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Article: High level virion production and surface antigen expression with 1.5 copies of hepatitis B viral genome
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TitleHigh level virion production and surface antigen expression with 1.5 copies of hepatitis B viral genome
 
AuthorsDong, Q2
Liu, Z2
Chen, Y2
Chan, CY2
Lin, MC1
Kung, Hf2
Chan, HLY2
Sung, JJY2
He, ML2
 
KeywordsBasal core promoter
HBeAg
HBsAg
Hepatitis B virus
Pregenomic RNA
 
Issue Date2009
 
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jviromet
 
CitationJournal Of Virological Methods, 2009, v. 159 n. 2, p. 135-140 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.jviromet.2009.03.012
 
AbstractThe present study aimed to construct a 1.5X hepatitis B virus (HBV) replication system in vitro that could generate high level of HBV viruses. This system would help compare the replication capacity among the virus strains associated with high and low risk of hepatocellular carcinoma (HCC). Four strains of HBV were isolated from two HCC patients and two HBV carriers. After molecular cloning, four corresponding constructs named as HBV-1.5Xs were generated. Each of them has one and a half copies of HBV 3.2 kb genome, a 5′-end redundant sequence of 1.1 kb to nt715 and a 3′-end redundant sequence of 500 bp to nt2325 that situated after the poly (A) sequence. The HepG2 cells were transfected with the HBV-1.5Xs, and the levels of HBsAg, HBeAg and viral DNA were then detected in both the supernatant and the cells. After 24 h and 48 h of transfection, a high OD value of HBsAg of 3.5 was observed in the supernatant and also in some of the diluted cell lysate samples. The HBeAg level was relatively low in all strain samples of HBV. The log 10 values of viral loads were also determined with the cell lysate having a higher value (10-11 per ml) than that of the supernatant (6-7 per ml). The results showed that the novel HBV-1.5X system was capable to generate high level of HBV in a consistent manner. However, no significant difference was found among the replication capacities among these strains in vitro. The HBV-1.5X system may be a useful platform that assists the establishment of stable cell lines and transgenic mice for the investigation of viral pathogenesis, particularly for the various strains of HBV. © 2009 Elsevier B.V. All rights reserved.
 
ISSN0166-0934
2012 Impact Factor: 1.9
2012 SCImago Journal Rankings: 0.740
 
DOIhttp://dx.doi.org/10.1016/j.jviromet.2009.03.012
 
ISI Accession Number IDWOS:000267454000001
Funding AgencyGrant Number
Research Grant Council (RGC), Hong KongCUHK4428/06M
CUHK7334/03M
Funding Information:

This study was supported by grants from the Research Grant Council (RGC), Hong Kong (Nos. CUHK4428/06M and CUHK7334/03M).

 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorDong, Q
 
dc.contributor.authorLiu, Z
 
dc.contributor.authorChen, Y
 
dc.contributor.authorChan, CY
 
dc.contributor.authorLin, MC
 
dc.contributor.authorKung, Hf
 
dc.contributor.authorChan, HLY
 
dc.contributor.authorSung, JJY
 
dc.contributor.authorHe, ML
 
dc.date.accessioned2010-09-06T06:14:44Z
 
dc.date.available2010-09-06T06:14:44Z
 
dc.date.issued2009
 
dc.description.abstractThe present study aimed to construct a 1.5X hepatitis B virus (HBV) replication system in vitro that could generate high level of HBV viruses. This system would help compare the replication capacity among the virus strains associated with high and low risk of hepatocellular carcinoma (HCC). Four strains of HBV were isolated from two HCC patients and two HBV carriers. After molecular cloning, four corresponding constructs named as HBV-1.5Xs were generated. Each of them has one and a half copies of HBV 3.2 kb genome, a 5′-end redundant sequence of 1.1 kb to nt715 and a 3′-end redundant sequence of 500 bp to nt2325 that situated after the poly (A) sequence. The HepG2 cells were transfected with the HBV-1.5Xs, and the levels of HBsAg, HBeAg and viral DNA were then detected in both the supernatant and the cells. After 24 h and 48 h of transfection, a high OD value of HBsAg of 3.5 was observed in the supernatant and also in some of the diluted cell lysate samples. The HBeAg level was relatively low in all strain samples of HBV. The log 10 values of viral loads were also determined with the cell lysate having a higher value (10-11 per ml) than that of the supernatant (6-7 per ml). The results showed that the novel HBV-1.5X system was capable to generate high level of HBV in a consistent manner. However, no significant difference was found among the replication capacities among these strains in vitro. The HBV-1.5X system may be a useful platform that assists the establishment of stable cell lines and transgenic mice for the investigation of viral pathogenesis, particularly for the various strains of HBV. © 2009 Elsevier B.V. All rights reserved.
 
dc.description.natureLink_to_subscribed_fulltext
 
dc.identifier.citationJournal Of Virological Methods, 2009, v. 159 n. 2, p. 135-140 [How to Cite?]
DOI: http://dx.doi.org/10.1016/j.jviromet.2009.03.012
 
dc.identifier.doihttp://dx.doi.org/10.1016/j.jviromet.2009.03.012
 
dc.identifier.epage140
 
dc.identifier.hkuros156069
 
dc.identifier.isiWOS:000267454000001
Funding AgencyGrant Number
Research Grant Council (RGC), Hong KongCUHK4428/06M
CUHK7334/03M
Funding Information:

This study was supported by grants from the Research Grant Council (RGC), Hong Kong (Nos. CUHK4428/06M and CUHK7334/03M).

 
dc.identifier.issn0166-0934
2012 Impact Factor: 1.9
2012 SCImago Journal Rankings: 0.740
 
dc.identifier.issue2
 
dc.identifier.openurl
 
dc.identifier.pmid19490966
 
dc.identifier.scopuseid_2-s2.0-67349205097
 
dc.identifier.spage135
 
dc.identifier.urihttp://hdl.handle.net/10722/69554
 
dc.identifier.volume159
 
dc.languageeng
 
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jviromet
 
dc.publisher.placeNetherlands
 
dc.relation.ispartofJournal of Virological Methods
 
dc.relation.referencesReferences in Scopus
 
dc.rightsJournal of Virological Methods. Copyright © Elsevier BV.
 
dc.subjectBasal core promoter
 
dc.subjectHBeAg
 
dc.subjectHBsAg
 
dc.subjectHepatitis B virus
 
dc.subjectPregenomic RNA
 
dc.titleHigh level virion production and surface antigen expression with 1.5 copies of hepatitis B viral genome
 
dc.typeArticle
 
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Author Affiliations
  1. The University of Hong Kong
  2. Chinese University of Hong Kong