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Chinese Journal of Experimental and Clinical Virology ; (6): 39-42, 2004.
Artículo en Chino | WPRIM | ID: wpr-281811

RESUMEN

<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>


Asunto(s)
Humanos , Línea Celular Tumoral , Antígenos del Núcleo de la Hepatitis B , Genética , Virus de la Hepatitis B , Genética , Fisiología , Mutación , Plásmidos , Genética , Transfección , Replicación Viral
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