| Project/Area Number |
03557028
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Hygiene
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| Research Institution | HOKKAIDO UNIVERSITY |
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Principal Investigator |
KOJIMA Yutaka Hokkaido Univ., Grad. School of Env. Earth Sci., Prof., 大学院・地球環境科学研究科, 教授 (50135555)
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| Co-Investigator(Kenkyū-buntansha) |
NISHINO Kohsuke Doshisha Women's College of Liberal Arts, Faculty of Human Life and Sci., Prof., 生活科学部, 教授 (20093289)
KURASAKI Masaaki Hokkaido Univ., Grad. School of Env. Earth Sci., Inst., 大学院・地球環境科学研究科, 助手 (80161727)
NIIOKA Tadashi Hokkaido Univ., Grad. School of Env. Earth Sci., Assoc. Prof., 大学院・地球環境科学研究科, 助教授 (20123953)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1993: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1991: ¥4,100,000 (Direct Cost: ¥4,100,000)
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| Keywords | metallothionein / metal-binding protein / stress / ELISA / monoclonal antibody / evaluation of stress / human metallothionein / Radioimmunoassay / メタロチオネイン遺伝子 / メタロチオネイン抗体 / モノクロナル抗体 / 大腸菌での発現 / ヒト・メタロチオネインII |
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| Research Abstract |
It has known that the synthesis of metallothioneins (MTs), metal-binding proteins, is induced by various physical and psychological stressors resulting in increases in MT contents in the liver, blood, and urine. The goal of this study is to establish quantitative evaluation of stress using metallothionein induced under stressful conditions, that is, it is to establish objective evaluation of human stress with high accuracy using MT as an indicator of stress. To establish the evaluation, firstly monoclonal anti MT antibody is to be obtained using human MT, which is used as the antigen, obtained abundantly by a genetic-engineering method. Secondly, MT contents in urine or blood is to be measured by an ELISA method using the anti MT antibody. The results obtained are as follows :
1) we successfully obtained the results of MT gene expression by introduction of the human MT II gene into Escherichia coli.
2) we obtained abundant refined MT by gel filtration chromatography and high-performance-liquid-chromatography on the crude MT obtained from mass culture of the Escherichia coli.
3) we obtained monoclonal anti-MT antibody by cell fusion of mouse myeloma cell and spleen cell of a mouse injected with the human MT II.
4) an ELISA method using the monoclonal anti-MT antibody is being developed.
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