| Project/Area Number |
13470449
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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 |
矯正・小児・社会系歯学
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| Research Institution | Nagasaki University (2002)
Osaka University (2001) |
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Principal Investigator |
FUJIWARA Taku Nagasaki University, Graduate School of Biomedical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (00228975)
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| Co-Investigator(Kenkyū-buntansha) |
OOSHIMA Takashi Osaka University, Graduate School of Dentistry, Professor, 大学院・歯学研究科, 教授 (80116003)
FUKUMOTO Satoshi Nagasaki University, Graduate School of Biomedical Sciences, Instructor, 大学院・医歯薬学総合研究科, 助手 (30264253)
KUBOTA Kazumi Nagasaki University, Dental hospital, Assistant Professor, 歯学部附属病院, 講師 (30240914)
SHINNTANI Seikou Osaka University, Graduate School of Dentistry, Associate Professor, 大学院・歯学研究科, 助教授 (90273698)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥16,600,000 (Direct Cost: ¥16,600,000)
Fiscal Year 2002: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 2001: ¥12,100,000 (Direct Cost: ¥12,100,000)
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| Keywords | DNA vaccine / Streptococcus mutans / Glucosyltransferase / Dental caries / S.mutans / 哺乳類発現ベクター |
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| Research Abstract |
Streptococcus mutans possesses three glucosyltransferase (GTF) enzymes that coordinate to synthesize adhesive glucan, which facilitates adhesion and accumulation of S. mutans cells to tooth surfaces. Thus, GTFs are considered to be a major virulence factor of dental caries. We constructed DNA vaccines against GTFs and then attempted to determine their immunogenicity. Two functional domains of GTFs, the catalytic (CAT) and glucan-binding (GBD) domains, were used as targets of the DNA vaccines.
DNA fragments coding these domains were amplified by PCR and ligated into an eukaryotic expression vector, pcDNA3, which resulted in the DNA vaccine plasmids pcDNA3/CAT and pcDNA3/GBD. A mouse embryo cell line, NIH 3T3, was transfected with either pcDNA3/CAT or pcDNA3/GBD and the expressions of mRNA and recombinant proteins for CAT or GBD in the transfected cells were subjected to reverse transcription-PCR (RT-PCR) and Western blot analyses, respectively. Using RT-PCR, CAT-and GBD- specific mRNA was detected in pcDNA3/CAT and pcDNA3/GBD transfected cells, while Western blot analysis also revealed that the CAT- and GBD- specific recombinants were expressed in the respective cells. Our results indicated that these DNA vaccines expressed recombinant proteins as antigens in mammalian cells and possessed the ability to induce immune responses.
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