| Project/Area Number |
11356008
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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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 animal science
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| Research Institution | AZABU UNIVERSITY |
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Principal Investigator |
TACHI Chikashi Azabu University, School of Vet Med., Professor, 獣医学部, 教授 (30011711)
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| Co-Investigator(Kenkyū-buntansha) |
YOKOYAMA Minesuke Mitsubishi Kasei Institute of Life Science, Reproductive Engineering Section and Exerimental Animal Center, Chief, 生殖工学開発室, 室長
SATOH eimei Tohoku University, Graduate School, Professor, 大学院・農学研究科, 教授 (80093243)
KASHIWAZAKI Naomi Azabu University, School of Vet Med, Associate professor, 獣医学部, 助教授 (90298232)
HACHISU Matsuyuki Shibayagi Co., Ltd., Managing Director, 代表取締役
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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 |
¥36,100,000 (Direct Cost: ¥33,700,000、Indirect Cost: ¥2,400,000)
Fiscal Year 2001: ¥10,400,000 (Direct Cost: ¥8,000,000、Indirect Cost: ¥2,400,000)
Fiscal Year 2000: ¥12,300,000 (Direct Cost: ¥12,300,000)
Fiscal Year 1999: ¥13,400,000 (Direct Cost: ¥13,400,000)
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| Keywords | goart / transgenic / transgenic technologies / telomere length / myostatin / Sry / antisense / riobzyme / 精子の凍結保存 / 体外受精 / 卵成熟 / DAX1 / myostatin / HMGI-C / トランスジェニックマウス |
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| Research Abstract |
Production of transgenic domestic animals in aitiodactyls with relatively small and facility of handling has become commercial feasibility in US and European countries. On the other hand, reliable methodology for the production of Tg animals has not yet been sufficiently worked out in Japan. The aim of present project is to develop practical Tg technologies in goats. The following researches had been carried out ; establishment of cultured cell line (CPF 1) as a donor for nuclear transplantation, analysis of chromosome numbers and relationship between telomere length and generation numbers in CPF 1, assessment of cryopreservation for spermatozoa and establishement of IVM/IVF procedures in goats. Attempts for molecular cloning of genes and/or cDNAs coding for protein factors potentially useful in genetic engineering of goats as a model of artiodactyl farm animals has been made and yielded noteworthy successful outcomes. The cDNAs coding for myostatin which regulates development of skele
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tal muscles, and HMGI-C responsible for the control of body-size have successfully been cloned and sequence-analyzed. Two sex determining genes, SRY and DAX1 have also been cloned and their nucleotide sequences were determined. Expression vectors for antisense murine Sry gene were constructed and Tg mice were produced. As the promoter/enhancer complex which drives the antisense -Sry trnasgene, CAGGS construct was employed. Twelve Tg mice were obtained as the founders where the genotypes coincided with the sexual phenotypes. In F1 derived from the founders, preliminary PCR analysis of the genes yielded results which led us to suspect the possible presence, of phenotypic females. Further confirmatory experiments are currently in progress. With regard to the development of anti-mSry ribozymes, we designed several ribozyme. The effects of these ribozymes against the Sry mRNA expressed in vitro were examined and evidence was obtained that the hammerhead ribozyme and one riobzyme-tRNA complex were capable of effectively cleaving the Sry mRNA. Less
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