| Project/Area Number |
04680045
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory animal science
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| Research Institution | Tokai University, School of High-Technology for Human Welfare. |
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Principal Investigator |
HYODO Masao Tokai University, School of High-Technology for Human Welfare. Professor., 開発工学部, 教授 (60096253)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1993: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Medaka / Embryonic stem cell / Chimera / Hatching enzyme / Feeder cell / ジーントラップ法 / トランスジェニック / ジーントラップ / 培養細胞 / lacZ遺伝子 |
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| Research Abstract |
Establishing an experimental system for gene-targeting or gene-trapping in transgenic fish requires a method to manipulate the embryonic stem(ES)cells. As an initial step, we have tried to produce chimeras in medaka(Oryzias latipes)by transplantation of the ES cells from embryos at an early stage of development. Hard egg chorion had prevented delivery of the donor cells to recipient embryos, but partial digestion of the chorion by a crude fraction of the hatching enzyme enabled insertion of the microcapillary tip into the embryonic cell layr. A wild-type black-pigmented strain was used as a donor and the cells were microinjected into the recipient embryo of a mutant orange-colored strain, both at the mid-blastula stage. In 9 successful transplants, 4 fishes showed clear chimeric body color. And the germ-line transmission of the wild-type cells was confirmed in 5(55%)of the transplants by production of the black-pigmented progeny when they were mated with the orange colored mutants. The results indicate that a method used in this study was suited for embryonic cell manipulation in medaka.
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