Objective: To investigate the construction of eukaryotic expressing vector containing human full-length Tip30 gene (pAAV-TIP30) and observe its expression in the transfected hepatocellular carcinoma (HCC) cell line HepG2. Methods: Using the mRNA extracted from normal human liver tissue as a template, Tip30 gene was amplified through reverse transcription polymerase chain reaction (RT-PCR) method. The PCR products were digested with BamH I and Xba I, and cloned into eukaryotic expressing vector pAAV-MCS. The recombinant plasmids were then transfected into the HepG2 cells and the expression of TIP30 protein in the host cells was detected by Western blot analysis. Results: Full-length TIP30 gene was correctly amplified by RT-PCR method. The identity of the cloning of TIP30 gene in pAAV-MCS was confirmed by restrictive enzyme digestion and sequencing. Western blot showed that the protein expression of TIP30 increased in the HepG2 cells, after they were transfected with the recombinant plasmids. Conclusion: Recombinant plasmid pAAV-Tip30 has been cloned and transfected into HepG2 cells successfully, which can provide a basis for the further investigation of the effects of TIP30 gene on HCC and the underlying mechanisms as well.