Abstract: The aim of present study was to investigate whether the amino acid replacement of VP60 could affect the expression of VP60 and the capable of self-assembling into VLPs. In order to obtain the chimeric protein of VP60 carrying double OVA T-cell epitopes,the sequences of VP60 302-309aa were replaced by the genes of double OVA T-cell epitopes by SOE. The fused genes were cloned into the donor vector pFastBac^TMHT 1 and the recombinant baculoviruse(rAc-V-302-309) was constructed using Bac-to-Bac baculovirus expression system. The chimeric protein named DC was expressed effectively in insect cells and confirmed by IFA,SDS-PAGE,Western Blot and RTPCR. The chimeric protein was correctly expressed in baculovirus system and could self-assemble into VLPs by Electron microscopy analysis. This manuscript should be very valuable for enhancing the understanding of using RHDV-VLPs as a carrier for foreign genes. Meanwhile,the length of the foreign genes which RHDV-VLPs carried was extended.

Key words: RHDV, VP60, Double OVA T-cell epitopes, VLPs

CLC Number: 

• S858.2912.65