As one of the important members of interleukin, interleukin-11(IL-11) plays a pivotal role in many biological processes, including wound healing, cancer cells migrating, immunity regulating, and so on. To study the transcription regulatory mechanism of il-11b, the 3 000 bp promoter of an interleukin-11b (il-11b) in zebrafish (Danio rerio) was found through bioinformatics analysis, and we obtained 2 908 bp, 2 000 bp, 1 000 bp and 500 bp different fragments after deletion processing. These promoter sequences were amplified through PCR method and reconstructed into pGL3-enhancer empty plasmid to form reporter gene vectors. We used dual-luciferase reporter assay system to detect the activity of these promoters in HEK293T cell line. Bioinformatics results indicated there are about 3 promoter regions and 7 CpG islands in the sequence of il-11b promoter, and 60 binding sites including Adf-1, Krox-24, Sp1, CACCC-box, Egr-1, ICSBP, c-Jun, COUP, GR, RAP1, NF-kappaB, REV-Erbalpha, ER, RAR-alpha1, RXR-beta, SRF, Oct-1, Oct-2, Pit-1a, GCN4, HNF-1, TBP, C/EBPalpha, HNF-1C, Oct-2.1, USF, etc. The fragment sizes of all recombinant plasmids were consistent by double enzyme digestion, and the sequences were correct. Compared to control group, the results of luciferase showed the activity of pGL3-il-11b, pGL3-il-11b-Δ1, pGL3-il-11b-Δ2, pGL3-il-11b-Δ3, pGL3-il-11b-Δ4, pGL3-il-11b-Δ5 were about 1.94, 14.92, 5.33, 9.11, 0.75, 1.32 folds, respectively. These results may provide a tool to investigate the immune signals in which cytokines are involved.