Objective To construct recombinant lentivirus vector of rat Cdh1 gene. Methods The artificially synthesized full lenth DNA of rat Cdh1 gene was inserted in the pGC-FU vector, which was isolated by restriction enzyme digest with Age I. The positive recombinant was identified by PCR analysis and sequence analysis. Six culture dishes of 293T cell were randomly allocated into intervention group and control group. Expression of GFP protein was detected by fluorescence microscope and Westem blot in pGC-FU-Cdh1 transfected 293 T cells. Lentivirus pacaging, concentration and titering were done. Results The PCR analysis and sequence analysis demonstrated that the size and position of Cdh1 gene insertion were consistent with the design. Westem blot analysis showed specific expression of external fusion protein of Cdh1-GFP in pGC-FU-Cdh1 transfected 293 T cells(n=3), and have no expression in control group (P=0.014). Lentivirus titering by Real-time quantitative PCR was 2E+8 TU/ml. Conclusions The effective recombinant lentivirus vector of rat Cdh1 gene was successfully constructed and may serve as the basis for the further study of Cdh1 gene function.