Prokaryotic expression of NS1' protein resulted from ribosomal frameshifting in Japanese encephalitis virus and its immunoreactivity

The NS1' gene fragment resulted from ribosomal frameshifting in Japanese encephalitis virus(JEV) was amplified by RT-PCR and inserted into a prokaryotic expression vector pET-32a.The recombinant plasmid pET-32a-NS1' was transformed into Escherichia coli BL21 and induced to express by IPTG.The fusion protein with a molecular weight of 25ku was visible on the SDS-PAGE gel and purified with His-Bind Purification Kit.The Western blotting analysis showed that the NS1' protein could react with the polyclonal antibody from the mice immunized with wild strain of JEV,while the NS1' protein did not react with that from mice immunized with JEV vaccine.These results indicated that the NS1' protein had the potential to distinguish between wild strain infection and vaccine immunization in clinical practice.