Cloning and Expression of the Nucleocapsid Gene of Virulent Kabete O Strain of Rinderpest Virus in Baculovirus

Use in Differential Diagnosis between Vaccinated and Infected Animals

Tarek Ismall, Shabbir Ahmad, Marian D'Souza-Ault, Mansour Bassiri, Jeremiah Saliki, Charles Mebus, Tilahun Yilma

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Rinderpest (RP) is a viral disease of ruminants with >95% morbidity and mortality. We have cloned the cDNA of the nucleocapsid (N) gone of the virulent Kabete O strain of rinderpest virus (RPVK) and compared its nucleotide and deduced amino acid sequences with those of the N genes of the lapinized strain of rinderpest virus (RPVL), measles virus (MV), and canine distemper virus (CDV). The gene has an open reading frame of 1575 nucleotides and encodes a protein of 525 amino acids with a molecular weight of 59 kDa. The nucleotide sequence of the coding region of the N gone of RPVK is 88.6, 68.9, and 63.2% homologous with N genes of RPVL, MV, and CDV, respectively. We have developed a recombinant baculovirus that expresses the N protein (rRVN) of RPVK in insect cells (Sf9) and larvae (Spodoptera frugiperda). rRVN was used as a coating antigen in an ELISA to distinguish vaccinated animals from those infected with RPV and was also used successfully in the diagnosis of two other morbilliviruses, MV and peste des petits ruminants (PPRV). Crude lysate of a single infected larva (0.2-0.3 g) was sufficient to coat 150 ELISA plates for serological diagnosis of 7200 serum samples in duplicate.

Original languageEnglish (US)
Pages (from-to)138-147
Number of pages10
JournalVirology
Volume198
Issue number1
DOIs
StatePublished - Jan 1994

Fingerprint

Rinderpest virus
Nucleocapsid
Measles virus
Baculoviridae
Canine Distemper Virus
Organism Cloning
Differential Diagnosis
Gene Expression
Larva
Peste-des-Petits-Ruminants
Morbillivirus
Nucleotides
Rinderpest
Enzyme-Linked Immunosorbent Assay
Genes
Sf9 Cells
Spodoptera
Ruminants
Virus Diseases
Open Reading Frames

ASJC Scopus subject areas

  • Infectious Diseases
  • Virology

Cite this

Cloning and Expression of the Nucleocapsid Gene of Virulent Kabete O Strain of Rinderpest Virus in Baculovirus : Use in Differential Diagnosis between Vaccinated and Infected Animals. / Ismall, Tarek; Ahmad, Shabbir; D'Souza-Ault, Marian; Bassiri, Mansour; Saliki, Jeremiah; Mebus, Charles; Yilma, Tilahun.

In: Virology, Vol. 198, No. 1, 01.1994, p. 138-147.

Research output: Contribution to journalArticle

Ismall, Tarek ; Ahmad, Shabbir ; D'Souza-Ault, Marian ; Bassiri, Mansour ; Saliki, Jeremiah ; Mebus, Charles ; Yilma, Tilahun. / Cloning and Expression of the Nucleocapsid Gene of Virulent Kabete O Strain of Rinderpest Virus in Baculovirus : Use in Differential Diagnosis between Vaccinated and Infected Animals. In: Virology. 1994 ; Vol. 198, No. 1. pp. 138-147.
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abstract = "Rinderpest (RP) is a viral disease of ruminants with >95{\%} morbidity and mortality. We have cloned the cDNA of the nucleocapsid (N) gone of the virulent Kabete O strain of rinderpest virus (RPVK) and compared its nucleotide and deduced amino acid sequences with those of the N genes of the lapinized strain of rinderpest virus (RPVL), measles virus (MV), and canine distemper virus (CDV). The gene has an open reading frame of 1575 nucleotides and encodes a protein of 525 amino acids with a molecular weight of 59 kDa. The nucleotide sequence of the coding region of the N gone of RPVK is 88.6, 68.9, and 63.2{\%} homologous with N genes of RPVL, MV, and CDV, respectively. We have developed a recombinant baculovirus that expresses the N protein (rRVN) of RPVK in insect cells (Sf9) and larvae (Spodoptera frugiperda). rRVN was used as a coating antigen in an ELISA to distinguish vaccinated animals from those infected with RPV and was also used successfully in the diagnosis of two other morbilliviruses, MV and peste des petits ruminants (PPRV). Crude lysate of a single infected larva (0.2-0.3 g) was sufficient to coat 150 ELISA plates for serological diagnosis of 7200 serum samples in duplicate.",
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