title: Localization of rabies virus glycoprotein into the endoplasmic reticulum produces immunoprotective antigen.
abstract: Rabies virus surface glycoprotein (rabies G-protein) with (G+RS) and without (G-RS) endoplasmic reticulum retrieval signal was expressed and characterized in tobacco plants. Transgenically expressed rabies G-protein was estimated at 0.015-0.38 % of total leaf protein. The relative migration of the rabies G-protein on SDS-PAGE was at the position, as anticipated for the viral coat protein (~66 kDa). Immunolocalization by confocal microscopy established that immunoprotective G+RS expressed in tobacco was primarily confined to ER. G+RS showed binding to Con A lectin and was susceptible to N-glycosidase F activity similar to native rabies G-protein. However, the G-RS transgenically expressed in tobacco leaves was glycosylated differently and was resitant to N-glycosidase F. Immunological studies and Rapid Fluorescent Foci Inhibition Test (RFFIT) showed that G+RS was immunogenic and immunoprotective, whereas G-RS was moderately immunogenic but non-protective against live virus challenge. Hence, plants can express the antigenic component of rabies virus with suitable glycosylation, which is important to give protection against rabies virus infection.
authors.affiliation: National Botanical Research Institute, Rana Pratap Marg, Lucknow 226001, India. yadav_dinesh2002@rediffmail.com
authors.forename: Dinesh K
authors.initials: DK
authors.lastname: Yadav
authors.forename: Shadma
authors.initials: S
authors.lastname: Ashraf
authors.forename: Pradhyumna K
authors.initials: PK
authors.lastname: Singh
authors.forename: Rakesh
authors.initials: R
authors.lastname: Tuli
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D004926:Escherichia coli; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006031:Glycosylation; D051379:Mice; D008807:Mice, Inbred BALB C; D018515:Plant Leaves / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D021381:Protein Transport; D011818:Rabies / Q000517:prevention & control; D011819:Rabies Vaccines / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011820:Rabies virus / Q000276:immunology; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism; D014611:Vaccination; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins; D021382:Protein Sorting Signals; D011819:Rabies Vaccines; D014759:Viral Envelope Proteins; C064951:glycoprotein G, Rabies virus