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Article: Use of GFP to investigate expression of plant-derived vaccines.

TitleUse of GFP to investigate expression of plant-derived vaccines.
Authors
Issue Date2009
Citation
Methods In Molecular Biology (Clifton, N.J.), 2009, v. 515, p. 275-285 How to Cite?
AbstractPlants are low-cost bioreactors for the production of various biopharmaceuticals including oral vaccines. Plant-derived oral vaccines are potentially useful in combating viral infections involving mucosal immunity. Transgenic plants have been generated to successfully produce mucosal vaccines against cholera, hepatitis B, foot-and-mouth disease, and Norwalk virus. As a first step toward the generation of oral vaccines against the severe acute respiratory syndrome coronavirus (SARS-CoV), we have expressed a recombinant S1 protein of the SARS-CoV in transformed tobacco. Since plant transformation and regeneration of stable transformants require considerable time, we initially used a green fluorescent protein (GFP) to tag the antigen in transient expression. GFP was fused to the carboxy-terminus of S1 for expression of S1-GFP to show expression of recombinant S1 by agroinfiltration of tobacco leaves. The GFP tag enables a relatively quick confirmation of antigen expression in plant cells by fluorescent microscopy. Such analysis using GFP that precedes stable plant transformation will enable the rapid screening of multiple constructs to attain optimal recombinant protein expression. Furthermore, this approach determines the subcellular localization of the recombinant protein in plant cells, providing information on optimal subcellular targeting for production in plant bioreactors.
Persistent Identifierhttp://hdl.handle.net/10722/179132
ISSN
2015 SCImago Journal Rankings: 0.549
References

 

DC FieldValueLanguage
dc.contributor.authorLi, HYen_US
dc.contributor.authorChye, MLen_US
dc.date.accessioned2012-12-19T09:52:14Z-
dc.date.available2012-12-19T09:52:14Z-
dc.date.issued2009en_US
dc.identifier.citationMethods In Molecular Biology (Clifton, N.J.), 2009, v. 515, p. 275-285en_US
dc.identifier.issn1064-3745en_US
dc.identifier.urihttp://hdl.handle.net/10722/179132-
dc.description.abstractPlants are low-cost bioreactors for the production of various biopharmaceuticals including oral vaccines. Plant-derived oral vaccines are potentially useful in combating viral infections involving mucosal immunity. Transgenic plants have been generated to successfully produce mucosal vaccines against cholera, hepatitis B, foot-and-mouth disease, and Norwalk virus. As a first step toward the generation of oral vaccines against the severe acute respiratory syndrome coronavirus (SARS-CoV), we have expressed a recombinant S1 protein of the SARS-CoV in transformed tobacco. Since plant transformation and regeneration of stable transformants require considerable time, we initially used a green fluorescent protein (GFP) to tag the antigen in transient expression. GFP was fused to the carboxy-terminus of S1 for expression of S1-GFP to show expression of recombinant S1 by agroinfiltration of tobacco leaves. The GFP tag enables a relatively quick confirmation of antigen expression in plant cells by fluorescent microscopy. Such analysis using GFP that precedes stable plant transformation will enable the rapid screening of multiple constructs to attain optimal recombinant protein expression. Furthermore, this approach determines the subcellular localization of the recombinant protein in plant cells, providing information on optimal subcellular targeting for production in plant bioreactors.en_US
dc.languageengen_US
dc.relation.ispartofMethods in molecular biology (Clifton, N.J.)en_US
dc.subject.meshCapsid Proteins - Genetics - Immunology - Metabolismen_US
dc.subject.meshGene Expressionen_US
dc.subject.meshGenes, Reporter - Geneticsen_US
dc.subject.meshGenetic Vectors - Geneticsen_US
dc.subject.meshGreen Fluorescent Proteins - Analysis - Genetics - Metabolismen_US
dc.subject.meshRecombinant Fusion Proteins - Genetics - Metabolismen_US
dc.subject.meshSars Virus - Genetics - Immunology - Metabolismen_US
dc.subject.meshTobacco - Genetics - Metabolismen_US
dc.subject.meshVaccines - Genetics - Immunology - Metabolismen_US
dc.titleUse of GFP to investigate expression of plant-derived vaccines.en_US
dc.typeArticleen_US
dc.identifier.emailChye, ML: mlchye@hkucc.hku.hken_US
dc.identifier.authorityChye, ML=rp00687en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1007/978-1-59745-559-6_19en_US
dc.identifier.pmid19378126-
dc.identifier.scopuseid_2-s2.0-65649123087en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-84867759072&selection=ref&src=s&origin=recordpage-
dc.identifier.volume515en_US
dc.identifier.spage275en_US
dc.identifier.epage285en_US
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridLi, HY=22953303900en_US
dc.identifier.scopusauthoridChye, ML=7003905460en_US

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