猪圆环病毒2b型Cap蛋白多克隆抗体制备及其生物学特性分析

doi:10.16431/j.cnki.1671-7236.2015.05.002

Abstract

Abstract: In order to build more effective use of porcine circovirus type 2b (PCV2b) Cap protein on the diagnosis and control of diseases like porcine circovirus disease, the ORF2 gene which encoding porcine circovirus type 2b Cap protein was cloned into the prokaryotic expression vector pET-32a (+), constructing a recombinant plasmid pET-32a-ORF2, then the recombinant plasmid was transformed into E.coli BL21 and induced by IPTG to express recombinant protein, the recombinant protein was purified with nickel column.Then polyclonal antibody (PcAb) was prepared by immunization of rabbit with purified protein.ELISA, Western blotting, indirect immunofluorescence assay and the specificity experiments were used to detect the biological characteristics of the polyclonal antibody.The titer of polyclonal antibody was about 1:2¹⁶ detected by ELISA.Western blotting result confirmed that polyclonal antibody could react with PCV2b specially.Indirect immunofluorescence assay showed that polyclonal antibody was able to detect PCV2b in PK-15 cells, this result suggested that the polyclonal antibody had the ability to differential diagnosis of PCV2b.The specificity experiment showed that the polyclonal antibody only reacted with PCV2b, and did not react with PEDV, TGEV, PRRSV, PRoV, PRV and PPV.The polyclonal antibody against PCV2b prepared in this study provided a powerful tool for the study of etiological characteristics and clinic diagnosis of this disease.

Key words: porcine circovirus type 2b; Cap protein; prokaryotic expression; polyclonal antibody; biological characteristics

CLC Number:

S852.65

YUAN Qing, DONG Bo, GAO Rui-ze, LI Peng-chong, REN Yu-dong, REN Xiao-feng, LI Guang-xing. Preparation of Polyclonal Antibody against Porcine Circovirus Type 2b Cap Protein and Analysis of its Biological Characteristics[J]. , 2015, 42(5): 1048-1055.

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Preparation of Polyclonal Antibody against Porcine Circovirus Type 2b Cap Protein and Analysis of its Biological Characteristics

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