Construction of eukaryotic expression vector for human prostate stem cell antigen and its expression

Objective To construct the eukaryotic expression plasmid of human prostate stem cell antigen(PSCA)containing a signal peptide and a FLAG tag and to detect its expression in eukaryotic 293T cells.Methods SIG-FLAG and human PSCA gene was constructed by PCR and inserted into a eukaryotic expression vector pIRES-neo.The recombinant plasmid pIRES-neo-sig-FLAG-PSCA was tansfected to the 293T cells,and its expression was detected by FACS,IMF and RT-PCR,respectively.Results The gene sequence of SIG-FLAG and human PSCA amplified by PCR was correct.Enzyme digestion analysis showed that the recombinant plasmid pIRES-neo-sig-FLAG-PSCA was successfully constructed and expressed in 293T cells.Conclusion The recombinant plasmid pIRES-neo-sig-FLAG-PSCA,we constructed,can be effectively expressed in 293T cells,which may lay a foundation for the construction of human PSCA-transfected cell line in the future.