| Project/Area Number |
04660324
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
基礎獣医学
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| Research Institution | THE UNIVERSITY OF TOKUSHIMA |
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Principal Investigator |
YAMATO Masayuki THE UNIVERSITY OF TOKUSHIMA, SCHOOL OF MEDICINE, Univ.of Tokushima RESEARCH ASSOCIATE, 医学部, 助手 (90210492)
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| Co-Investigator(Kenkyū-buntansha) |
HIROTA Katuhiko THE UNIVERSITY OF TOKUSHIMA, SCHOOL OF DENTISTRY, Univ.of Tokushima RESEARCH ASS, 歯学部, 助手 (60199130)
ONO Tuneko THE UNIVERSITY OF TOKUSHIMA, SCHOOL OF DENTISTRY, Univ.of Tokushima LECTURER, 歯学部, 講師 (40035514)
FUKUI Koumei THE UNIVERSITY OF TOKUSHIMA, SCHOOL OF DENTISTRY, Univ.of Tokushima PROFESSOR, 歯学部, 教授 (40035407)
TAKEOKA Aya THE UNIVERSITY OF TOKUSHIMA, SCHOOL OF MEDICINE, Univ.of Tokushima TECHNICIAN, 医学部, 教務員 (00116831)
OTA Fusao THE UNIVERSITY OF TOKUSHIMA, SCHOOL OF MEDICINE, Univ.of Tokushima PROFESSOR, 医学部, 教授 (90035478)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1993: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1992: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Monoclonal antibody / Group E streptococci / Str.porcinus / Surface antigen / GC content / Hybridization / Genotype / EIA |
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| Research Abstract |
Monoclonal antibodies(MAbs II-T, IV21 and V16) to surface antigen of group E streptococci (GES) were prepared by fusing myeloma cells with spleen cells of mice immunized with whole cells of GES.One of them, MAb II-T, reacted in an enzyme immunoassay (EIA) with whole cells of serotypes II and V strains, and in a gel diffusion test with autoclaved-saline extracts of both serotypes. MAb IV21 reacted specifically with a serotype IV strain. On the other hand, MAb V16 reacted with almost all strains of GES and serotype e strains of S.mutans. The crude antigen was extracted from whole cell of GES by autoclaving. The extract was purified by DEAE-Sephadex A-25, followed by the treatment with Proteinase K, and further by chromatography with a Sephadex G-200 column. The purified antigen for MAb II-T was a polysaccharide (PS) composed of 98.6% carbohydrate and 1.4% protein with no detectable phosphorus. That for MAb V16 was also found to consist of 92.4% carbohydrate and 7.6% protein. The antigen for MAb IV21 is now being purified. Furthermore, DNA was extracted from 6 serotypes strains (II, IV, V, VI, VII, and VIII) and 3 clinical isolates of GES.The GC contents(%) of these DNAs were determined to be 36 to 38%, indicating that there is no clear difference in the values among 6 serotypes. In DNA-DNA hybridization experiments using radiolabelled DNA of serotype II strain as a probe, the DNA of only serotype V strain showed 100% homology to the probe while others were 56 to 76% homologous. The ribo-typing of GES in now in progress.
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