Localization of Antigenic Sites at the Amino-terminus of Rinderpest Virus N Protein Using Deleted N Mutants and Monoclonal Antibody.
- Author:
Kang Seuk CHOI
1
;
Jin Ju NAH
;
Young Joon KO
;
Shien Young KANG
;
Yi Seok JOO
Author Information
1. National Veterinary Research and Quarantine service, Ministry of Agriculture and Forestry, 480 Anyang, Gyounggi 430-824, Korea. choiks@nvrqs.go.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- MeSH:
Amino Acid Sequence;
Animals;
Antibodies, Monoclonal;
Base Sequence;
Cercopithecus aethiops;
Cloning, Molecular;
DNA Primers;
Escherichia coli/genetics;
Molecular Sequence Data;
Nucleocapsid Proteins/analysis/chemistry/*genetics;
Recombinant Proteins/chemistry;
Rinderpest virus/chemistry/*genetics/isolation & purification;
Sequence Alignment;
Sequence Deletion;
Sequence Homology, Amino Acid;
Vero Cells;
Viral Proteins/analysis/chemistry/*genetics
- From:Journal of Veterinary Science
2003;4(2):167-173
- CountryRepublic of Korea
- Language:English
-
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.
