ANALYSIS OF THE FOS GENE FUNCTION IN THE CELL CYCLE.

在本研究项目中，我们打算分析c-fos基因的功能，这是瞬时表达的外部生长刺激。为此，我们尝试用反义RNA或抗fos抗体特异性地抑制c-fos基因的表达。以下材料或电池系统已准备用于该项目。v-fos转化大鼠细胞系的分离.体外用SP 6 RNA聚合酶反应制备大量（100 μ g）v-fos和c-fos反义RNA. 4.编码v-fos或c-fos基因的逆转录病毒载体的构建（该载体能在CEF中高效表达v-fos或c-fos基因）。在CEF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.制备能免疫沉淀人、鸡和小鼠v-fos基因产物的抗fos抗体，通过显微注射反义RNA或反义RNA产生病毒载体，筛选对v-fos或c-fos基因表达抑制作用最强的反义RNA。目前尚未发现有效的反义RNA，且常可检测到fos的残余表达。我们正在尝试将抗fos抗体注射到细胞内，以特异性地、完全地抑制c-fos基因产物的功能。

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.