[Replication and encapsidation of HBV mutants with the truncated C gene].
OBJECTIVE
To evaluate the replication and encapsidation of HBV mutants with the truncated C gene.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Han
Ju-qiang
JQ
Institute of Hepatology, Beijing Military General Hospital, Beijing 100700, China.
Hu
Da-rong
DR
Xiong
Jin-hua
JH
Hu
Xue-ling
XL
Fan
Gong-ren
GR
Li
Juan
J
Liu
Chao-ying
CY
Di
Yi-pin
YP
Wu
Yi-pin
YP
D045744:Cell Line, Tumor; D006512:Hepatitis B Core Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D014162:Transfection; D014779:Virus Replication
D006512:Hepatitis B Core Antigens