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- W2364246906 abstract "Objective To construct expression vector containing HBV rtN236T and rtA181V/T mutations and explore their replication capacity and susceptibility to adefovir after transient transfection into HepG2 cells.Methods The plasmid containing the entire HBV(1.2 copies)genome was used to construct adefovir-resistant expression plasmids via PCR-based site-directed mutagenesis in vitro.The constructed plasmids were validated by DNA sequencing and served as positive controls for mutant-specific PCR.After transient transfection into HepG2 cells,HBsAg and HBeAg in supernatants were measured by ELISA,HBV DNA levels in supernatants and Intracellular HBV replicative intermediates were measured by Florescence Real-time PCR and Southern blotting,respectively To compare the replication capacity and susceptibility to adefovir.Results Plasmids containing adefovir resistance mutations of rtA181T/V or rtN236T were constructed and expressed in HepG2 cells.These plasmids can be specifically positive control for mutant- specific PCR.In the transfection experiment,mutation with rtA181T/V or rtN236T could independently confer resistance to adefovir in vitro,ICSO of which were 2.8 to 4.7 folds more than wild type.Also, rtA181T/V or rtN236T mutants showed decreases of replication capacities,which were 94.2%,89.0% and 77.7% of wild type HBV,respectively.Conclusion HBV with either rtA181T/V or rtN236T can confer resistance to adefovir resulting in decreased replication capacity." @default.
- W2364246906 created "2016-06-24" @default.
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- W2364246906 date "2007-01-01" @default.
- W2364246906 modified "2023-09-22" @default.
- W2364246906 title "The construction of HBV strains with rtN236T and rtA181V/T mutations associated with resistance to adefovir and its viral characteristics" @default.
- W2364246906 hasPublicationYear "2007" @default.
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