Cloning and expression of the nucleoprotein of peste des petits ruminants virus in baculovirus for use in serological diagnosis.
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.
Ismail
T M
TM
Department of Veterinary Pathology, Microbiology and Immunology, University of California, Davis, USA.
Yamanaka
M K
MK
Saliki
J T
JT
el-Kholy
A
A
Mebus
C
C
Yilma
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D002213:Capsid / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D002460:Cell Line; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015870:Gene Expression; D005814:Genes, Viral / Q000235:genetics; D014644:Genetic Variation / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D018185:Morbillivirus Infections / Q000175:diagnosis* / Q000662:veterinary; D009549:Nigeria; D018107:Peste-des-petits-ruminants virus / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D017422:Sequence Analysis, DNA; D018411:Spodoptera; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D015678:Viral Structural Proteins / Q000235:genetics
D000914:Antibodies, Viral; D011994:Recombinant Proteins; D014758:Viral Core Proteins; D015678:Viral Structural Proteins