Objective:To express and characterize human tumor protein D52 (hTPD52) protein using the proper prokaryotic expression vector in E.coli. Methods:The hTPD52 gene was amplified by RT-PCR from MCF-7 cell line,and cloned into vector pMD18-T. After sequenced,the correct fragment of hTPD52 was inserted into three different prokaryotic expression vectors,which were then transformed into E.coli.BL21(DE3),respectively. The optimal expression condition was selected. After induced with IPTG,the recombinant protein was purified with His Trap affinity chromatography,and characterized by Dot blot and Western blot. Results:The hTPD52 with isoform 3 was amplified and sequenced correctly,and the recombinant expression plasmid was constructed successfully. SDS-PAGE analysis showed that the molecular weight of hTPD52-Trx fusion protein was about 44ku. The purified protein could be used as antigen to detect the specific anti-hTPD52 antibody by Dot blot and Western blot. Conclusion:The recombinant fusion protein of hTPD52 could be expressed with high performance in E.coli,and the antigenicity of hTPD52 protein is retained for the research in the future.