ANALYSIS OF THE FOS GENE FUNCTION IN THE CELL CYCLE.

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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.