1990 Fiscal Year Final Research Report Summary
Introduction of Genes and Chromosomes into Animal Cells and Fertilized Eggs
| 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)
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| Project Period (FY) |
1988 – 1990
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| Keywords | sex chromosome markers / DNA-based sexing / W chromosome / glucocorticoid / glucocorticoid receptor / transgenic mice |
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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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