Identification of a conserved B-cell epitope on the GapC protein of Streptococcus dysgalactiae.
Streptococcus dysgalactiae (S. dysgalactia) GapC is a highly conserved surface dehydrogenase among the streptococcus spp., which is responsible for inducing protective antibody immune responses in animals. However, the B-cell epitope of S. dysgalactia GapC have not been well characterized. In this study, a monoclonal antibody 1F2 (mAb1F2) against S. dysgalactiae GapC was generated by the hybridoma technique and used to screen a phage-displayed 12-mer random peptide library (Ph.D.-12) for mapping the linear B-cell epitope. The mAb1F2 recognized phages displaying peptides with the consensus motif TRINDLT. Amino acid sequence of the motif exactly matched (30)TRINDLT(36) of the S. dysgalactia GapC. Subsequently, site-directed mutagenic analysis further demonstrated that residues R31, I32, N33, D34 and L35 formed the core of (30)TRINDLT(36), and this core motif was the minimal determinant of the B-cell epitope recognized by the mAb1F2. The epitope (30)TRINDLT(36) showed high homology among different streptococcus species. Overall, our findings characterized a conserved B-cell epitope, which will be useful for the further study of epitope-based vaccines.
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China; College of Animal Science and Veterinary Medicine, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Limeng
L
Zhang
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Xue
X
Zhou
College of Animal Science and Veterinary Medicine, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Ziyao
Z
Fan
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Wei
W
Tang
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Liang
L
Chen
College of Animal Science and Veterinary Medicine, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Jian
J
Dai
College of Animal Science and Veterinary Medicine, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Yuhua
Y
Wei
College of Animal Science and Veterinary Medicine, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Jianxin
J
Zhang
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Xuan
X
Yang
College of Animal Science and Veterinary Medicine, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Xijing
X
Yang
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Daolong
D
Liu
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Liquan
L
Yu
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Hua
H
Zhang
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Zhijun
Z
Wu
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Yongzhong
Y
Yu
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China.
Hunan
H
Sun
College of Life Science and Technology, HeiLongJiang BaYi Agricultural University, Daqing 163319, China; College of Animal Science and Veterinary Medicine, HeiLongJiang BaYi Agricultural University, Daqing 163319, China. Electronic address: cuiyudong@yahoo.com.
Yudong
Y
Cui
D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D017124:Conserved Sequence; D004252:DNA Mutational Analysis; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed; D019151:Peptide Library; D017385:Sequence Homology; D013291:Streptococcus / Q000235:genetics / Q000276:immunology*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D018985:Epitopes, B-Lymphocyte; C469817:GapC protein, Streptococcus; D019151:Peptide Library
Epitope; GapC; Homology; Phage display; Streptococcus dysgalactiae