| Project/Area Number |
61480444
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory medicine
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| Research Institution | Osaka Medical College |
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Principal Investigator |
INAI Shinya Osaka Medical College Professor, 医学部, 教授 (90131317)
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| Co-Investigator(Kenkyū-buntansha) |
MIKI Tetsuroh Osaka University Medisal School Assistant Professor, 医学部, 助手 (00174003)
MORIYAMA Takeshi Osaka Medical College Assistant Professor, 医学部, 助手 (90182279)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥6,400,000 (Direct Cost: ¥6,400,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥5,400,000 (Direct Cost: ¥5,400,000)
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| Keywords | C9 Deficiency / Genetic Analysis / RFLPs / Southern Transfer / 構造遺伝子 / RFLP / サザン・ブロッティング |
|---|
| Research Abstract |
High incidence of C9 deficiency in Japan has been clearified by Inai et al. The mechanisms of the defective synthesis of C9 observed in the C9 deficiency have not been resolved yet. We have studied RFLPs of DNA obtained from the cases with C9 deficiency.
1. DNA samples were prepared from leukocytes of 16 cases of C9 deficiency including two cases with Meningococcal meningitis.
2. DNA samples digested with the restriction endonucleases (EcoRI, BflII, HindIII, PstI, BamHI, etc.) were subjected to agarose-gel electrophoresis and alkali denaturation. The samples were transferred to the nylon filters and hybridized with the ^<32>P-labeled C9 cDNA probe. The banding patterns of homozygous C9-deficient subjects were similar to those of the normal subjects. These data exclude the possibility that the defective synthesis of C9 found in C9 deficiency may be due to a major deletion or rearrangement within the C9 gene.
3. We raised 16 monoclonal antibodies to C9. Fourteen antibodies including X195 bound to NH_2 terminal (C9a) fragments, and X197 bound to COOH terminal (C9b) fragments obtained by the cleavage of C9 with <alpha>-thrombin or trypsin. P40 recognized the epitope in the C9b obtained by <alpha>-thrombin cleavage but did not react with the NH_2-terminal or COOH-terminal fragments obtained by trypsincleavage. These antibodies may be available in the sensitive assay for C9 protein using ELISA.
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