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Title: Characterization and in vitro expression of non-structural 1 protein of canine parvovirus (CPV-2) in mammalian cell line
Authors: Saxena, Lovleen
Chaturvedi, Uttara
Saxena, Shikha
Kumar, G Ravi
Sahoo, A P
Kumar, Sudesh
Doley, J
Rajmani, R S
Singh, Prafull K
Kumar, Rajiv
Tiwari, Ashok K
Keywords: Eukaryotic expression vector;Canine parvovirus;NS-1 gene;Non-structural protein 1;Polymerase chain reaction
Issue Date: Sep-2011
Publisher: NISCAIR-CSIR, India
Abstract: Parvoviruses are small, 260-Å-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.
Page(s): 654-659
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Appears in Collections:IJEB Vol.49(09) [September 2011]

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