Abstract: Objective To construct eukaryotic expression vector of human hepatocyte nuclear factor (HNF）-4α gene and identify its expression in vitro. MethodsThe total RNA was isolated from the liver tissues of two patients with hepatocellular carcinoma (HCC） who had underwent surgical operation, and HNF4α cDNA was synthesized by reverse transcription and amplified with the existence of specific primers. Then, the HNF4α fragment was directionally linked to pcDNA3.1 positive-eukaryotic expression vector. After antibiotic screening, the sequence analysis was conducted. The vector was transfected into HepG2 cells, and the expression of target protein after 48-hour incubation was detected by Western blot. Results Enzyme digestion and sequencing illustrated that pcDNA3.1-4α positive-eukaryotic expression vector was successfully constructed, and the results of Western blot revealed that obvious band of fusion protein was present at 53 kDa position. Conclusion The eukaryotic expression vector of HNF4α gene is successfully constructed, and it works well in HepG2 cells in vitro, which is good for further study of HNF4α protein.

Key words: HepG2 cells, Hepatocyte nuclear factor-4α, Eukaryotic expression, Cytokines