| Project/Area Number |
11460156
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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 | 一般 |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | Hiroshima University |
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Principal Investigator |
MORIKAWA Hiromichi (2000-2001) Hiroshima Univ., Graduate School of Science, Professor, 大学院・理学研究科, 教授 (00089129)
五島 直樹 (1999) 広島大学, 理学部, 助教授 (70215482)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Misa Hiroshima Univ., Graduate School of Science, Research associate, 大学院・理学研究科, 助手 (10294513)
木須 康智 広島大学, 理学部, 助手 (20304397)
森川 弘道 広島大学, 理学部, 教授 (00089129)
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| Project Period (FY) |
1999 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥11,100,000 (Direct Cost: ¥11,100,000)
Fiscal Year 2001: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1999: ¥4,100,000 (Direct Cost: ¥4,100,000)
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| Keywords | Matrix attachment region (MAR) / scaffold attachment region (SAR) / tobacco / gene delivery / green fluorescent protein (GFP) / integration / higher order structure of DNA / trangene / MAR (matrix attachment region) / SAR (nuclear scaffold attachment region) / パーティクルガン / 外来遺伝子 / 形質転換 / 核骨格結合部位(S / MAR) / タバコ培養細胞 / GFP (green fluorescent protein) / ゲノム DNA / 植物 / トランスジェニック植物 / PCR |
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| Research Abstract |
A 507-bp sequence (designated TJ1) that has been clone from a transgene locus of tobacco cell, and found to be a nuclear matrix / scaffold attachment region (MAR/SAR) was flanked to the 5' and 3' side of the expression cassette, in four different orientations, of the green fluorescent protein (GFP), a non-selective marker, gene. In each of these expression cassette, the GFP gene was under the control of cauliflower mosaic virus 35 S promoter and nopaline synthase polyadenlyation signal. These constructs were introduced separately into tobacco cells in order to investigate the effect of the MAR on the integration frequency of the transgene into the host genome. The presence of the MAR sequences in the cassette appeared to increase the transformant yield more than 2 times as compared with the control construct lacking the MAR sequences. Since there is no selection pressure to obtain those transformants in the present study, it is conceivable that MAR-based enhancement of the gene expression is not involved in the present result of the transformant yield. Therefore, the observed transformant yield should purely reflect the frequency of the integration of the transgene into the host genome. It is thus concluded that TJ1 MAR increases the integration frequency more than 2 times over the control. No significant effect of the orientations of the MAR sequence in the transformant yield was observed. Our findings provide direct evidence for the first time that the MAR cloned from the transgene locus stimulates the integration of transgene into the eukaryotic genome
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