Expression of EV71-VP1 Protein and Preparation of Its Polyclonal Antibody

摘要

The objective of this study is to utilize the protein VP1 of Enterovirus 71 from clinical specimens of children with hand-foot-and-mouth disease (HFMD) to prepare the polyclonal antibody (pAb), prevented the viral infection. RNA got out from the diachorema of the sufferer was amplified by RT-PCR and nested PCR. The viral capsid protein (VP1) cDNA was inserted into the vector PET32a(+), an expression vector PET32a-VPl was constructed. The VP1 protein was expressed in E Coli. and induced by IPTG . The polyclonal antibody (pAb) against the fusion protein was prepared. ELISA and Western blot methods were used to identify the titer and specificity of the pAb. The VP1 protein and the polyclonal antibody were prepared successfully. The successful expression of the capsid protein VP1 of EV71 and the preparation of the polyclonal antibody can afford experiment foundation for the deep research of the biological function.