Replication and encapsidation of HBV mutants with the truncated C gene
Han, J.-q.; Hu, D.-r.; Xiong, J.-h.; Hu, X.-l.; Fan, G.-r.; Li, J.; Liu, C.-y.; Di, Y.-p.; Wu, Y.-p.
Zhonghua Shi Yan he Lin Chuang Bing du Xue Za Zhi 18(1): 39-42
2004
ISSN/ISBN: 1003-9279 PMID: 15340524 Document Number: 575259
To evaluate the replication and encapsidation of HBV mutants with the truncated C gene. The HBV mutants with the truncated C gene were constructed by molecular cloning and PCR-based deletion in vitro. The replication and encapsidation of HBV mutants were investigated by Southern blotting, PCR and real-time fluorescence PCR respectively after transfecting the HBV mutants plasmid into HepG2 cells by using liposome. The C-truncated HBV mutant vectors were constructed successfully and confirmed exactly by clone sequencing and enzymes digestion. The C-truncated HBV mutants were replication defective, however, all types of HBV DNA could be detected positive in the cytoplasm and supernatant after co-transfecting the C-truncated HBV mutants plasmid and the helper constructs into HepG2 cells. The C-truncated HBV mutants were proved to produce 3-40 folds more progeny DNA than that of the wild-type HBV by DNA quantitative assay. The C-truncated HBV mutants are replication-deficient and could not replicate and encapsulate in the hepatocytes when transfected solely, however, the progeny HBV-variant viruses are encapsidated more effectively to secrete into supernatant when co-transfected with the helper construct which lacks part of 5 prime-proximal HBV RNA packaging signal Epsilon.