title: Development and in vitro characterization of a bivalent DNA containing HN and F genes of velogenic Newcastle disease virus.
abstract: Newcastle disease (ND) is highly contagious, economically important viral disease affecting most of avian species worldwide. Newcastle disease virus (NDV) has single stranded negative sense RNA genome which encodes for six structural and two non-structural proteins. Envelope glycoproteins i.e. hemagglutinin-neuraminidase (HN) and the fusion (F), elicit protective immune response. In this study, HN and F genes of velogenic (virulent) strain were amplified and cloned at multiple cloning sites A and B, respectively into pIRES bicistronic vector for use as bivalent DNA vaccine against ND. The recombinant plasmid was characterized for its orientation by restriction enzyme digestion and PCR. Expression of HN and F genes was assessed in transfected Vero cells at RNA level using RT-PCR in total RNA as well as protein level using IFAT, IPT and western blot using NDV specific antiserum. All these experiments confirmed that HN and F genes cloned in recombinant pIRES.nd.hn.f are functionally active. The recombinant construct is being evaluated as DNA vaccine against ND.
authors.affiliation: Molecular Biology Laboratory, Division of Animal Biotechnology Indian Veterinary Research Institute, Izatnagar 243 122, India.
authors.forename: Uttara
authors.initials: U
authors.lastname: Chaturvedi
authors.forename: Shahina
authors.initials: S
authors.lastname: Kalim
authors.forename: G
authors.initials: G
authors.lastname: Desai
authors.forename: Barkha
authors.initials: B
authors.lastname: Ratta
authors.forename: Rajiv
authors.initials: R
authors.lastname: Kumar
authors.forename: P V
authors.initials: PV
authors.lastname: Ravindra
authors.forename: Sudesh
authors.initials: S
authors.lastname: Kumar
authors.forename: B B
authors.initials: BB
authors.lastname: Dash
authors.forename: Sangeeta
authors.initials: S
authors.lastname: Tiwari
authors.forename: A P
authors.initials: AP
authors.lastname: Sahoo
authors.forename: Ashok K
authors.initials: AK
authors.lastname: Tiwari
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D015707:HN Protein / Q000235:genetics*; D009522:Newcastle disease virus / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014162:Transfection; D019444:Vaccines, DNA / Q000235:genetics* / Q000276:immunology; D014709:Vero Cells; D014760:Viral Fusion Proteins / Q000235:genetics* / Q000276:immunology
chemical_list: D004279:DNA, Viral; D015707:HN Protein; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D014760:Viral Fusion Proteins