Gene banks of chromosomal DNA from Bacteroides gingival is 381 were constructed utilizing the bacteriophage replacement vector λCharon4A. A clone encoding a protein antigen from B. gingivalis was identified by Western-blot screening, with use of antiserum induced to extracts of B. gingivalis cells. DNA fragments from the phage clone were subcloned into the plasmid vector pACYC184 to yield an immunoreactive clone. Cell extracts from the subclone reacted with antiserum against B. gingivalis, but did not react with antisera to B. asaccharolyticus, B. intermedius, or B. melaninogenicus. The antiserum against the purified clone products reacted with N-lauryl sarcosine extracts from B. gingivalis cells, but did not react with those of other Bacteroides cells. In addition, human serum from periodontitis patients reacted with the clone product by Western electrophoretic transfer and immunoblotting analysis. These data suggest that the gene coding for a B. gingivalis-specific protein antigen was successfully cloned and functionally expressed in Escherichia coli. This clone product may prove useful for further studies of B. gingival is as a periodontal pathogen.
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References
1.
Abiko, Y. and Takiguchi, H. (1986): Cloning and Expression of Bacteroides gingivalis Antigen Gene into Escherichia coli, J Dent Res65:737, Abstr. No. 106 (IADR).
2.
Dickinson, D.P.; Kubiniec, M.; Yoshimura, F.; and Genco, R.J. (1986): Molecular Cloning of B. gingivalis Fimbrial Protein Subunit Gene, J Dent Res65:737, Abstr. No. 105 (IADR).
3.
Doty, S.L.; Lopatin, D.E.; Syed, S.A.; and Smith, F.N. (1982): Humoral Immune Response to Oral Microorganisms in Periodontitis, Infect Immun37:499-505.
4.
Ebersole, J.L.; Taubman, M.A.; Smith, D.J.; and Frey, D. (1986): Human Immune Responses to Oral Microorganisms: Patterns of Systemic Antibody Levels to Bacteroides Species, Infect Immun51:507-513.
5.
Hanazawa, S.; Saitoh, K.; Ohmori, Y.; Nishihara, H.; Fuji-Wara, S.; and Kitano, S. (1984): Production of Monoclonal Antibodies that Recognize Specific and Cross-reactive Antigens of Bacteroides gingivalis, Infect Immun46:285-287.
6.
Hohn, B. and Hohn, T. (1974): Activity of Empty, Headlike Particles for Packaging of DNA of Bacteriophage in vitro, Proc Natl Acad Sci USA71 :2372-2376.
7.
Kikuchi, Y.; Kiyoshige, T.; Sasaki, S.; Okuda, K.; Kato, T.; Naito, Y.; Ono, M.; and Takazoe, I. (1985): Protective Efficacy of Antiserum Against Bacteroides gingivalis, J Jpn Assoc Periodont27:106-113.
8.
Mansheim, B.J.; Stenstrom, M.L.; Low, S.B.; and Clark, W.B. (1980): Measurement of Serum and Salivary Antibodies to the Oral Pathogen Bacteroides asaccharolyticus in Human Subjects, Arch Oral Biol25:553-557.
9.
Mouton, C.; Hammond, P.G.; Slots, J.; and Genco, R.J. (1981a): Serum Antibodies to Oral Bacteroides asaccharolyticus (Bacteroides gingivalis) : Relationship to Age and Periodontal Disease, Infect Immun31 :182-192.
10.
Mouton, C.; Hammond, P.G.; Slots, J.; Reed, M.J.; and Genco, R.J. (1981b): Identification of Bacteroides gingivalis by Fluorescent Antibody Staining, Ann Microbiol (Inst. Pasteur)132B:69.
11.
Naito, Y.; Okuda, K.; and Takazoe, 1. ( 1984): Immunoglobulin G Response to Subgingival Gram-negative Bacteria in Human Subjects, Infect Immun45:47-51.
12.
Naito, Y.; Okuda, K.; and Takazoe, 1. ( 1985): Monoclonal Antibodies against Surface Antigens of Bacteroides gingivalis, Infect Immun50:231-235.
13.
Naito, Y.; Okuda, K.; and Takazoe, I. (1987): Detection of Specific Antibody in Adult Periodontitis Sera to Surface Antigens of Bacteroides gingivalis, Infect Immun55:832-834.
14.
Okuda, K.; Naito, Y.; Ohta, K.; Fukumoto, Y.; Kimura, Y.; Ish-Ikawa, Y.; Kinoshita, S.; and Takazoe, 1. ( 1984): Bacteriological Study of Periodontal Lesions in Two Sisters with Juvenile Periodontitis and Their Mother, Infect Immun45:118-121.
15.
Okuda, K.; Kato, T.; Naito, Y.; Ono, M.; Kikuchi, Y.; and Takazoe, I. (1986): Susceptibility of Bacteroides gingivalis to Bactericidal Activity of Human Serum, J Dent Res65:1024-1027.
16.
Reed, M.J.; Slots, J.; Mouton, C.; and Genco, R.J. (1980): Antigenic Studies of Oral and Non-oral Black-pigmented Bacteroides Strains, Infect Immun29:564-574.
17.
Schei, 0.; Waerhaug, J.; Lovdal, A.; and Arso, A. (1959): Alveolar Bone Loss as Related to Oral Hygiene and Age, J Periodontol30:7-16.
18.
Schenck, K. (1985): IgG, IgA and IgM Serum Antibodies Against Lipopolysaccharide from Bacteroides gingivalis in Periodontal Health and Disease, J Periodont Res20:368-377.
19.
Schenkein, H.A. and Genco, R.J. (1977a): Gingival Fluid and Serum in Periodontal Diseases. I. Quantitative Study of Immunoglobulins, Complement and Other Plasma Proteins, J Periodontol48:772-777.
20.
Schenkein, H.A. and Genco, R.J. (1977b): Gingival Fluid and Serum in Periodontat Diseases. II. Evidence for Cleavage of Complement Components C3, C3 Pro-activator (Factor B) and C4 in Gingival Fluid, J Periodontol48:778-784.
21.
Skalka, A. and Shapiro, L. (1976): In situ Immunoassays for Gene Translation Products in Phage Plaques and Bacterial Colonies, Gene1 :65-79.
22.
Slots, J. (1979): Subgingival Microflora and Periodontal Disease , J Clin Periodontol6:351-382.
23.
Slots, J. (1982): Importance of Black-pigmented Bacteroides in Human Periodontal Disease. In: Host-Parasite Interactions in Periodontal Disease, R.J. Genco and S.E. Mergenhagen, Eds., Washington, D.C. : American Society for Microbiology, pp. 27-45.
24.
Smith, D.J.; Gadalla, L.M.; Ebersole, J.L.; and Taubman, M.A. (1985): Gingival Crevicular Fluid Antibody to Oral Microorganisms. III. Association of Gingival Homogenate and Gingival Crevicular Fluid Antibody Level, J Periodont Res20:357-367.
25.
Socransky, S.S. (1977): Microbiology of Periodontal Disease-Present Status and Future Consideration, J Periodontol48:497-504.
26.
Socransky, S.S.; Tanner, A.C.R.; Haffajee, A.D.; Hillman, J.D.; and Goodson, J.M. (1982): Present Status of Studies on the Microbial Etiology of Periodontal Diseases. In: Host-Parasite Interactions in Periodontal Diseases, R.J. Genco and S.E. Mergenhagen, Eds., Washington, D.C.: American Society for Microbiology , pp. 1-12.
27.
Spiegel, C.A.; Hayduk, S.E.; Minah, G.E.; and Krywolap, G.N. (1979): Black-pigmented Bacteroides from Clinically Characterized Periodontal Sites, J Periodont Res14:376-382.
28.
Taubman, M.A.; Ebersole, J.L.; and Smith, D.J. (1982): Association between Systemic and Local Antibody and Periodontal Disease. In: Host-Parasite Interactions in Periodontal Disease, R.J. Genco and S.E. Mergenhagen, Eds., Washington, D.C.: American Society for Microbiology , pp. 283-298.
29.
Tew, J.G.; Marshall, D.R.; Moore, W.E.C.; Best, A.M.; Palcanis, K.G.; and Ranney, R.R. (1985): Serum Antibody Reactive with Predominant Organisms in the Subgingival Flora of Young Adults with Generalized Severe Periodontitis , Infect Immun48:303-311.
30.
Tolo, K. and Brandtzaeg, P. (1982): Relation Between Periodontal Disease Activity and Serum Antibody Titers to Oral Bacteria. In: Host-Parasite Interactions in Periodontal Disease, R.J. Genco and S.E. Mergenhagen, Eds., Washington, D.C.: American Society for Microbiology, pp. 270-282.
31.
White, D. and Mayrand, D. (1981): Association of Oral Bacteroides with Gingivitis and Adult Periodontitis, J Periodont Res16:259-265.
