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Article: A truncated hepatitis E virus ORF2 protein expressed in tobacco plastids is immunogenic in mice

TitleA truncated hepatitis E virus ORF2 protein expressed in tobacco plastids is immunogenic in mice
Authors
Issue Date2006
PublisherBaishideng Publishing Group. The Journal's web site is located at http://www.wjgnet.com/1007-9327/index.htm
Citation
World Journal Of Gastroenterology, 2006, v. 12 n. 2, p. 306-312 How to Cite?
AbstractAim: To cost-effectively express the 23-ku pE2, the most promising subunit vaccine encoded by the E2 fragment comprising of the 3'-portion of hepatitis E virus (HEV) open reading frame 2 (ORF2) in plastids of tobacco (Nicotiana tabacum cv. SR1), to investigate the transgene expression and pE2 accumulation in plastids, and to evaluate the antigenic effect of the plastid-derived pE2 in mice. Methods: Plastid-t argeting vector pRB94-E2 containing the E2 fragment driven by rice psbA promoter was constructed. Upon delivery into tobacco plastids, this construct could initiate homologous recombination in psaB-trnfM and trnG-psbC fragments in plastid genome, and result in transgene inserted between the two fragments. The pRB94-E2 was delivered with a biolistic particle bombardment method, and the plastid-transformed plants were obtained following the regeneration of the bombarded leaf tissues on a spectinomycin-supplemented medium. Transplastomic status of the regenerated plants was confirmed by PCR and Southern blot analysis, transgene expression was investigated by Northern blot analysis, and accumulation of pE2 was measured by ELISA. Furthermore, protein extracts were used to immunize mice, and the presence of the pE2-reactive antibodies in serum samples of the immunized mice was studied by ELISA. Results: Transplastomic lines confirmed by PCR and Southern blot analysis could actively transcribe the E2 mRNA. The pE2 polypeptide was accumulated to a level as high as 13.27 μg/g fresh leaves. The pE2 could stimulate the immunized mice to generate pE2-specific antibodies. Conclusion: HEV-E2 fragment can be inserted into the plastid genome and the recombinant pE2 antigen derived is antigenic in mice. Hence, plastids may be a novel source for cost-effective production of HEV vaccines. © 2006 The WJG Press. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/178931
ISSN
2015 Impact Factor: 2.787
2015 SCImago Journal Rankings: 1.076
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorZhou, YXen_US
dc.contributor.authorLee, MYTen_US
dc.contributor.authorNg, JMHen_US
dc.contributor.authorChye, MLen_US
dc.contributor.authorYip, WKen_US
dc.contributor.authorZee, SYen_US
dc.contributor.authorLam, Een_US
dc.date.accessioned2012-12-19T09:50:49Z-
dc.date.available2012-12-19T09:50:49Z-
dc.date.issued2006en_US
dc.identifier.citationWorld Journal Of Gastroenterology, 2006, v. 12 n. 2, p. 306-312en_US
dc.identifier.issn1007-9327en_US
dc.identifier.urihttp://hdl.handle.net/10722/178931-
dc.description.abstractAim: To cost-effectively express the 23-ku pE2, the most promising subunit vaccine encoded by the E2 fragment comprising of the 3'-portion of hepatitis E virus (HEV) open reading frame 2 (ORF2) in plastids of tobacco (Nicotiana tabacum cv. SR1), to investigate the transgene expression and pE2 accumulation in plastids, and to evaluate the antigenic effect of the plastid-derived pE2 in mice. Methods: Plastid-t argeting vector pRB94-E2 containing the E2 fragment driven by rice psbA promoter was constructed. Upon delivery into tobacco plastids, this construct could initiate homologous recombination in psaB-trnfM and trnG-psbC fragments in plastid genome, and result in transgene inserted between the two fragments. The pRB94-E2 was delivered with a biolistic particle bombardment method, and the plastid-transformed plants were obtained following the regeneration of the bombarded leaf tissues on a spectinomycin-supplemented medium. Transplastomic status of the regenerated plants was confirmed by PCR and Southern blot analysis, transgene expression was investigated by Northern blot analysis, and accumulation of pE2 was measured by ELISA. Furthermore, protein extracts were used to immunize mice, and the presence of the pE2-reactive antibodies in serum samples of the immunized mice was studied by ELISA. Results: Transplastomic lines confirmed by PCR and Southern blot analysis could actively transcribe the E2 mRNA. The pE2 polypeptide was accumulated to a level as high as 13.27 μg/g fresh leaves. The pE2 could stimulate the immunized mice to generate pE2-specific antibodies. Conclusion: HEV-E2 fragment can be inserted into the plastid genome and the recombinant pE2 antigen derived is antigenic in mice. Hence, plastids may be a novel source for cost-effective production of HEV vaccines. © 2006 The WJG Press. All rights reserved.en_US
dc.languageengen_US
dc.publisherBaishideng Publishing Group. The Journal's web site is located at http://www.wjgnet.com/1007-9327/index.htmen_US
dc.relation.ispartofWorld Journal of Gastroenterologyen_US
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subject.meshAnimalsen_US
dc.subject.meshBlotting, Northernen_US
dc.subject.meshBlotting, Southernen_US
dc.subject.meshHepatitis E Virus - Immunologyen_US
dc.subject.meshMiceen_US
dc.subject.meshMice, Inbred Balb Cen_US
dc.subject.meshPlastids - Geneticsen_US
dc.subject.meshPolymerase Chain Reactionen_US
dc.subject.meshRecombinant Proteins - Immunologyen_US
dc.subject.meshTobacco - Geneticsen_US
dc.subject.meshVaccines, Synthetic - Immunologyen_US
dc.subject.meshViral Hepatitis Vaccines - Immunologyen_US
dc.subject.meshViral Proteins - Genetics - Immunologyen_US
dc.titleA truncated hepatitis E virus ORF2 protein expressed in tobacco plastids is immunogenic in miceen_US
dc.typeArticleen_US
dc.identifier.emailChye, ML: mlchye@hkucc.hku.hken_US
dc.identifier.emailYip, WK: wkyip@hkucc.hku.hken_US
dc.identifier.authorityChye, ML=rp00687en_US
dc.identifier.authorityYip, WK=rp00833en_US
dc.description.naturepublished_or_final_versionen_US
dc.identifier.doi10.3748/wjg.v12.i2.306-
dc.identifier.pmid16482635-
dc.identifier.pmcidPMC4066044-
dc.identifier.scopuseid_2-s2.0-33644760771en_US
dc.identifier.hkuros113419-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-33644760771&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume12en_US
dc.identifier.issue2en_US
dc.identifier.spage306en_US
dc.identifier.epage312en_US
dc.identifier.isiWOS:000239946500021-
dc.publisher.placeChinaen_US
dc.identifier.scopusauthoridZhou, YX=23136063700en_US
dc.identifier.scopusauthoridLee, MYT=12766751800en_US
dc.identifier.scopusauthoridNg, JMH=12767250100en_US
dc.identifier.scopusauthoridChye, ML=7003905460en_US
dc.identifier.scopusauthoridYip, WK=7102784428en_US
dc.identifier.scopusauthoridZee, SY=7004001733en_US
dc.identifier.scopusauthoridLam, E=7102890014en_US

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