Gene cloning of Porphyromonas gingivalis specific antigens recognized by serum of adult periodontitis patient.
1. Porphyromonas gingivalis is believed an important pathogen of adult periodontitis. A gene library of P. gingivalis 381 was constructed in lambda phage vector L47.1. The library was probed with serum obtained from patients of severe adult periodontitis. Two clones, lambda MDBG101 and lambda MDBG103 which were expressed, 200 and 160 kDa respectively, were selected and further studied. 2. The expressed antigens in these two clones were also reacted with rabbit antiserum against whole cells, capsular fraction and cell surface fraction of P. gingivalis. 3. Genes coding protein antigens in lambda MDBG101 and lambda MDBG103 were subcloned into high-copy-number plasmid vector pACYC184 and subclones obtained were designated as MD101 and MD103. Recombinant plasmids, pMD101 and pMD103, differed in their restriction endonuclease digestion. 4. Immunodiffusion analysis showed that cloned proteins from MD101 and MD103 reacted with antiserum against P. gingivalis but did not react with antiserum against Prevotella intermedia, Prevotella loescheii and Prevotella asaccharolyticus. 5. These data suggest that P. gingivalis species-specific antigens has been successfully cloned and expressed in Escherichia coli. Since these cloned specific antigens were recognized by adult periodontitis patient sera, the recombinant antigen will be useful material for the development of serodiagnosis of P. gingivalis infection in adults periodontitis.
Hayakawa
M
M
Department of Biochemistry, Nihon University School of Dentistry, Chiba, Japan.
Abiko
Y
Y
Ito
T
T
Sasahara
H
H
Yamano
H
H
Takiguchi
H
H
D000328:Adult; D000942:Antigens, Bacterial / Q000097:blood / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D005779:Immunodiffusion; D007124:Immunoenzyme Techniques; D010518:Periodontitis / Q000276:immunology*; D010957:Plasmids; D016966:Porphyromonas gingivalis / Q000235:genetics* / Q000276:immunology; D013045:Species Specificity
D000942:Antigens, Bacterial; D004262:DNA Restriction Enzymes