title: Expression and antigenicity characterization for truncated capsid protein of porcine circovirus type 2.
abstract: Three pairs of specific primers were designed to amplify F2-1, F2-2, and XF2-2 truncated capsid protein genes of porcine circovirus type 2 (PCV-2). Amplified sequences were subcloned to pET-32a(+) vectors and expressed in Rosetta (DE3) Escherichia coli by induction of isopropy-β-D-thiogalactoside (IPTG). All of the fusion proteins had positive reactions to PCV-2 antiserum and His-XF2-2 showed the best reactivity. Proteins were used to immunize BALB/c mice to produce monoclonal antibodies (mAbs), and 7 mAbs were selected. Capsid protein N-terminal parts 55 to 96 amino acid (aa), 97 to 141 aa, and 143 to 211 aa were confirmed as binding regions of the 7 mAbs. Reactivity between His-XF2-2 and the 7 mAbs was detected, FmAb-8 showed the best reactivity. The dominant B-cell epitope was located at 97 to 141 aa. The PEPSCAN indicated that the P122-136 peptide contained the dominant B-cell epitope.
authors: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, China.
authors: Zhongzi
authors: Z
authors: Lou
authors: Xuerui
authors: X
authors: Li
authors: Zhiyong
authors: Z
authors: Li
authors: Xiangping
authors: X
authors: Yin
authors: Baoyu
authors: B
authors: Li
authors: Xi
authors: X
authors: Lan
authors: Bin
authors: B
authors: Yang
authors: Yun
authors: Y
authors: Zhang
authors: Jixing
authors: J
authors: Liu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001666:Binding Sites, Antibody; D036022:Capsid Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017925:Circovirus / Q000235:genetics / Q000378:metabolism*; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D018985:Epitopes, B-Lymphocyte; D011993:Recombinant Fusion Proteins