1999 Fiscal Year Final Research Report Summary
Generation of fish clone by nuclear transplantation of cultured cell in medaka
| Project/Area Number |
10559011
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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 |
広領域
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| Research Institution | NAGOYA UNIVERSITY |
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Principal Investigator |
WAKAMATSU Yuko Nagoya University, Bioscience Center, Associate Professor, 生物分子応答研究センター, 助教授 (20026800)
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| Co-Investigator(Kenkyū-buntansha) |
ARAKI Kazuo National Research Institute of Aquaculture, Chief Researcher, 水産庁養殖研究所・遺伝育種部, 主任研究官
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| Project Period (FY) |
1998 – 1999
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| Keywords | medaka / nuclear transplantation / cultured cells / fish clone |
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| Research Abstract |
Although nuclear transplantation is an important technique of modern biology, it has not been established yet in fish. The purpose of the present study is to develop techniques of fish cloning by nuclear transplantation of cultured fish cells in medaka. The study was conducted following steps.
1) Nuclear transplantation of embryonic cells into nonenucleated oocytes : We could obtain adults nuclear transplants with genetic markers from both of donor nuclei and recipient oocytes, but they were triploid and unfertile.
2) Nuclear transplantation of embryonic cells from transgenic fish carrying the medaka elongation factor 1 α-A promoter/GFP (EF-1 α-A/GFP) into nonenucleated oocytes : We could obtain adults nuclear transplants with GFP fluorescence, but they are triploid and unfertile.
3) In order to generate diploid nuclear transplants, oocyte nuclei were inactivated by 100 to 200 Gy of X-rays.
4) Nuclear transplantation of embryonic cells from two transgenic fish carrying EF-1 α-A/GFP and medaka β -actin promoter/GFP (β -act/GFP) into enucleated oocytes. We could obtain adults nuclear transplants with natural and GFP makers of donor nuclei, which were diploid and fertile. The transgenes were expressed faithfully to property of promoters used. The donor genetic markers were transmitted to subsequent generations in Mendelian fashion. Nuclear transplantation of cultured cells is in process.
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