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Infection and Immunity, February 1999, p. 942-945, Vol. 67, No. 2
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Cloning of the Gene Encoding the Actinobacillus actinomycetemcomitans Serotype b OmpA-Like Outer Membrane Protein

Hitoshi Komatsuzawa,1,* Toshihisa Kawai,2 Mark E. Wilson,3 Martin A. Taubman,2 Motoyuki Sugai,1 and Hidekazu Suginaka1

Department of Microbiology, Hiroshima University School of Dentistry, Hiroshima, 734-8553 Japan1; Department of Immunology, Forsyth Dental Center, Boston, Massachusetts 02115-37992; and Dental Research Center, University of Medicine and Dentistry of New Jersey, New Jersey Dental School, Newark, New Jersey 07103-24003

Received 22 July 1998/Returned for modification 5 October 1998/Accepted 18 November 1998

The gene encoding an outer membrane protein A (OmpA)-like, heat-modifiable Omp of Actinobacillus actinomycetemcomitans ATCC 43718 (strain Y4, serotype b) was cloned by a PCR cloning procedure. DNA sequence analysis revealed that the gene encodes a protein of 346 amino acid residues with a molecular mass of 36.9 kDa. The protein expressed by the cloned gene reacted with a monoclonal antibody to the previously described 29-kDa Omp (Omp29) of strain Y4. This monoclonal antibody reacted specifically with Omp29 of A. actinomycetemcomitans (serotype b), but not with any Omp of Escherichia coli, including OmpA. This protein exhibited characteristic heat modifiability on sodium dodecyl sulfate-polyacrylamide gels, showing an apparent molecular mass of 29 kDa when unheated and a mass of 34 kDa when heated. The N-terminal amino acid sequence of the protein expressed in E. coli perfectly matched those deduced from the purified Omp29 of strain Y4. The deduced amino acid sequence of the gene coding for Omp29 from serotype b matched completely (except for valine at position 321) that of a recently reported omp34 gene described for A. actinomycetemcomitans serotype c (NCTC 9710). Because of the conserved nature of the gene within these serotypes, we designated the gene described herein from serotype b as omp34.

* Corresponding author. Mailing address: Department of Microbiology, Hiroshima University School of Dentistry, Kasumi 1-2-3, Minami-ku, Hiroshima City, Hiroshima 734, Japan. Phone: 81 82 257 5636. Fax: 81 82 257 5639. E-mail: hkomatsu{at}ipc.hiroshima-u.ac.jp.

Infection and Immunity, February 1999, p. 942-945, Vol. 67, No. 2
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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