Red indicates homology of 78-100% and blue indicates an inversion region with equal homology Serum sensitivity Previous studies have shown that B. pseudomallei strains with type B2 or rough type O-antigens display an increased sensitivity to killing by 30% NHS [11, 23]. To determine if near-neighbors showed the same effect, eleven diverse Burkholderia strains expressing type A, B, or B2 O-antigen were assayed for serum sensitivity. All
type A strains, B. thailandensis E264, MSMB59, MSMB60, and B. oklahomensis E0147 showed a slight resistance to serum killing, except B. thailandensis TXDOH which was sensitive to serum killing. The type B2 B. thailandensis 82172 showed almost no difference in growth, and all other strains were sensitive to killing by 30% NHS, most notably B. ubonensis MSMB108 (Figure Ralimetinib 3). Figure 3 Serum sensitivity of B. pseudomallei near-neighbors. B. thailandensis E264, MSMB59, MSMB60 and B. oklahomensis E0147 showed a slight resistance to killing by 30% NHS while all
other strains were susceptible to killing, especially B. ubonensis MSMB108. This is in agreement with prior studies showing serum sensitivity of B. pseudomallei strains expressing type B2 or rough type O-antigens. Note: Bt, B. thailandensis; Bt-like, B. thailandensis-like species; Bu, B. ubonensis; Bok, B. oklahomensis; and B.sp, Burkholderia sp Discussion https://www.selleckchem.com/products/H-89-dihydrochloride.html O-antigen type A has been described as a disaccharide glucose-talose repeat in B. pseudomallei, B. mallei, and B. thailandensis and these structures differ only by side group modification. B. pseudomallei modifies the talose residue with a 2-O methyl/4-O acetyl group or with a 2-O acetyl/4-O hydroxyl group [15, 16]. In B. mallei, regardless of whether the 2-O position is methylated or acetylated, the 4-O position remains in its native hydroxyl state [13]. B. thailandensis has been reported to have the same modification patterns as B. pseudomallei[12, 14, 22], but a recent study by Ngugi, et al.,[10] suggests that B. thailandensis E264 features a different pattern. Utilizing gas chromatography/mass spectrometry (GC/MS) to examine methylation patterns, they
concluded this strain does not methylate the 2-O position. Brett, et al.,[14] generated mutants of oacA, the 4-O acetyltransferase gene, which also had the unexpected result of a lack of methylation at the 2-O position. This suggests that CHIR-99021 the methyl group may be lost during GC/MS or the E264 strain utilized by Ngugi, et al.,[10] may have undergone mutation in oacA, losing its methylase capabilities. In our current study, 21 out of 23 B. mallei strains expressed intact type A O-antigens while the remaining two (NU7441 cell line ATCC10399 and NCTC120) were rough. Two previous studies showed that B. mallei ATCC10399 had a full ladder pattern by silver staining and immunoblotting [13, 20]. Our genomic analysis has shown that wbiG gene which is known to be involved in the biosynthesis of the type A O-antigen, was disrupted in this strain by IS407A.