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Replication and encapsidation of HBV mutants with the truncated C gene / 中华实验和临床病毒学杂志
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>
Subject(s)
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
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