| Project/Area Number |
10556071
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Applied veterinary science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
INABA Mutsumi Graduate School of Agricultural and Life Sciences, The University of Tokyo Associate Professor, 大学院・農学生命科学研究科, 助教授 (00183179)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAKUWA Yuichi Tokyo Women's Medical University Medical School, Professor, 医学部, 教授 (40113740)
YAMAMOTO Masayuki Tsukuba University, TARA Center, Professor, 基礎医学系先端学際領域研究センター, 教授 (50166823)
ONO Ken-ichiro Graduate School of Agricultural and Life Sciences, The University of Tokyo Professor, 大学院・農学生命科学研究科, 教授 (50111480)
KANEMAKI Misao Japan Live Stock Association, Head of Genetic Analysis Division, 家畜改良技術研究所, 遺伝検査部長(研究職)
MATSUKI Naoaki Graduate School of Agricultural and Life Sciences, The University of Tokyo Instructor, 大学院・農学生命科学研究科, 助手 (40251417)
高橋 迪雄 株式会社 味の素, 理事 (30011943)
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| Project Period (FY) |
1998 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1999: ¥3,500,000 (Direct Cost: ¥3,500,000)
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| Keywords | Gene therapy / transcription factors / GATA-1 / Promoters / Enhancers / Band 3 / Inberited disorders / Hematopoiesis / 赤血球 / 家畜 |
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| Research Abstract |
Reporter plasmid constructs were created in pSEAP vectors in which the reporter followed the transcription activating element derived from 5' upstream sequence of GATA-1 gene, including erythroid-specific promoter and exon 1, and intron 1. When transfected into K562 cells but not COS7 or CHO cells, activation of the reporter gene expression was observed. Thus, the region involving 5' upstream and intron 1 sequences was demonstrated to possess activity in enhancing gene transcription.
Retroviral vector containing the 5' upstream and intron 1 sequence from GATA-1 gene fused to normal bovine band 3 cDNA (pLNCX2bebWT) was prepared and several clones of K562 cells (K562bebWT) were isolated by G418 selection. Immunofluorescence staining showed that band 3 proteins localized to the plasma membrane, whereas the mutant construct (R664X) exhibited no significant signals at the cell surface. K562bebWT showed DIDS-sensitive chloride transport, demonstrating the protein have normal function. Introduction and expression of band 3 gene into bone marrow cells from affected animals are under investigation.
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