Expression and antigenicity characterization for truncated capsid protein of porcine circovirus type 2

Lou, Zhongzi; Li, Xuerui; Li, Zhiyong; Yin, Xiangping; Li, Baoyu; Lan, Xi; Yang, Bin; Zhang, Yun; Liu, Jixing

Expression and antigenicity characterization for truncated capsid protein of porcine circovirus type 2

2011

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.

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