Characterization and in vitro expression of non-structural 1 protein of canine parvovirus (CPV-2) in mammalian cell line

Indian J Exp Biol. 2011 Sep;49(9):654-9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cloning, Molecular
  • DNA, Recombinant / genetics
  • DNA, Viral / genetics
  • Flow Cytometry
  • HeLa Cells
  • Humans
  • Molecular Sequence Data
  • Parvovirus, Canine / genetics*
  • Plasmids
  • Reverse Transcriptase Polymerase Chain Reaction
  • Viral Nonstructural Proteins / biosynthesis*
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / physiology

Substances

  • DNA, Recombinant
  • DNA, Viral
  • Viral Nonstructural Proteins