| Project/Area Number |
63860012
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Research Field |
応用生物化学・栄養化学
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| Research Institution | Tohoku University |
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Principal Investigator |
MIZUNO Shigeki Faculty of Agriculture, Tohoku University, Professor, 農学部, 教授 (90112903)
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| Co-Investigator(Kenkyū-buntansha) |
YUKI Atsushi Research Institute of Life Science, Snow Brand Milk Products Co. LTD. Chief Inve, 生物科学研究所, 主査
HARATA Masahiko Faculty of Agriculture, Tohoku University, Assistant, 農学部, 助手 (70218642)
結城 淳 雪印乳業(株), 生物科学研究所, 主査
西森 克彦 東北大学, 農学部, 助教授 (10164609)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥8,700,000 (Direct Cost: ¥8,700,000)
Fiscal Year 1990: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1989: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1988: ¥5,400,000 (Direct Cost: ¥5,400,000)
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| Keywords | sex chromosome markers / DNA-based sexing / W chromosome / glucocorticoid / glucocorticoid receptor / transgenic mice / グルココルチコイト レセプタ- / 染色体単離法 / MSB-1細胞(ニワトリの) / L細胞(マウスの) / TaqI 80bp反復単位(ニワトリの) / 同調培養 / pSV_2Neo / pRSVNeo / メチルセルロ-ス培地 / ニワトリのMAB1細胞 / cDNAライブラリー / チトクロームpー450c17 / ニワトリのZ染色体 / E1A遺伝子 / マイコフェノール酸 |
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| Research Abstract |
1) Finding of sex chromosome marker probes (by Mizuno and Harata) : XhoI-0.7kb and EcoRI-1.2kb fragments were cloned from the female chicken genomic DNA. These were repeating units of the W chromosome-specific XhoI- and EcoRI-repetitive DNA families, respectively. Slot blot and in situ hydridization demonstrated that these two clones were reliable probes for DNA-based sexing and to identify W chromosome in a nucleus and a metaphase chromosome set. Similar W chromosome-specific probes were also obtained from turkey and pheasant.
2) Inducible expression of introduced genes in animal cells (by Mizuno and Harata) : Co-transfection of pMMTV-LTR-cat and pMMGR followed by dexamethasone treatment induced CAT gene expression. The glucocortiocoid receptor gene expression from pMMGR was essential for this process in HeLa cells and chicken embryonic fibroblasts. This strategy was applied to inducible expression of nuclear lamina A/C cDNAs.
3) Stability of foreign genes in transgenic mice (by Yuki) : Transgenic mice which received pSV2-gpt-gElA sequences were mated and offsprings were analyzed for the presence of foreign DNA sequences. It was found that the foreign sequences integrated as head-to-tail concatamers were stably inherited but those integrated as head-to-head or tail-to-tail orientations were unstable and lost rather rapidly from the genome.
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