title: Characterization and in vitro expression of non-structural 1 protein of canine parvovirus (CPV-2) in mammalian cell line.
abstract: Parvoviruses are small, 260-A-diameter, icosahedral, non-enveloped, single-stranded DNA viruses with a genome of approximately 5 kb. Non structural protein, (NS-1) is especially relevant, being both essential for virus replication and the main factor responsible for virus pathogenicity and cytotoxicity. This protein has also been reported to possess the property of killing of transformed cells. The present study was carried out to clone, characterize and express the NS-1 gene of canine parvovirus. NS-1 complete CDS 2020bp was amplified, cloned into eukaryotic expression vector pcDNA 3.1(+), sequenced and characterized by in vitro expression analysis. Functional activity of recombinant construct, pcDNA.cpv.NS-1, was evaluated by RT-PCR and flow cytometry for the expression of NS-1 specific mRNA and NS-1 protein, respectively, in transfected HeLa cells. This recombinant plasmid may serve as an important tool to evaluate the apoptotic potential of NS-1 protein of canine parvovirus in cultured HeLa cells.
authors.affiliation: Molecular Biology Laboratory, Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar 243 122, India.
authors.forename: Lovleen
authors.initials: L
authors.lastname: Saxena
authors.forename: Uttara
authors.initials: U
authors.lastname: Chaturvedi
authors.forename: Shikha
authors.initials: S
authors.lastname: Saxena
authors.forename: G Ravi
authors.initials: GR
authors.lastname: Kumar
authors.forename: A P
authors.initials: AP
authors.lastname: Sahoo
authors.forename: Sudesh
authors.initials: S
authors.lastname: Kumar
authors.forename: J
authors.initials: J
authors.lastname: Doley
authors.forename: R S
authors.initials: RS
authors.lastname: Rajmani
authors.forename: Prafull K
authors.initials: PK
authors.lastname: Singh
authors.forename: Rajiv
authors.initials: R
authors.lastname: Kumar
authors.forename: Ashok K
authors.initials: AK
authors.lastname: Tiwari
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D017993:Parvovirus, Canine / Q000235:genetics*; D010957:Plasmids; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology
chemical_list: D004274:DNA, Recombinant; D004279:DNA, Viral; D017361:Viral Nonstructural Proteins