FIG 5.

(A) Mutants of the wbiA, wbiD, oacA, and wcbB genes exhibited PCR products of corresponding deletion alleles. Lane M represents a 100-bp ladder marker. Three LPS mutants were constructed with the K96243 wild type (lane WT), the mutant defective in wbiA (BPSL2680) (lane ΔwbiA), the mutant defective in wbiD (BPSL2677) (lane ΔwbiD), and the mutant defective in (BPSL1936) (lane ΔoacA). Lane ΔwbiA complement contains a complemented strain of the wbiA mutant. Two capsule mutants were constructed with the 4095a (NM) wild type and the 4095c (M) wild type. Lanes ΔwcbB contain a mutant defective in ΔwcbB (BPSL2808). (B) SDS-PAGE and a silver stain of LPS extracts of the wbiA, oacA, and wcbB mutants but not the wbiD mutant exhibited an LPS ladder pattern identical to that of the wild types. Lane M contains standard protein markers. Numbers at the left are molecular sizes (in base pairs). (C) Western blotting of LPS extracts of the wbiA, wbiD, oacA, and wcbB mutants probed with OPS-specific MAb 9D5 demonstrated a B. pseudomallei mutant without OPS (ΔwbiD) and the 2-O-acetylation moiety (ΔwbiA) and isolates of 4095c that did not react with MAb. Numbers at the left are molecular sizes (in base pairs). (D) Western blotting of LPS extracts of wbiA, wbiD, oacA, and wcbB mutants probed with capsule-specific MAb 4B11 demonstrated that only wcbB capsule mutants (ΔwcbB) did not react with capsule-specific MAb 4B11. Numbers at the left are molecular sizes (in base pairs).