Mammal Toll-like receptor 5 (TLR5) can directly recognize bacterial flagellin, initiate the inflammatory signaling cascades and trigger body immune system to clear the "non-self" substances. In teleosts, TLR5 has presented more complexes not only in increasing the molecular types, but also in elevating the functional diversity. In this study, we identified two TLR5 family members in Schizothorax prenanti, named as spTLR5-1 and spTLR5-2. The complete coding sequence (CDS) of spTLR5-1 is 2622 bp, encoding 873 amino acids, while the complete CDS of spTLR5-2 is 2640 bp, encoding 879 amino acids. Phylogenetic analysis showed that spTLR5-1 and spTLR5-2 were clustered to the TLR5 of schizothorax richardsonii and Cyprinus carpio respectively. The 3D structure analysis exhibited that the