Objective　To construct the prokaryotic expression vector for human Nm23-H1 genes and purify its fusion protein and study its activity.　Methods　Recombinant plasmid pET28a-Nm23-H1 was transformed into E.coli BL21 (DE3) and its expression was induced and identified with IPTG. The expressed Nm23-H1 fusion protein was purified by affinity chromatographic with Ni

2+

column and identified by SDS-PAGE and Western blotting. Immunogenicity and activity of Nm23-H1 fusion protein were analyzed by RP-HPLC and EMSA.　Results　The prokaryotic expression vector of pET2

8a-Nm23-H1 could express the Nm23-H1 fusion protein in E.coli BL21 (DE3). The Nm23-H1 fusion protein with NDPK activity could be purified by affinity chromatographic with Ni

2+

column. However

the AP endonuclease activity of Nm23-H1 protein was too weak to be detected.　Conclusion　Nm23-H1 fusion protein is highly expressed in E.coli BL21 (DE3) and can be purified，which is of great significance for elucidating the biologic characteristics of Nm23-H1 genes.