| Project/Area Number |
11470380
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Morphological basic dentistry
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| Research Institution | KYUSHU UNIVERSITY (2000)
Meikai University (1999) |
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Principal Investigator |
HANAZAWA Shigemasa Faculty of Dental Sci. KYUSHU UNIVERSITY Prof., 大学院・歯学研究院, 教授 (60060258)
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| Co-Investigator(Kenkyū-buntansha) |
TAKESHITA Akira Meikai Univ. School. of Dent. Assoc., 歯学部, 助手 (70236454)
SHIOTA Susumu Faculty of Dental Sic. KYUSHU UNIVERSITY Ass. Prof., 大学院・歯学研究院, 助手 (00150467)
AMANO Shigeru Meikai Univ. School. of Dent. Assoc. Prof., 歯学部, 助教授 (90167958)
菊地 寛高 明海大学, 歯学部, 助手 (70234193)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥7,300,000 (Direct Cost: ¥7,300,000)
Fiscal Year 2000: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1999: ¥3,800,000 (Direct Cost: ¥3,800,000)
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| Keywords | adult periodontitis / fimbriae / recombination / macrophages / active domain / gene expression / Cytokine / bone resorption / P.gingivalis / レセプター / 遺伝子組み換え / Fibronectin |
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| Research Abstract |
Porphyromonas gingivalis (P.gingivalis) fimbriae play an important role as virulence factors in periodontal disease. To understand the precious biological activities and their active domains of the fimbriae, it is very useful to prepare soluble recombinant fimbrillin, a major subunit protein of the fimbriae. In this study, we developed a procedure to generate an intracellular soluble recombinant fimbrillin (rFimA) in Esherichia coli (E.coli) transformed with fimA gene-inserted expression vector with enterokinase(EK) recognition site at the N terminus and V5-His tag at the C terminus. The rFimA fusion protein consisting of mature regions of FimA and plasmid-derived peptide was visualized at a 43kDa band on SDS-PAGE, and was recognized by the specific monoclonal antibody against P.gingivalis fimbriae. Interestingly, the soluble rFimA fusion protein stimulated bone resorption in our assay system using mouse embryonic calvarial bone cells, and the stimulatory activity of the soluble rFimA fusion protein was 20 times higher than that of purified natural fimbriae.In addition, the rFimA fusion protein induced gene expression of a key molecule, osteoclast differentiation factor(ODF/ OPGL/ RANKL/ TRANCE), involved in osteoclast differentiation, gene. The present study is first demonstration of preparation of P.gingivalis soluble recombinant fimbrillin that displays high stimulatory activity of bone resorption.
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