markers (in kilodaltons) are shown on the left.
- Fig. 3.
Immunoblot of whole-cell lysates from B. abortus smooth and rough strains electrophoresed on a 12% (wt/vol) gel, transferred to a polyvinylidene difluoride membrane, hydrolyzed in 10% acetic acid, and reacted with anti-lipid A antibody (MAb 177). Lane 1, molecular mass markers (sizes in kilodaltons are shown on the left); lane 2, S2308; lane 3, S19; lane 4, RB51; lane 5, CA613; lane 6, CA533; lane 7, CA353; lane 8, CA180.
- Fig. 4.
Complement-mediated killing of B. abortusstrains in bovine sera. Complement-mediated killing was performed as described in Materials and Methods. Bacterial counts are reported as log CFU/ml (fresh sera) − log CFU/ml (heat-inactivated sera). The results represent means from four independent assays (P< 0.0001).
- Fig. 5.
Percent survival of B. abortus strains in bovine peripheral blood macrophages. Macrophage-mediated killing was performed as described in Materials and Methods. Results are expressed as percent survival in comparison to that of S2308 and represent the averages from three independent trials (P < 0.05).
- Fig. 6.
Bactericidal effect of PmB on B. abortusstrains. PmB-mediated killing was performed as described in Materials and Methods. Average results of three assays are expressed as percentages of the brucellae surviving in wells incubated in the absence of PmB. Asterisks indicate statistically significant results as noted in the text. Symbols: ▪, S2308; •, RB51; ▴, CA180; ⧫, CA353; □, CA533; ○, CA613.
Tables
- Table 1.
Bacterial counts in mouse spleens examined 1, 2, and 6 weeks after i.p. infection
Strain CFU/spleena at wk: 1 2 6 Wild type (S2308) 4.88 × 106 (2.83 × 105–3.06 × 107) 2.34 × 106 (6.45 × 105–4.45 × 106) 1.34 × 106 (1.12 × 106–3.69 × 106) Mutants CA180 0 (0–1.06 × 102) 0 (0–4.93 × 102) 0 (0) CA353 1.94 × 103 (1.23 × 101–3.51 × 104) 8.4 × 101 (0–5.59 × 103) 0 (0) CA533 1.19 × 104 (2.20 × 102–7.28 × 104) 5.51 × 102 (2.50 × 102–4.27 × 104) 0 (0–1.62 × 104) CA613 7.52 × 102 (2.17 × 102–3.20 × 103) 5.51 × 102 (0–7.38 × 103) 0 (0–1.12 × 102) ↵a Bacterial counts in the mouse spleens are represented as medians, with the lower (25%) and upper (75%) quartiles presented in parentheses.
