1. Cloning and expression of the nucleoprotein of peste des petits ruminants virus in baculovirus for use in serological diagnosis.
- Author
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Ismail TM, Yamanaka MK, Saliki JT, el-Kholy A, Mebus C, and Yilma T
- Subjects
- Amino Acid Sequence, Animals, Antibodies, Viral blood, Base Sequence, Capsid biosynthesis, Capsid chemistry, Cell Line, Cloning, Molecular, Gene Expression, Genes, Viral genetics, Genetic Variation genetics, Genetic Vectors genetics, Molecular Sequence Data, Molecular Weight, Morbillivirus Infections veterinary, Nigeria, Recombinant Proteins biosynthesis, Recombinant Proteins chemistry, Sequence Analysis, DNA, Spodoptera, Viral Core Proteins biosynthesis, Viral Core Proteins chemistry, Viral Structural Proteins genetics, Baculoviridae genetics, Capsid genetics, Enzyme-Linked Immunosorbent Assay methods, Morbillivirus Infections diagnosis, Peste-des-petits-ruminants virus genetics, Viral Core Proteins genetics
- 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.
- Published
- 1995
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