| Project/Area Number |
09044206
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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 |
Bioproduction chemistry/Bioorganic chemistry
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| Research Institution | THE UNIVERSITY OF TOKYO |
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Principal Investigator |
YAMAGUCHI Isomaro DEPARTMENT OF APPLIED BIOLOGICAL CHEMISTRY, THE UNIVERSITY OF TOKYO, PROFESSOR, 大学院・農学生命科学研究科, 教授 (00012013)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAJIMA Masatohi DEPARTMENT OF APPLIED BIOLOGICAL CHEMISTRY, THE UNIVERSITY OF TOKYO, ASSISTANT PROFESSOR, 大学院・農学生命科学研究科, 助手 (50237278)
SUZUKI Yoshihito DEPARTMENT OF APPLIED BIOLOGICAL CHEMISTRY, THE UNIVERSITY OF TOKYO, ASSOCIATE PROFESSOR, 大学院・農学生命科学研究科, 助教授 (90222067)
CONRAD Udo ガータースレーベン植物育種研究所, 主任研究員
WEILER W.Elm ルール大学, 植物生理学科, 教授
WEILER W Elm ルール大学, 植物生理学科, 教授
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥5,300,000 (Direct Cost: ¥5,300,000)
Fiscal Year 1999: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1997: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | gibberellin / sigle chain Fv / anti-idiotypic antibody / dwarf / 植物ホルモン / 形質転換タバコ |
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| Research Abstract |
A monoclonal anti-idiotypic antibody was raised against anti-gibberellin A4 (GA4) antibody, which recoguniaes biologically active GAS Such as GA1 and GA4 specifically. By using the property of the antibody to compete with GA1 and GA4 in binding to anti-GA4-antibody, we successfully applied the anti-idiotypic antibody to ELISA as a tracer for measuring GA1 and GA4. The single chain Fv (scFv) gene of the anti-idiotypic antibody was constructed, and SCFV expressed in E. coli showed binding activity to anti-GA4 antibody.
An anti-GA24/19 scFv gene was constracted from γ and κ genes cloned from a hybridoma cell line producing monoclonal antibody against GA24/19, biosynthetic precursors of GA4/1 which are biologically active per se. The scFv gene was introduced into tobacco plants after the binding activity of the scFv expressed in E. coli was confirmed. When the SCFV expression is targeted to endoplasmic reticulum, the plants could accumulate the scFv protein with the antigen binding activity up to 3.6% of the total soluble protein. On the other hand, when the expression is targeted to cytosol, accumulation of the scFv protein was not detected at all. The dwarf phenotype of the transgenic plants expressing the scFv protein, together with the preliminary analytical data indicating a decreased level of gibberellin A1 in the dwarf transgenics, suggested that the scFv decreased the concentration of bioactive GA by trapping and inhibiting the metabolism of GA24 and/or GA19 to GA4 and/or GA1.
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