| Project/Area Number |
62480155
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
細菌学
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| Research Institution | The University of Tokushima |
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Principal Investigator |
KAWATA Tomio The University of Tokushima, School of Medicine, 医学部, 教授 (60035368)
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| Co-Investigator(Kenkyū-buntansha) |
TAKEOKA Aya The University of Tokushima, School of Medicine, 医学部, 教務員 (00116831)
YAMATO Masayuki The University of Tokushima, School of Medicine, 医学部, 助手 (90210492)
KOGA Tetsuro The University of Tokushima, School of Medicine, 医学部, 助手 (20093859)
TAKUMI Kenji The University of Tokushima, School of Medicine, 医学部, 助教授 (90035428)
増田 邦義 徳島大学, 医学部, 助手 (60035483)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1988: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1987: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | Bacterial surface / Cell wall / Protein layer / Molecular organization / Immunochemical characterization / Biological activity / Clostridium difficile / Clostridium botulinum / ラクトバシルス・アシドフィルス |
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| Research Abstract |
We have investigated the molecular organization, immunochemical properties, and biological activities of the regularly arranged proteins (RA proteins) in the outer cell well layers of some clostridia and lactobacilli. 1) The cell wall of clostridium difficile was revealed by electron microscopy to have an outer layer composed of a nearly square array and contained the two major proteins with Mrs of 32 and 45 kDa, or 38 and 42 kDa, depending on the strains. The amino acid composition and peptide maps of the two RA proteins differed from each other. Ca^<2+> was essential for the reassembly of regular array from the two RA proteins solubilized with urea or guanidine hydrochloride. The 32 kDa protein possessed the adhesive activity ot HeLa cells, whereas the 45 kDa protein did not. Immunoelectron microscopy revealed that the two RA proteins existed on the cell well surface. 2) A linearly arranged thin array was observed on the outer cell wall layer of Clostridium botulinum type E and composed of three major proteins with Mrs of 90, 60 and 25 kDa. The 90 and 60 kDa proteins were purified and immunochemically characterized. Both proteins were shown to locate on the wall surface by immunoelectron microscopy. 3) Regular arrays were present in only strains belonging to a group of lactobacillus acidophilus but not in the other groups. Based on the peptide mapping and immunoblotting of the RA proteins, the A group could be further classified into 4 or more subgroups.
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