| Project/Area Number |
61580218
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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 |
分子遺伝学・分子生理学
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| Research Institution | FACULTY OF SCIENCE, UNIVERSITY OF TOKYO |
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Principal Investigator |
IBA Hideo FACULTY OF SCIENCE, UNIVERSITY OF TOKYO, 理学部, 助教授 (60111449)
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| Co-Investigator(Kenkyū-buntansha) |
OKADA Yoshimi FACULTY OF SCIENCE, UNIVERSITY OF TOKYO, 理学部, 教授 (30011703)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1987: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1986: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | Cell cycle / Fibroblast / Anti-sense RNA / v-fos gene / c-fos gene / 細胞周期 / 線維芽細胞 / マイクロインジェクション / 増殖因子 |
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| Research Abstract |
In this research project, we intended to analyze the function of the c-fos gene that is transiently expressed upon external growth stimulations. For this purpose, we have tried to specifically inhibit the c-fos gene expression using anti-sense RNA or anti-fos antibody. Following materials or cell systems have been prepared for the project.
1. Isolation of v-fos transformed rat cell lines.
2. Preparation of a large amount (100 ug) og anti-sense RNA for v-or c-fos gene using SP6 RNA polymerase reaction in vitro.
3. Construction of retrovirus vectors which encode v-fos or c-fos (the vector can express v- or c-fos gene in CEF quite efficiently)
4. Construction of retrovirus vectors which express v-fos or c-fos anti-sensa RNA in CEF.
5. Cloning and nucleotide sequence analysis of chicken c-fos gene.
6. Preparation of anti-fos antibody which can immunoprecipitate v-fos, human, chicken and mouse c-fos gene products.
We have tried to screen the anti-sense RNA which has most inhibitory effect on v-fos or c-fos gene expression, by microinjection of antisense RNA or anti-sense RNA producing virus vectors. At present, we have find no effective anti-sense RNA and some residual expression of fos was always detectable. We are now trying to microinject anti-fos antibody into the cells for the specific and complete inhibition of the function of c-fos gene product.
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