| Project/Area Number |
14570387
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Legal medicine
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| Research Institution | National University Corporation Tottori University |
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Principal Investigator |
IRIZAWA Yoshito National University Corporation Tottori University, Faculty of Medicine, Professor, 医学部, 教授 (90112226)
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| Co-Investigator(Kenkyū-buntansha) |
YUASA Isao National University Corporation Tottori University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (00093633)
NAKAGAWA Mayumi National University Corporation Tottori University, Faculty of Medicine, Associate Research, 医学部, 助手 (00243410)
UMETSU Kazuo National University Corporation Yamagata University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (10091828)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2003: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | orosomucoid / alpha-1-acid glycoprotein / gene diversity / gene duplication / function analysis / α1-酸性糖タンパク / α_1-酸性糖タンパク |
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| Research Abstract |
In this study, we have clarified the genomic structure of the ORM gene in African and European populations. They showed duplication of the ORM1^*F1 gene, i.e., ORM1^*F1-ORM1^*F1-ORM2^*M structure. This major triplicated gene was evidently different from the genomic structure observed in Japanese populations, which have ORM1^*F1-ORM1^*S1-ORM2^*M structure. In Africans ORM1A^*S1(1)-ORM1B^*S2(T)-ORM2^*M was also observed, while in Europeans ORM1^*F2-ORM1^*S1-ORM2^*M structure was also found. Although independent recombination events have occurred in Africans and Japanese, recombination breakpoints lay within a small genomic interval around exon 1 of the ORM1B gene in both populations. A duplication of the ORM2 gene was rather rare. The highest nucleotide diversity of the ORM2 gene was found in Africans.
Subsequently, we have cloned a cDNA of an ORM1 gene and introduced mutations in the cloned cDNA by site-directed mutagenesis. And then the mutated genes were ligated into commercial available pcDNA3.1 expression vector. After subcloning in E.coli and transfecting into Cos-7 cell, we have obtained mutated proteins. Thus, we established the method for the preparation of mutated ORM1 proteins. This method and these proteins would be useful for studying the role of the ORM protein in transportation of basic drugs.
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