Abstract: In order to provide the donor cells for nuclear transfer, in this study, the eukaryotic expression vector PEGFP-SRA-Ipr1 was constructed, the bovine fetal fibroblast cells were isolated in vitro and the cells with PEGFP-SRA-Ipr1 were transfected. Ipr1 gene and SR-A promoter were successfully cloned and then used to construct the macrophage-specific eutherin expression vector- Bovine fetal fibroblast cells from ear were cultured in vitro by tissue explantation, passaged and purified. Then the eutherin expression vector PEGFP-SRA-Ipr1 was transfected into the cells at the passages of 4th to 10th by using electroporation. The fluorescence was observed after 24 h. 600 μg·mL^-1 G418 was added into the culture medium after 48 h, which lasted 1 week. After that, the concentration of G418 decreased to 300 μg·mL^-1 for screening. Then the monoclones were picked and cultured for amplification The positive cells were determined via PCR and flow cytometry. GFP was observed 24 h after transfection. The bovine fetal fibroblast cells permanently carrying PEGFP-SRA-Ipr1 were obtained after the screening of G418. The interested fragment was confirmed to be correct by PCR and the positive cells had the normal karyotype, suggesting that the interested gene was successfully integrated into the genome and it failed to disrupt the stability of the genome. In conclusion, the obtained cells could be utilized as the donor cells for research of transgenic cloned cattle by nuclear transfer.