Construction of eukarotic expression vector of HBV gene with nt 1862 mutation and its expression in Cos7 cells

Objective To study the biological significance of HBV gene mutation at nucleotide (nt) 1862. Methods The EB virus eukarotic expression vector for HBV pre-C/C gene was constructed using molecular biological method, HBV pre-C/C gene mutation at nt 1862 was induced by way of site-specific mutation technique, and identified by PCR-RFLP and sequencing analysis. The resulted recombinant plasmid containing the HBV variant was subsequently transfected into Cos7 cell line mediated by lipofectin, to observe the expression of HBeAg. The cells transfected with the recombinant plasmid containing wild HBV Pre-C/C gene fragment served as control. Results HBeAg expression was detected in the cells transfected with wild recombinant plasmid but not in those with HBV variant transfection. Conclusion The success in the construction of eukarotic expression vector for HBV pre-C/C gene mutation at nt 1862 may pave the way for further studying a series of biological changes of HBV resulted from the mutation addressed in this study.