Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Reexamination Certificate
1999-05-27
2001-01-09
Bui, Phuong T. (Department: 1645)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
C435S007320, C435S069100, C435S243000, C530S350000, C530S820000, C530S825000
Reexamination Certificate
active
06171805
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to a cell surface polypeptide of
Porphyromonas
(Bacteroides)
gingivalis
(another name; periodontopathic organism: hereinafter, referred to as
Porphyromonas gingivalis
) having a molecular weight of about 72-kDa (kilodalton) (hereinafter abbreviated as periodontopathic organism cell surface polypeptide), a DNA encoding the polypeptide, a recombinant DNA wherein the DNA is integrated and a microorganism containing the recombinant DNA.
The DNA obtained in the invention can be utilized as a reagent, namely as a probe for detecting
Porphyromonas gingivalis
. Further, the periodontopathic organism cell surface polypeptide obtained in the invention and produced by a microorganism can be utilized for a diagnosis of patients suffering with periodontal diseases and having antibodies against
Porphyromonas gingivalis
, as an antigen for preparing an antibody for diagnosing patients suffering with periodontal diseases and having
Porphyromonas gingivalis
or as a prophylactic vaccine for periodontal diseases.
2. Description of Related Art
It is reported by Yoshimura et al. in Infect. Immun., 57, 3646 (1989) that since the periodontopathic organism cell surface polypeptide exhibits strong immune response to the sera of patients of periodontal diseases, infecting with
Porphyromonas gingivalis
, antibodies against the polypeptide existing in the sera of the patients can be detected using it. Further, it is reported by Ogawa et al. in Nippon Saikin-gaku Zasshi, 44, 329 (1989), by Yoshimura et al. in J. Bacteriol., 160, 949 (1984) and by Yamaji et al. in Nisshishushi, 33, 349 (1991) that the immune response of the polypeptide is different from that of the known fimbrial protein from the known periodontopathic organism having a molecular weight of 41 kDa, and thus the periodontopathic organism cell surface polypeptide of the invention is different from the ciliary protein.
As to the structure of the periodontopathic organism cell surface polypeptide, its amino acid composition is reported by Yoshimura et al. in Infect. Immun. 57, 3646 (1989), and the amino-terminal amino acid sequence of a purified polypeptide thereof is reported by Yoshimura et al. in FEMS Microbiol. Lett., 92, 47 (1992). Elucidated amino acids are 40 amino acids, and when we compare 40 amino acid with an amino acid number estimated from the molecular weight of the periodontopathic organism cell surface polypeptide, the rate is only under 10%. Further, concerning with a DNA encoding the periodontopathic organism cell surface polypeptide, it is reported by Yoshimura et al. in FEMS Microbiol. Lett., 92, 47 (1992) that its part was obtained. However the base sequence of the gene is not elucidated at all.
The periodontopathic organism cell surface polypeptide is recovered from
Porphyromonas gingivalis
cells, but its supply amount is limited because of difficulty of the culture of
Porphyromonas gingivalis
. Thus development of an industrial process for supplying the periodontopathic organism cell surface polypeptide is desired.
SUMMARY OF THE INVENTION
According to the invention, there are provided a cell surface polypeptide of
Porphyromonas gingivalis
having an amino acid sequence shown in SEQ ID NO: 1 (Numbers 1-558), a DNA encoding the cell surface polypeptide of
Porphyromonas gingivalis
having the amino acid sequence shown in SEQ ID NO: 1 (Numbers 1-558), a recombinant DNA which is a DNA having integrated thereinto the DNA encoding the cell surface polypeptide of
Porphyromonas gingivalis
having the amino acid sequence shown in SEQ ID NO: 1 (Numbers 1-558), and a microorganism having the recombinant DNA which is the DNA having integrated thereinto the DNA encoding the cell surface polypeptide of
Porphyromonas gingivalis
having the amino acid sequence shown in SEQ ID No. 1 (Numbers 1-558).
Further, the DNAs encoding the cell surface polypeptide of
Porphyromonas gingivalis
having the amino acid sequence shown in SEQ ID NO: 1 (Numbers 1-558) according to the present invention include those wherein one or plural bases are added, deleted or replaced by site-directed mutagenesis (Nucleic Acid Research, 10, 6487-6508 (1982)). Therefore, a recombinant DNA wherein a DNA obtained by the mutagenesis is integrated, and a microorganism having the recombinant DNA are also included in the invention. Since a polypeptide produced by recombinant technique using a DNA wherein in the DNA encoding the cell surface polypeptide of
Porphyromonas gingivalis
having the amino acid sequence shown in SEQ ID NO: 1 (Numbers 1-558), one or plural bases are added, deleted or replaced, is a polypeptide wherein in the cell surface polypeptide of
Porphyromonas gingivalis
having the amino acid sequence shown in SEQ ID NO: 1 (Numbers 1-558), one or plural amino acids are added, deleted or replaced, this polypeptide is also included in the invention. A DNA sequence encoding the amino acid sequence shown in SEQ ID No: 1 is shown in SEQ ID No: 3, wherein bases 76-1749 encode amino acids 1-558.
REFERENCES:
Rudinger et al “Peptide Hormone”, edited by Parson, University Park Press, Jun. 1976.
Burgess et al. J Cell Biol, 111:2129-2138, 1990.
De Nardin et al Infect Immun 59(12), 4363-70 (abstract only), 1991.
T. Ogawa et al., “Molecular Cloning and Characterization of the Genes Encoding the Immunoreactive Major-Cell Surface Proteins ofPorphyromonas gingivalis”, FEMS Microbiology Letters, vol. 120, No. 1-2, pp. 23-30, 1994.
K. Watanabe et al., “Molecular Cloning and Expression of a Major Surface Protein (the 75-kDa protein) ofPorphyromonas (Bacteroides) gingivalisinEscherichia coli”, FEMS Microbiology Letters, vol. 92, No. 1, pp. 47-56, 1992.
R. T. Evans et al., Immunization withPorphyromonas (Bacteroides) gingivalisFimbriae Protects against Peridontal Destruction, Infection and Immunity, vol. 60, No. 7, pp. 2926-2935, Jul., 1992.
Fukui Masanori
Hasegawa Mamoru
Hokukoku Shusaburo
Mori Hideharu
Ogawa Tomohiko
Bui Phuong T.
Kyowa Hakko Kogyo Co. Ltd.
Lee Li
Wenderoth , Lind & Ponack, L.L.P.
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