Localization of Antigenic Sites at the Amino-Terminus of Rinderpest Virus N Protein Using Deleted N Mutants and Monoclonal Antibody

J Vet Sci. 2003 Aug;4(2):167-73.

Abstract

The nucleocapsid (N) protein of rinderpest virus (RPV) is highly conserved, immunogenic, and abundantly expressed during infection. Six antigenic sites (sites A, B, C, D, E and F), defined previously by a competitive binding assay using corresponding monoclonal antibodies (Mabs), have been further localized by immunoassays using deleted N mutants. Five different forms of RPV N protein, containing residues aa 1-79, aa 1-149, aa 1-421, aa 414-525 and aa 1-525, were expressed as glutathione S transferase (GST) fusion proteins (designated as GST-N1-79, GST-N1-149, GST-N1-421, GST-N414-525, and GST-N1-525, respectively) in E.coli BL21 cells. In ELISA using deleted N mutants, Mabs recognizing sites A, B, C, D and E reacted with 3 GST fusion proteins (GST-N1-149, GST-N1-421 and GST-N1-525), indicating that they are located at aa 80-149. Mab recognizing site F reacted with 4 GST fusion proteins (GST-N1-79, GST-N1-149, GST-N1-421 and GST-N1-525), indicating that site F is located at aa 1-79. Identification of the amino-terminal antigenic sites of the N protein would provide antigen basis for developing sensitive and specific diagnostic reagents for RPV, although it remains to be further investigated antigenic sites at the carboxyl-terminus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal
  • Base Sequence
  • Chlorocebus aethiops
  • Cloning, Molecular
  • DNA Primers
  • Escherichia coli / genetics
  • Molecular Sequence Data
  • Nucleocapsid Proteins / analysis
  • Nucleocapsid Proteins / chemistry
  • Nucleocapsid Proteins / genetics*
  • Recombinant Proteins / chemistry
  • Rinderpest virus / chemistry
  • Rinderpest virus / genetics*
  • Rinderpest virus / isolation & purification
  • Sequence Alignment
  • Sequence Deletion
  • Sequence Homology, Amino Acid
  • Vero Cells
  • Viral Proteins / analysis
  • Viral Proteins / chemistry
  • Viral Proteins / genetics*

Substances

  • Antibodies, Monoclonal
  • DNA Primers
  • Nucleocapsid Proteins
  • Recombinant Proteins
  • Viral Proteins
  • nucleocapsid protein, Rinderpest virus