| Project/Area Number |
61440040
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Legal medicine
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| Research Institution | Department of Legal Medicine, Shinshu University |
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Principal Investigator |
HASEKURA Hayato Dept of Legal Med, Shinshu Univ., Professor, 医学部, 教授 (40049789)
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| Co-Investigator(Kenkyū-buntansha) |
OTA Masao Dept of Legal Med., assistant Professor, 医学部, 助手 (50115333)
FUKUSHIMA Hirofumi Dept of Legal Med., associate Professor, 医学部, 助教授 (70135218)
米村 勇 信州大学, 法医学教室, 助手 (30020762)
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| Project Period (FY) |
1986 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥19,600,000 (Direct Cost: ¥19,600,000)
Fiscal Year 1989: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1988: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1987: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 1986: ¥9,500,000 (Direct Cost: ¥9,500,000)
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| Keywords | Flow Cytometric Analysis / RH blood groups / genotype and phenotype / Human Leukocyte Antigen / ^<125>I-Protein A / HLA-DNA typing / 遺伝子型 / サザンハイブリダイゼ-ション / HLAークラスII抗原 / RFLP / セル・ソーター / Rh式血液型 / De1型D^u型 / RIA / 赤血球型 / ELISA / 血液型 / フローサイトメトリー / 蛍光標識抗体 / 違伝子型 / dosage effect / 法医遺伝学 / 白血球型 |
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| Research Abstract |
1. Rhl(Rho,D),HLA-A2,HLA-A24,HLA-Bw52 and HLA-DR2 antigen activities have been analyzed by the use of indirect FCM. Various RH blood group genotypes including rare variants expressing Del and D^u(high-grade and low-grade)were successfully determined from the intensity of fluorescence detected in FCM using anti-D IgG fractionated in a Protein A Sepharose CL-4B clolumn as the primary antibody. Also, the HLA genotypes could be successfully determined from the different degrees of fluorescence intensity detected by using FITC(fluorescein isothiocyanate) conjugated anti-human IgG goat F(ab)' serum as the secondary antibody.
2. After sensitization by anti-D serum, the ^<125>-protein A bindings were investigated in D, Del, and d red cells. The relative amount of ^<125>-protein A bound to Del cells was less than one per cent of the D cells.
3. Genomic DNAs from 12 normal individual cells expressing DR1,DR2,DRlDR2,DR2DRW8,DRlDRw8,DRw8,DR1DR9 and DR9 antigens were digested with 3 different restriction endonucleases(EcoRI,MspI and TagI) and hybridized to a DQbeta cDNA probe. Distribution analysis of restriction fragment length polymorphism (RFLP) was able to differentiate the zygosity of DR antigens(a DR homozygote or a DR heterozygote involving DR-gene).
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