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Characterization of recombinant and native forms of a cell surface antigen of Porphyromonas (Bacteroides) gingivalis.

Department of Microbiology, University of British Columbia, Vancouver, Canada.

ABSTRACT

The cloning of genes encoding putative cell surface antigens of Porphyromonas gingivalis ATCC 33277 has been reported previously (B. C. McBride, A. Joe, and U. Singh, Arch. Oral Biol. 55:59S-68S, 1990). This study characterizes the recombinant protein rPgAg1, which is highly expressed in clone BA3, and the corresponding 51-kDa native antigen PgAg1. Cellular localization studies with monospecific antibodies to rPgAg1 in a Western immunoblot assay of a P. gingivalis membrane fraction and immunogold labeling of intact P. gingivalis cells confirmed the cell surface location of the native PgAg1 molecule. The pgag1 gene was found to be present in all four strains of P. gingivalis examined, and the gene product was expressed. Highly homologous DNA sequences and immunologically related proteins, however, were not detected in related species in the group formerly known as black-pigmented Bacteroides. This suggests that PgAg1 is specific to P. gingivalis and is highly conserved within this species. A protein data base search with the NH2-terminal amino acid sequence of rPgAg1 did not identify any significantly similar protein sequences. The high level of expression of rPgAg1 was not dependent on the insertional orientation of the cloned fragment. It therefore appears that a P. gingivalis promoter is present which is well recognized by the transcriptional apparatus of the Escherichia coli cloning host. The promoter element and structural gene for a specific cell surface antigen of P. gingivalis have been cloned.

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