Objective：To construct the recombinant eukaryote expression vector containing Max dimerization protein 1（Mad1）gene and detect its effect on gastric cancer cell proliferation and migration. Methods： The Mad1 gene was cloned into pEGFP-N1 expression vector by recombining DNA technology. The recombinant vector was identified by restriction enzyme analysis and nucleotide sequence determination. The eukaryotic expression plasmid pEGFP-N1-Mad1 was transiently transfected into AGS cells. Expression of Mad1 gene and protein was identified by RT-PCR and Western blot，respectively. The location of Mad1 protein was detected by fluorescence microscope. The proliferation and migration of AGS cells were examined by CCK-8 and Transwell assay，respectively. Results：The Mad1 gene was successfully cloned to the eukaryote expression vector pEGFP-N1. Expression of Mad1 gene and protein was confirmed by RT-PCR and Western blot. After transfection，Mad1 could be detected in the nucleus of AGS cells. CCK-8 and Transwell experimental results showed that the proliferation and migration of pEGFP-N1-Mad1 transfected cells were deteriorated significantly compared to empty vector transfected AGS cells and normal AGS cells. Conclusion：The new recombinant expression vector pEGFP-N1-Mad1 was constructed and expressed successfully in AGS cells. Mad1 could inhibit the proliferation and migration of gastric cancer cells.