Abstract: Objective: To amplify human prostate stem cell antigen（PSCA） genes and construct a eukaryotic expression plasmid to express human PSCA. Methods: The genes of PSCA were amplified by RT PCR from human prostate cancer cell line LNCaP,and then cloned into pIRES neo-eukaryotic expression vactor to construct the pIRES neo PSCA His plasmid.Through the mediation of lipofectamine,the plasmid was transfected into B16 cells,and then the transfected B16 cells were preliminarily selected by G418.The transcription of PSCA mRNA and the expression of protein in B16 cells were detected by RT PCR and flow cytometry and immunofluorescence assay respectively. Results: The 372bp target PSCA genes identical with the sequence of GenBank were obtained.The plasmid was successfully constructed and transfected into B16 cells.The eukaryotic expression was approved. Conclusion: The successful construction of the eukaryotic expression plasmid pIRES neo PSCA His will contribute to the research of PSCA in the immunotherapy of prostate cancer.