ANALYSIS OF THE FOS GENE FUNCTION IN THE CELL CYCLE.

细胞周期中 FOS 基因功能的分析。

• 批准号: 61580218
• 负责人: IBA Hideo
• 金额: $ 1.02万
• 依托单位: FACULTY OF SCIENCE, UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61580218/
• 关键词: 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

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.

在这个研究项目中，我们打算分析在外部生长刺激下瞬时表达的 c-fos 基因的功能。为此，我们尝试使用反义RNA或抗fos抗体特异性抑制c-fos基因表达。本项目已准备了以下材料或细胞系统： 1． v-fos转化大鼠细胞系的分离。2．利用体外SP6 RNA聚合酶反应制备大量(100ug)og v-或c-fos基因反义RNA。3．构建编码v-fos或c-fos的逆转录病毒载体（该载体可以在CEF中高效表达v-或c-fos基因）4。在CEF.5中表达v-fos或c-fos反义RNA的逆转录病毒载体的构建。鸡c-fos基因的克隆及核苷酸序列分析。6．制备抗fos抗体，可免疫沉淀v-fos、人、鸡、小鼠c-fos基因产物。我们尝试筛选对v-fos或c-fos基因表达最具抑制作用的反义RNA，通过显微注射反义RNA或产生反义RNA的病毒载体。目前，我们还没有发现有效的反义RNA，并且始终可以检测到一些fos的残留表达。我们现在正在尝试将抗fos抗体显微注射到细胞中，以特异性、完全抑制c-fos基因产物的功能。

项目成果

H. Iba, Y. Shindo, H. H. Nishina and T. Yoshida: "Transforming potential of the v-fos and c-fos genes carried by avian retrovirus vectors." Oncogene Research.in press. (1987)

H. Iba、Y. Shindo、H. H. Nishina 和 T. Yoshida：“禽类逆转录病毒载体携带的 v-fos 和 c-fos 基因的转化潜力。”

K. T. Fujiwara, K. Ashida, H. Nishina, H. Iba, N. MIyajima, M. Nishizawa, & S. Kawai: "The Chicken c-fos gene; c10ning and nucleotide sequence analysis" J. Virol. 61. 4012-4018 (1987)

K. T. Fujiwara、K. Ashida、H. Nishina、H. Iba、N. MIyajima、M. Nishizawa、

J,Kato: Mol.Cell.Biol.6. 4155-4160 (1986)

J，加藤：Mol.Cell.Biol.6。

H. Nakano, E. Kobayshi, T. Takahashi, T. Tamaki, Y. Kuzuu, and H. Iba: "Staurosporine infibits tyrosine-specific protein kinase activity of Rous sarcoma virus transforming protein p60." J. Antibiotics. 40. 706-708 (1987)

H. Nakano、E. Kobayshi、T. Takahashi、T. Tamaki、Y. Kuzuu 和 H. Iba：“Staurosporine 抑制劳斯肉瘤病毒转化蛋白 p60 的酪氨酸特异性蛋白激酶活性。”

R.Jove: J.Virsl.60. 840-848 (1986)

R.Jove：J.Virsl.60。