Objective To investigate the expression of rpl15 in zebrafish. Methods The NCBI and Ensemble databases were used to analyze the co-collinearity of the rpl15 gene and the sequence similarity of rpl15 protein. MEGA X software was used to align rpl15 in different organizations. The mRNA expression of rpl15 was detected in whole embryos by in situ hybridization with an rpl15 antisense RNA probe, and the expression of rpl15 protein in 24, 48, and 72 hpf zebrafish embryos was detected by western blotting. Results The rpl15 gene is relatively conserved during evolution. A pCS2⁺ -rpl15 recombinant plasmid was constructed, and in situ hybridization result showed that at 24 hpf, rpl15 was widely expressed in most cells of zebrafish. At 48 hpf, rpl15 expression was highest in the central nervous system and it was also present in the pancreas. At 72 hpf, rpl15 expression was still present in the posterior brain, as well as the pancreas, liver, and intestine. The relative protein expressions of rpl15 in 24, 48, and 72 hpf zebrafish embryos increased with increasing time. Conclusions Changes in the expression sites and expression levels of rpl15 in zebrafish during development were systematically observed, which provide the experimental basis for establishing a model of rpl15 deletion in zebrafish.