| Project/Area Number |
01570333
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Legal medicine
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| Research Institution | Yamanashi Medical College |
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Principal Investigator |
OYA Masakazu Yamanashi Medical College, School of Medicine, Professor, 医学部, 教授 (70079979)
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| Co-Investigator(Kenkyū-buntansha) |
KIMURA Yukiko Yamanashi Medical College, School of Medicine, Assistant, 医学部, 助手 (40195384)
KOMATSU Nori Yamanashi Medical College, School of Medicine, Assistant, 医学部, 助手 (80153667)
KIDO Akira Yamanashi Medical College, School of Medicine, Assistant Professor, 医学部, 助教授 (60021440)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1990: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1989: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | Serum Protein Groups / Isoelectric Focusing / Electroblotting / Phenotyping in Seminal Stains / 当電点電気泳動法 |
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| Research Abstract |
Phenotypings of transferrin (TF), group specific component (GC), alpha 2HS-glycoprotein (AHS) and orosomucoid (OR) were attempted in semen and seminal stains by means of isoelectric focusing and electroblotting.
Ejaculates were collected from patients at the Fertility Clinic of Yamanashi Prefectural Hospital and healthy male volunteers. Isoelectric focusing was performed on 0.5 mm polyacrylamide gel using pH 5-7 Ampholine for TF types, pH 4-6 for GC types, pH 3.5-6 for PI types, pH 4.2-5.4 for AHS types and pH 4-6.5 for OR types. After electrofocusing, proteins were transferred onto a sheet of nitrocellulose membrane, then allowed to react with specific antiserum as primary antibody and peroxidase-labeled or alkaline phosphatase-labeled anti-IgG as second antibody. The immune complex formed on the membrane were stained with histochemical technique.
OR1 patterns in semen samples were identical with those in serum samples and there is no difference of detection between transfer methods or staining methods. The results for the distribution of OR1 types in 200 semen samples were similar to those for another Japanese population. The determination limits of OR1 types in 20 seminal stains were about 1 Week. The study of the demonstration of OR2 types in seminal stains is in progress.
GC patterns in semen samples were almost similar to those in serum samples and were detected in spite of low GC activity. The distribution of GC types examined in 200 semen samples is same as those in serum samples from Yamanashi population. GC phenotyping from seminal stains made on filter papers were difficult for the low activities.
Despite the various examinations of the concentration of samples and highly sensitive staining methods, the determinations of TF types, PI types and AHS types from semen samples were difficult for trace amounts of these proteins.
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