Inconsistency between the fimbrilin gene and the antigenicity of lipopolysaccharides in selected strains of Porphyromonas gingivalis.
Immunochemical specificity of lipopolysaccharide and the molecular property of the gene encoding the fimbrilin (fimA) of Porphyromonas gingivalis strains were examined using 'fimbriated' strains 381 and HG564 and 'non-fimbriated' strains 381FL and W50. Lipopolysaccharide from strains 381, 381FL and HG564 reacted with monoclonal antibody raised to lipopolysaccharide from strain 381 to give a fused precipitin band by the immunodiffusion test. However, silver staining and Western blotting of lipopolysaccharide clearly revealed a difference in profile of bands between strains 381 and 381FL. On the other hand, lipopolysaccharide from W50 formed another precipitin band and reacted with the antibody, but only at higher concentrations of lipopolysaccharide. The fimA genes in these strains were amplified by polymerase chain reaction and cloned. Sequencing of the fimA gene revealed that the fimA(W50) was almost identical to fimA(HG564), but a notable difference was observed at the start codon of the open reading frame, while the fimA(381FL) was considerably different from fimA of other strains and its open reading frame was found to be missing. These results indicate that the molecular structure of the fimA genes of these strains is not homologous, indicating that molecular modifications in the fimA gene should occur during in vitro passages and maintenance of strains of P. gingivalis in laboratories.
Department of Oral Microbiology, Osaka University Faculty of Dentistry, Japan.
T
T
Fujiwara
I
I
Nakagawa
S
S
Morishima
I
I
Takahashi
S
S
Hamada
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003429:Cross Reactions; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000737:chemistry* / Q000276:immunology; D008070:Lipopolysaccharides / Q000737:chemistry / Q000276:immunology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D016966:Porphyromonas gingivalis / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D016415:Sequence Alignment
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D008070:Lipopolysaccharides; C063055:fimbrillin; D036781:Fimbriae Proteins