Replication and encapsidation of HBV mutants with the truncated C gene / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
; (6): 39-42, 2004.
Article
in Zh
| WPRIM
| ID: wpr-281811
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the replication and encapsidation of HBV mutants with the truncated C gene.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>
Full text:
1
Index:
WPRIM
Main subject:
Physiology
/
Plasmids
/
Virus Replication
/
Transfection
/
Hepatitis B virus
/
Cell Line, Tumor
/
Genetics
/
Hepatitis B Core Antigens
/
Mutation
Limits:
Humans
Language:
Zh
Journal:
Chinese Journal of Experimental and Clinical Virology
Year:
2004
Type:
Article