The invention discloses an infectious cDNA clone of canine distemper virus CDV‑3 strain, a construction method and application thereof. In the present invention, the canine distemper virus CDV-3 strain is used as a template, and the full length of CDV-3 is divided into five fragments for RT-PCR amplification. After enzyme digestion and splicing, the 5 fragments were sequentially inserted into the eukaryotic vector, and the sequences of hammerhead ribozyme and hepatitis D ribozyme were added to the head of F1 and the end of F5, respectively, to obtain the infection of canine distemper virus CDV‑3 strain sex cDNA clones. Then, construct three helper plasmids expressing CDV-3N, P, L protein. The infectious cDNA clone of canine distemper virus CDV‑3 strain and three helper plasmids were co-transfected into 293T cells to obtain rescued recombinant CDV‑3 virus (rCDV‑3). The study found that rCDV‑3 viral titers were obtained up to 10 7.667 TCID 50 / mL, 10 times higher than wtCDV‑3. After infection of Vero cells with rCDV‑3, the highest virus titer was rapidly reached at 36 hours after infection, while the virus content of wtCDV‑3 reached the highest at 72 hours after infection. The proposal of the invention lays a foundation for the development of a novel vaccine against canine distemper virus and the study of its pathogenic mechanism.