title: Immunologic and genetic characterization of lipooligosaccharide variants in a Neisseria meningitidis serogroup C strain.
abstract: Neisseria meningitidis shows great variation in expression of structurally different lipooligosaccharides (LOS) on its cell surface. To better understand the LOS diversity that may occur within an individual strain, a group C wild-type strain, BB305-Tr4, and two stable isogenic LOS variants, Tr5 and Tr7, were selected for this study. SDS-PAGE analysis showed a size reduction of Tr5 and Tr7 LOS compared to that of Tr4. Immunoblotting showed that parental Tr4 LOS reacted with L1, L2 and L3,7 antibodies, variant Tr5 LOS with L1 and L6 antibodies, while Tr7 LOS was non-typeable. Genetic analysis showed that the gene organization at the lgt-1 locus in the three strains was lgtZ,C,A,B,H4 in Tr4, lgtZ,C,A,H4 in Tr5 and lgtZ,C,A,H9 in Tr7. The genetic differences in the three strains were consistent with their phenotypic changes. Sequence comparison revealed two independent recombination events. The first was the recombination of repeated DNA fragments in the flanking regions to delete lgtB in Tr5. The second was the recombination of a fragment of two genes, lgtB and lgtH4, to create an inactive lgtH9 allele with a mosaic structure in Tr7. These findings suggest that besides phase variation, homologous recombination can contribute to the genetic diversity of the lgt locus and to the generation of LOS variation in N. meningitidis.
authors.affiliation: Laboratory of Bacterial Polysaccharides, Center for Biologics Evaluation and Research, FDA, 8800 Rockville Pike, Bethesda, MD 20892, USA. zhu@cber.fda.gov
authors.forename: Peixuan
authors.initials: P
authors.lastname: Zhu
authors.forename: Chao-Ming
authors.initials: CM
authors.lastname: Tsai
authors.forename: Carl E
authors.initials: CE
authors.lastname: Frasch
mesh_terms: D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D014644:Genetic Variation; D016695:Glycosyltransferases / Q000235:genetics*; D008070:Lipopolysaccharides / Q000276:immunology*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D038542:Neisseria meningitidis, Serogroup C / Q000235:genetics* / Q000276:immunology* / Q000472:pathogenicity; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D012689:Sequence Homology, Nucleic Acid; D014166:Transferases / Q000032:analysis
chemical_list: D008070:Lipopolysaccharides; C023023:lipid-linked oligosaccharides; D014166:Transferases; D016695:Glycosyltransferases; C042308:phosphatidylglycerol glyceryl transferase