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Article: High level virion production and surface antigen expression with 1.5 copies of hepatitis B viral genome

TitleHigh level virion production and surface antigen expression with 1.5 copies of hepatitis B viral genome
Authors
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
Citation
Journal Of Virological Methods, 2009, v. 159 n. 2, p. 135-140 How to Cite?
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.
Persistent Identifierhttp://hdl.handle.net/10722/69554
ISSN
2014 Impact Factor: 1.781
2014 SCImago Journal Rankings: 0.784
ISI Accession Number ID
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).

References

 

DC FieldValueLanguage
dc.contributor.authorDong, Qen_HK
dc.contributor.authorLiu, Zen_HK
dc.contributor.authorChen, Yen_HK
dc.contributor.authorChan, CYen_HK
dc.contributor.authorLin, MCen_HK
dc.contributor.authorKung, Hfen_HK
dc.contributor.authorChan, HLYen_HK
dc.contributor.authorSung, JJYen_HK
dc.contributor.authorHe, MLen_HK
dc.date.accessioned2010-09-06T06:14:44Z-
dc.date.available2010-09-06T06:14:44Z-
dc.date.issued2009en_HK
dc.identifier.citationJournal Of Virological Methods, 2009, v. 159 n. 2, p. 135-140en_HK
dc.identifier.issn0166-0934en_HK
dc.identifier.urihttp://hdl.handle.net/10722/69554-
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.en_HK
dc.languageengen_HK
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/jvirometen_HK
dc.relation.ispartofJournal of Virological Methodsen_HK
dc.rightsJournal of Virological Methods. Copyright © Elsevier BV.en_HK
dc.subjectBasal core promoteren_HK
dc.subjectHBeAgen_HK
dc.subjectHBsAgen_HK
dc.subjectHepatitis B virusen_HK
dc.subjectPregenomic RNAen_HK
dc.titleHigh level virion production and surface antigen expression with 1.5 copies of hepatitis B viral genomeen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0166-0934&volume=159&issue=2&spage=135&epage=140&date=2009&atitle=High+Level+Virion+Production+and+Surface+Antigen+Expression+with+1.5+Copies+of+Hepatitis+B+Viral+Genomeen_HK
dc.identifier.emailLin, MC:mcllin@hkucc.hku.hken_HK
dc.identifier.authorityLin, MC=rp00746en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.jviromet.2009.03.012en_HK
dc.identifier.pmid19490966en_HK
dc.identifier.scopuseid_2-s2.0-67349205097en_HK
dc.identifier.hkuros156069en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-67349205097&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume159en_HK
dc.identifier.issue2en_HK
dc.identifier.spage135en_HK
dc.identifier.epage140en_HK
dc.identifier.isiWOS:000267454000001-
dc.publisher.placeNetherlandsen_HK
dc.identifier.scopusauthoridDong, Q=8437495200en_HK
dc.identifier.scopusauthoridLiu, Z=36066691200en_HK
dc.identifier.scopusauthoridChen, Y=24075600300en_HK
dc.identifier.scopusauthoridChan, CY=22033276600en_HK
dc.identifier.scopusauthoridLin, MC=7404816359en_HK
dc.identifier.scopusauthoridKung, Hf=7402514190en_HK
dc.identifier.scopusauthoridChan, HLY=25722700100en_HK
dc.identifier.scopusauthoridSung, JJY=35405352400en_HK
dc.identifier.scopusauthoridHe, ML=35080389700en_HK

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