Cloning and expression of the nucleoprotein of peste des petits ruminants virus in baculovirus for use in serological diagnosis

Virology. 1995 Apr 20;208(2):776-8. doi: 10.1006/viro.1995.1210.

Abstract

Peste des petits ruminants (PPR) is a viral disease of goats and sheep characterized by erosive stomatitis, enteritis, and pneumonia. The virus is a member of the family Paramyxoviridae and the genus Morbillivirus. The disease has high morbidity and mortality rates and has a substantial economic impact in developing countries. We have cloned and sequenced the cDNA of the nucleocapsid (N) gene of the Nigeria 75/1 strain of PPR virus (PPRV). A comparison of its nucleotide and deduced amino acid sequence with those of the N gene of the tissue culture-attenuated strain of PPRV was performed. A divergence of 8.9 and 5.0% was found at the nucleotide and amino acid level, respectively. A recombinant baculovirus that expresses the N protein in insect cells and larvae (Spodoptera frugiperda) was generated. The recombinant protein, characterized by Western blot analysis, was shown to have a molecular weight of 58 kDa and was recognized by anti-PPRV antibodies. The recombinant protein was used successfully as a coating antigen in an enzyme-linked immunosorbent assay for the serological diagnosis of PPRV.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Viral / blood
  • Baculoviridae / genetics*
  • Base Sequence
  • Capsid / biosynthesis
  • Capsid / chemistry
  • Capsid / genetics*
  • Cell Line
  • Cloning, Molecular
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Gene Expression
  • Genes, Viral / genetics
  • Genetic Variation / genetics
  • Genetic Vectors / genetics
  • Molecular Sequence Data
  • Molecular Weight
  • Morbillivirus Infections / diagnosis*
  • Morbillivirus Infections / veterinary
  • Nigeria
  • Peste-des-petits-ruminants virus / genetics*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Sequence Analysis, DNA
  • Spodoptera
  • Viral Core Proteins / biosynthesis
  • Viral Core Proteins / chemistry
  • Viral Core Proteins / genetics*
  • Viral Structural Proteins / genetics

Substances

  • Antibodies, Viral
  • Recombinant Proteins
  • Viral Core Proteins
  • Viral Structural Proteins