| Project/Area Number |
09558095
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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 | 展開研究 |
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| Research Field |
Molecular biology
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| Research Institution | Nagoya University |
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Principal Investigator |
KIKUCHI Akihiko Nagoya Univ., Sch.Med., Professor, 医学部, 教授 (40283428)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Masato Mitsubishi-Kagaku, Inst.Life.Sci., S.Res., 主任研究員
AKASHI Tomohiro Nagoya Univ., Sch.Med., Res.Associate, 医学部, 助手 (00222513)
田中 眞人 三菱化学, 生命科学研究所, 主任研究員
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| Project Period (FY) |
1997 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥5,200,000 (Direct Cost: ¥5,200,000)
Fiscal Year 2000: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1999: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1998: ¥2,900,000 (Direct Cost: ¥2,900,000)
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| Keywords | DNA topoisomerase / Monoclonal Antibody / Epitopes / Cell Division Cycle / Signal sequence / Nuclear localization / Diagnosis and therapy / Mycology / 機能領域 / たんぱく分解 / 細胞内局在化 / 酵母 / DNAポイソメラーゼ / 活性阻害 / 核移行シグナル / 種特異性 / クリーバブル・コンプレックス / 大腸菌による発現 / トポIIα・トポIIβ / アスペルギルス / 難治性の深在性真菌症 / GFP融合たんぱく |
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| Research Abstract |
(1) Epitope analysis of anti-DNA topoisomerase II monoclonal antibody
We established over 100 hybridoma cells secreting anti-DNA topoisomerase II (topo II). We selected some and determined each epitope of oligopeptide level. One of them (6H8) can recognize topo II of every eucaryotes and another (8D2) is specific to proliferating cells and useful for diagnosis for neoplastic cells.
(2) Analysis of topoisomerase of filamentous fungi and yeast.
We cloned tope II gene of Aspergillus, Penicillium and Candida and analyzed nuclear localization signals at C-terminus region by Ala scanning method. From their sequence data we developed rapid PCR analysis for diagnosis test.
(3) Novel site of localization of human topo II in nuclear region
In addition to nuclear localization signal of human topo IIβ, we detected a novel signal sequence for the distinct localization site close to nucleolus. This signal is also found in several proteins such as RNA binding proteins and NPI46 (nucleolar peptidyl prolyl cis trans isomerase).
(4) The monoclonal antibodies against human DNA topoisomerase IIα
extensively used and characterized throughout this research project was filed for patent application by Mitsubishi-Kagaku and now comercialized for the diagnostic reagent of malignant tumor cells.
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