Cloning and Expression of the Nucleoprotein of Peste des Petits Ruminants Virus in Baculovirus for Use in Serological Diagnosis

Tarek M. Ismail, Miles K. Yamanaka, Jeremiah T. Saliki, Alaa El-Kholy, Charles Mebus, Tilahun Yilma

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39 Scopus citations

Abstract

Peste des petits ruminants (PPR) is a viral disease of goats and sheep characterized by erosive stomatitis, enteriris, 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.

Original languageEnglish (US)
Pages (from-to)776-778
Number of pages3
JournalVirology
Volume208
Issue number2
DOIs
StatePublished - Apr 20 1995

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ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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