C-type lectin, which plays an important role in fish innate immunity, was cloned from tilapia and its functional characterization under low-temperature stress is reported. Its ORF is 453 bp, encoding 150 amino acids, and has a 5′UTR of 83 bp, a 3′UTR of 559 bp, and a poly (A) tail. The tilapia C-type lectin genomic DNA was acquired with a length of 5714 bp, containing six exons and five introns. Its promoter sequence was cloned and has a length of 2251 bp. The highest promoter activity occurs in the regulatory region (−900 bp to −450 bp). A hemagglutination assay of recombinant tilapia C-type lectin protein showed positive hemagglutination of rabbit and tilapia erythrocytes. RT-qPCR and western blot assays showed that its expression in the liver, spleen, and intestine were clearly affected by low-temperature stress. Thus, tilapia C-type lectin appear to be affected by abiotic stress, as well as by biological stress.