1. Prokaryotic Expression of F Protein from PPRV and Characterization of Its Polyclonal Antibody.
- Author
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Wang, Qiuxia, Dou, Yongxi, Yang, Xiangfang, Meng, Xuelian, Zhai, Junjun, Zhu, Xueliang, Luo, Xuenong, Chen, Lei, and Cai, Xuepeng
- Subjects
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MONOCLONAL antibodies , *PESTE des petits ruminants , *PROKARYOTIC genomes , *RECOMBINANT proteins , *ANTIGENS - Abstract
The goal of this study was to evaluate the specificity of a polyclonal antibody against the F protein from Peste des petits ruminants virus (PPRV). A pET30a/F prokaryotic expression vector was successfully constructed and its recombinant protein was expressed. The result of Western blot analysis showed that the fusion protein pET30a/F possessed good immunoreactivity and the purified recombinant protein was then used as the antigen to raise anti-pET30a/F polyclonal antibody in rabbits. The polyclonal antibody titer against the recombinant F protein was confirmed by indirect ELISA, and the protein's specificity against pET30/F polyclonal antibody was confirmed by both Western blot and indirect immunofluorescence assay in transfected cells. In short, we obtained the high-level expression of recombinant F protein as well as high titers of rabbit polyclonal antibody specificity against F protein in pCAGGS/F transfected cells. This special polyclonal antibody offers a valuable and useful tool for further study of the pathogenesis of PPRV early infection and the structural and functional characterization of PPRV F protein. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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