title: Expression of rabies virus glycoprotein gene into eukaryotic system and determination of potential T-cell epitopes.
abstract: The present study was undertaken to clone, express rabies virus glycoprotein (RVG) and to identify potential T-cell epitopes on it. RVG gene (1590 bp) was amplified using gene specific primers. The amplified product was cloned into pTZ57R/T cloning vector by TA cloning. RVG gene was subcloned into pcDNA3.1 (+) expression vector. In this study, cloning and expression of rabies virus glycoprotein gene was done under CMV promoter and an expression construct (pcDNA.RVG) was prepared and clones were confirmed by restriction digestion, colony PCR and nucleotide sequencing. The expression construct was further characterized by western blotting and indirect fluorescent antibody test (IFAT). In silico analysis of this protein was done to find out potential antigenic sites so that it can be further evaluated for its potential as candidate for epitope vaccine against rabies.
authors.affiliation: Department of Molecular Biology and Genetic Engineering, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar 263145, India. nehatomar.biotech@gmail.com
authors.forename: Neha R
authors.initials: NR
authors.lastname: Tomar
authors.forename: Rajesh
authors.initials: R
authors.lastname: Chandra
authors.forename: Rajiv
authors.initials: R
authors.lastname: Kumar
authors.forename: A K
authors.initials: AK
authors.lastname: Tiwari
authors.forename: Anil
authors.initials: A
authors.lastname: Kumar
mesh_terms: D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D006224:Cricetinae; D004587:Electrophoresis, Agar Gel; D018984:Epitopes, T-Lymphocyte / Q000276:immunology*; D005822:Genetic Vectors / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics*; D051379:Mice; D010957:Plasmids; D011819:Rabies Vaccines / Q000235:genetics / Q000276:immunology; D011820:Rabies virus / Q000235:genetics* / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D018984:Epitopes, T-Lymphocyte; D006023:Glycoproteins; D011819:Rabies Vaccines; D014764:Viral Proteins