[Inhibitory effect of recombinant HMGN2 protein on human hepatitis B viral].

ABSTRACT

OBJECTIVE: To construct a prokaryotic expression recombinant for the expression of HMGN2 and to evaluate its antiviral activity against human hepatitis virus. METHODS: The extracellular region cDNA of HMGN2 was isolated and amplified by RT-PCR, and introduced to the prokaryotic expression vector pGEX-4T-1. HMGN2 protein was expressed under IPTG induction and purified by GST protein purification system, then identified by SDS-PAGE and Western blot. The cytotoxicity of fusion HMGN2 to HBV-transfected HepG2. 2.15 cell was evaluated with MTT assay. Different concentration of fusion HMGN2 was applied on the HepG2. 2.15 cell and the cell culture supernatants were harvested after 3 and 6 days treatment. The HBsAg and HBeAg in the supernatants were detected by ELISA and the HBV DNA was detected by RT-PCR. RESULTS: In the range of tested 1-100 microg/mL of HMGN2, no cytotoxicity to HepG2. 2.15 cells was detected by MTT assay. When incubated with HMGN2 at 15 microg/mL for 72 h or 144 h, there was a significant reduction in HBeAg and HBsAg expression as well as the HBV DNA copies. CONCLUSION: pGEX-4T-1/HMGN2 vector was success constructed, and the recombinant HMGN2 protein could inhibit HBV expression and replication in vitro remarkably.