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In order to study the structure and function of DLX5 gene in sika deer (Cervus nippon), and to further explore the underlying ossification mechanism of sika antler velvet，DLX5 gene of sika deer was cloned by RT-PCR. The cDNA sequence containing all coding regions was obtained. The amino acid sequence of the gene was analyzed by using bioinformatics method and the phylogenetic tree was constructed. The signaling pathway was analyzed by KEGG enrichment, and finally real-time RT-PCR was used to detect the expression of DLX5 gene in different growth periods of deer antler velvet. The results showed that the coding region of DLX5 gene in sika deer was 870 bp, and a total of 289 amino acids were encoded. DLX5 protein was a soluble, unstable protein with two conserved domains and was mainly localized to the nucleus. Sika deer DLX5 protein had a high similarity with the amino acid sequence of DLX5 proteins derived from the different species, which was relatively conservative. Irregular curl accounted for the largest proportion in the secondary structure of DLX5 protein, and followed by α -helix and extended chain. Irregular curl, α -helix and extended chain was 78. 89%, 16. 96% and 4. 15%, respectively. The main pathways of DLX5 gene were TGFβ signaling pathway, MAPK signaling pathway and Wnt signaling pathway. The real-time RT-PCR results showed that the expression level of the gene was significantly increased in the late stage of deer antler growth m(trident velvet), and the upregulated expression suggested that it played an important role in the process of deer antler ossification, which may be a candidate gene for deer antler ossification. [ABSTRACT FROM AUTHOR]