Abstract: In order to further understand the biological and cytological functions of buffalo DDR1 protein, for in this study, ProtParam, ProtScale, SOPMA, SWISS-MODEL, PSORT Ⅱ Predictio and NetPhos 3.1 online analysis softwares were used to bioinformatics analysis. Meanwhile, the effects of DDR1 gene on proliferation, apoptosis, gene expression and migration in buffalo breast epithelial cells were studied by gene interference method. Results showed that buffalo DDR1 protein was composed of 915 amino acids with a formula of C₄₅₃₈H₇₀₂₉N₁₂₆₁O₁₂₈₇S₄₃, molecular weight of 101 222.99 u, half-life of 30 h, and theoretical isoelectric point of 6.22, which was an acidic, unstable and soluble protein. The secondary structure of DDR1 protein consisted of 256 alpha helixes（27.98%）, 175 extended strands（19.13%）, 55 beta turns（6.01%）, and 429 random coils（46.89%）. Subcells were localized in the endoplasmic reticulum（44.4%）, golgi apparatus（33.3%）, and plasma membrane（22.2%）. The DDR1 protein contained a total of 65 phosphorylation sites, including 35 serine phosphorylation sites, 18 threonine phosphorylation sites, and 12 tyrosine phosphorylation sites. The optimal interference fragment of the DDR1 gene was siDDR1-1, and the optimal interference time was 48 hours. At 72 hours of DDR1 gene interference, the proliferation of buffalo mammary epithelial cells was significantly inhibited（P＜0.01）. DDR1 gene interference significantly up-regulated the expression of apoptosis-related genes（BCL-2P＜0.01, XIAPP＜0.05, and TP53P＜0.01） and proliferation-related gene CCND1,and down-regulated the expression of apoptotic-related genes（CASPASE3P＜0.05 and CCDB1P＞0.05）, but had no significant effects on apoptosis（P＞0.05）. In addition, cell migration in the interfering group was significantly reduced（P＜0.01）. The results indicated that DDR1 gene interference could affect the proliferation and migration of buffalo mammary epithelial cells.