The replication, encapsulation and expression of the recombinant HBV vector with the truncated C gene / 中华传染病杂志

ABSTRACT

Objective To evaluate the replication, encapsulation and expression of the recombinant HBV vector with the truncated C gene. Methods C gene truncated HBV vectors were constructed by technologies of molecular clone and PCR based site-directed mutagenesis in vitro. The expression of GFP was observed with the fluorescence microscope after transfection of the recombinant HBV vector plasmids into HepG2 cells by using the liposome method. The capability of HBV vector replication, encapsulation and progeny virus production were examined with semi nested-PCR, native agarose gel electrophoresis blot, quantitative southern blot analysis and the routine PCR assays. Results The HBV vectors with truncated C gene were constructed successfully. The gene was expressed effectively after transfection. The C gene truncated HBV vectors could be replicated and encapsulated in hepatocytes with the helper virus. The encapsulated efficiency were as 4～8 folders as the non C gene truncated HBV vectors by quantitative analysis. In addition, the mature HBV particles carrying the interesting gene of GFP could secrete out from hepatocytes by the help of adjunctive vector. Conclusions The truncation of C gene could improve the encapsulation efficiency of HBV vectors with no effect on the replication and expression of the intact HBV vectors.