| Project/Area Number |
10640601
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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 |
遺伝
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| Research Institution | Nagoya University |
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Principal Investigator |
HORI Hiroshi Nagoya University, Graduate School of Science, Division of Biological Sciences, Professor, 大学院・理学研究科, 教授 (60116663)
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| Co-Investigator(Kenkyū-buntansha) |
KOGA Akiko Nagoya University, Graduate School of Science, Division of Biological Sciences, Associate Professor, 大学院・理学研究科, 助教授 (80192574)
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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 |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | medaka / Oryzias latipes / transposable element / Tol-2 / tagging / トランスポゾン / 遺伝子改変系 / トランスポゼースmRNA / トランスポゼースcDNA / 人工mRNA |
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| Research Abstract |
We have identified a novel terminal-inverted-repeat class transposable element in the medaka fish Oryzias latipes. The element was designed Tol-2. Excision of Tol-2 during embryogenesis was detected by a PCR analysis of a genomic region containing a Tol-2 copy. Tol-2 is 4681 bp in length. Tol-2 contains 4 ORFs which have amino acid sequence similarity with Ac of maize, hobo of Drosophia and Tam3 of snapdragon, which are all active transposable elements. Tol-2 is thus a unique material for establishing a gene tagging system in fish. We developed a novel transient embryonic excision assay in which medaka fertilized eggs were coinjected with mRNA transcribed in vitro using the Tol-2 cDNA as a template and a plasmidDNA harboring a Tol-2 element inside the b-galactosidease gene. The Tol-2 element could be efficiently excised only when co-injected with the Tol-2 mRNA. The excision activity is measured efficiently by using the Tol-2 containing plasmid vector.
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