| Project/Area Number |
07671220
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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 |
Hematology
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| Research Institution | Jichi Medical School |
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Principal Investigator |
KAJII Eiji Jichi Medical School, Dept.of Medicine, Professor, 医学部, 教授 (40204391)
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| Co-Investigator(Kenkyū-buntansha) |
IWAMOTO Sadahiko Jichi Medical School, Dept.of Medicine, Assis.Proff., 医学部, 助教授 (10232711)
土田 修一 自治医科大学, 医学部, 講師 (20217326)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1996: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1995: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Red blood cell / Differentiation / Blood group system / Rh blood group / Duffy blood group / Splicing isoform / Gene / mRNA / Rh抗原 / Duffy抗原 / 遺伝子導入 / Rh50糖蛋白 / Rh変異型 / mRNA / Rh蛋白 / Duffy蛋白 / スプライシング |
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| Research Abstract |
Gene structure analyzes of the Rh blood group system revealed that both the RHD and RHCE genes consist of 10 exons and 9 introns that a 8kb-spacer region connects these two genes. By a systemic analysis of Rh-related cDNA isoforms based on reverse transcription-polymerase chain reaction, 11 and 5 truncated isoforms of the RhCE and RhD cDNAs, respectively. Studying on Rh-related gene expression during human erythroid differentiation exhibited the existence of isoforms inducible during erythroid maturation.
We performed epitope analysis of Rh antigen by constructing retroviral gene coding 6 normal and chimera Rh cDNAs. The cDNAs were introduced into KU812E cells. The results indicated that the Rh epitopes were not constructed only with short polymorphic exofacial peptide loops but with other peptide fragments and other membrane components. Co-expression studies of RH50 and RHD or RHCE gene in non-erythroid cells, 293, did not show any Rh antigens and suggested that at least a second co-expressing factor was needed to express RhD or RhCE antigens on the plasma membrane. The gene analysis of a propositus with Rhnull phenotype confirmed the Rh50 glycoprotein as one of the co-expressing factors.
We found a novel first exon and spliced form of Duffy mRNA that is the predominant transcript in both erythroid and postcapillary venule endothelium. We also identified two inverse GATA motifs in the 600 bp 5' flanking region. The proximal GATA was positionated downstream from the start position of endothelium and upstream from that of erythroid. Gel shift assay showed the proximal GATA gene is the target sequence of GATA-1. Deletion mutagenesis study revealed that this GATA motif represent the erythroid regulatory core region for Duffy gene.
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