Objective To construct an eukaryotic expression vector and identify the integration of nasopharyngeal carcinoma-derived oncogene latent membrane protein 1 (N-LMP1) and CR2 gene in porcine embryonic fibroblasts, and to provide a basis for construction of EBV-infection associated swine nasopharyngeal carcinoma model.Methods ED-L2 and N-LMP1 were synthesized directly. CR2 was amplified by RT-PCR from human B lymphocytes. The three fragments mentioned above were subcloned one by one into the eukaryotic expression vector pN1, and then the vector was transfected into porcine embryonic fibroblasts using the liposome reagent, according to the manufacturer's protocol. The cells were selected with G418 antibiotic and identified by PCR amplification. Results N-LMP1 and CR2 epithelium-specific expression vector was successfully constructed and integrated into the genome of the porcine fibroblasts, and clone cells integrating the N-LMP1 and the CR2 genes were obtained. Conclusions The porcine fibroblast clones integrating N-LMP1 and CR2 are obtained and they should be of great value for the construction of N-LMP1 and CR2 transgenic swine via cell nuclear transfer.