Objective To explore the tetracycline-inducible expression system in zebrafish application strategy and build a transgenic zebrafish with tetracycline-inducible and liver-specific expression of green fluorescent protein. It is a fundamental work to construct conditional gene function studies and tissue-specific transgenic zebrafish disease models. Methods In this experiment pfabp10-rtTA was constructed with rtTA under the liver-specific promoter fabp10. pTRE-Tight-BI-AcGFP1 and pfabp10-rtTA were co-transfected into HeLa cells to confirm the doxycycline induction, and after that two plasmids were injected into zebrafish 1-cell stage embryos. After 30 μg/mL doxycycline incubation, the integrated individuals stably expressing GFP in the liver were selected. Results HeLa cells were transfected with pfabp10-rtTA and pTRE-Tight-BI-AcGFP1 plasmids and cultured with 1 μg/mL doxycycline, and take 0 μg/mL concentration of doxycycline as control. Western blot results showed that GFP expression was significantly higher in the experimental group than in the control group. The selected stable integrated zebrafish were cultured at a concentration of 30 μg/mL doxycycline conditions, with obvious GFP expression in the liver, while in the control group without doxycycline conditions, no obvious GFP expression was seen in the liver. Conclusions Our findings indicate that the use of the tetracycline-inducible expression system in liver of transgenic zebrafish lines could provide a good animal model tool for establishing conditional transgenic zebrafish disease models, and serve the research such as liver organogenesis and development of liver diseases.