Objective: To clone different domains of c-Fos in an eukaryotic vector harboring FLAG tag and to characterize their functions. Methods: Three different domains DNA were amplified by PCR method, and then were inserted into the eukaryotic expression vector FLAG-pcDNA3.0. The recombinant expression vectors were tran⁃siently transfected into 293T cell lines. The interaction between HA-tagged c-Jun and different FLAG-tagged c-Fos domains were demonstrated by co-immunoprecipitation. Results: The sequences of the cloned FLAG-tagged c-Fos domains were confirmed by DNA sequencing. The expression of the c-Fos domains was detected out by Western blot after collecting the cell lysate. Co-immunoprecipitation assays showed that c-Jun could interact specif⁃ically with the bZIP domain of c-Fos. Conclusion: Different domains of c-Fos were successfully constructed, which laid the foundation for further investigating the proteins that interact with the c-Fos.% 目的：克隆c-Fos蛋白不同结构域基因片段,构建其含FLAG标签的真核表达载体,并鉴定其功能.方法：用PCR方法扩增获得c-Fos不同结构域片段基因,定向克隆至本实验室保存的FLAG-pcDNA3.0载体中,瞬时转染293T细胞进行表达,并以本实验室保存的HA-c-Jun与FLAG-c-Fos不同结构域片段免疫共沉淀鉴定其相互作用区域,以佐证所构建的c-Fos结构域克隆的正确性.结果：测序结果表明c-Fos不同结构域序列正确,Western印迹显示可以在真核细胞中获得表达,并且免疫共沉淀结果显示c-Jun可以与c-Fos的bZIP区域特异性结合.结论：构建并表达了c-Fos不同结构域片段,为进一步研究与c-Fos相互作用的蛋白及其区域定位奠定了基础.
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