The expression of EV71 VP1 gene and preparation of its antiserum

EV71 is a main virus of hand-foot-moth disease,while VP1 has main antigenic determinant of EV71.In this study,we aimed to prepare VP1 antiserum and detect its titer by cloning and expressing recombinant VP1 gene.Firstly,the prokaryotic expression vector pET32a(+)-VP1 was constructed by using VP1 fragment from PCR product,and the recombinant protein expressed.Then the purified recombinant protein was used to immunize New Zealand rabbits for preparing VP1 antiserum.At the same time,a eukaryotic expression vector of VP1 was constructed with the aim of detecting the antiserum specificity.ELISA revealed that antibody titer of the prepared antiserum was 1∶64 000;Western blot test of eukaryotic expression vector pcDNA3.1(+)-VP1 protein showed that the antiserum could specifically bind with VP1.Preparation of immunogenic VP1 protein and high titer antiserum has laid a foundation for further study on the methods of diagnosis and resistance vaccine.