Project/Area Number |
10670943
|
Research Category |
Grant-in-Aid for Scientific Research (C)
|
Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Hematology
|
Research Institution | Mie University |
Principal Investigator |
WADA Hideo Mie University, Faculty of Medicine, assistant, 医学部, 助手 (40158704)
|
Co-Investigator(Kenkyū-buntansha) |
渡辺 正人 三重大学, 医学部・附属病院, 助手 (30220924)
|
Project Period (FY) |
1998 – 1999
|
Project Status |
Completed (Fiscal Year 1999)
|
Budget Amount *help |
¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1999: ¥1,000,000 (Direct Cost: ¥1,000,000)
|
Keywords | ITP / SEREX / cDNA / specific antibody / sequence / UT-7 / Molecular biology / recombinant protein / 細み替え蛋白 / dipA / IgG抗体 / 遺伝子解析 / 遺伝子クローニング |
Research Abstract |
To identify the cause of specific antigen in idiopathic thrombocytopenic purpura (ITP), we directly analyzed the gene of specific antigen in ITP by high sensitivity of genetic cloning method ; serological identification of antigens by recombinant expression cloning (SEREX). We extracted the RNA from UT-7/thrombopoietin and made the cDNA. After expression of protein from the cDNA in E-Coli, 38 clones which were reacted to the serum of patients with ITP were obtained. Out of them, 9 clones were unknown and 19 were known by the analysis of sequence of gene. We focused human hepatitis delta antigen interacting protein A (dip A), because it strongly reacted to the ITP serum. By RT-PCR method, we detected the expression of dip A in all cell line. We examined the anti-dip A antibody in patients with ITP by western blot method. Sixty percent of ITP patients were positive for dip A antibody, but only 20% of healthy volunteers were positive. The possibility of point mutation in dip-A and role of anti-dip-A antibody in the onset of ITP should be studied furthermore.
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