Molecular analysis of the gene encoding a protein component of the Eikenella corrodens adhesin complex that is close to the carbohydrate recognition domain |
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Authors: | Hiroyuki Azakami Hiromichi Yumoto Hideaki Nakae Takashi Matsuo and Shigeyuki Ebisu |
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Institution: | Department of Conservative Dentistry, Tokushima University School of Dentistry, Kuramoto-cho 3-18-15, Tokushima 770, Japan |
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Abstract: | A monoclonal antibody against a lectin-like substance (LS) of Eikenella corrodens (Ec) was used for screening the Ec DNA library. Three positive clones that carried an identical 12-kb segment were obtained. A 25-kDa protein, which specifically binds to the antibody, was overproduced in all of the Escherichia coli clones. Deletion analysis showed that the gene encoding the 25-kDa protein was located within a 1.2-kb segment. The nucleotide (nt) sequence of this segment contained an open reading frame encoding a protein of 24 600 Da. We purified the 25-kDa protein from the cloned E. coli strain. The sequence of the first 10 amino acids (aa) from the N-terminus of the purified 25-kDa protein agreed with that deduced from the nt sequence. Since the monoclonal antibody used in this study inhibits the physiological activity of EcLS, we concluded that the 25-kDa protein is a component of the adhesin complex, which is located near the carbohydrate recognition domain of lectin in EcLS. |
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Keywords: | Periodontal disease Monoclonal antibody Gene cloning DNA sequencing Outer membrane protein |
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