Abstract: Objective To clone human NR2B gene, construct its eukaryotic expression vector, and temporarily express it in CHO cells. Methods Human NR2B gene was amplified by RT-PCR and then inserted into eukaryotic vector pcDNA3.1. The recombinant plasmid was transfected into CHO cells. The expression of the target molecule was identified by RT-PCR, Western blotting, indirect immanofluorescent staining and the apoptosis was detected by flow cytometry. Results The NR2B gene was obtained; after transfection, NR2B was successfully expressed in CHO cells, and the expression of NR2B did not induce the apoptosis of CHO cells. Conclusion Human NR2B gene has been successfully cloned and expressed in CHO cells via constructing its eukaryotic expression vector.