中文摘要：

在兔出血症病毒（RHDV）主要结构蛋白VP60的N端插入双串联卵清蛋白（OVA）CD8^＋T细胞表位（OVA第257-264位氨基酸）,研究外源片段对VP60蛋白表达及病毒样颗粒（VLPs）装配的影响,分析VP60作为载体的可行性和对外源基因长度的耐受性。通过分子生物学方法将双串联OVA CD8^＋T细胞表位（257-264aa）插入至RHDV衣壳蛋白VP60基因序列的N端,得到重组VP60基因。利用杆状病毒表达系统表达嵌合蛋白,并命名为DN。经RT-PCR、间接免疫荧光、SDS-PAGE、Western blot方法鉴定嵌合蛋白DN的表达,利用电镜观察嵌合蛋白病毒样颗粒的形成。结果表明嵌合VP60蛋白在杆状病毒表达系统中得到高效表达,且可形成与RHDV VLPs类似的病毒样颗粒。本研究为VP60作为载体系统,有效提呈外源片段的研究提供了重要依据,同时也为VLPs疫苗的研制奠定了基础。

英文摘要：

The aim of present study was to investigate whether the amino acid insertion of VP60 could affect the expression of VP60 and the ability of self-assembling into virus-like particles （VLPs）. In order to obtain the chimeric protein of VP60 carrying double ovalbumin （OVA） T-cell epitopes, the fragment of double OVA T-cell epitopes was inserted into the N-terminus of VP60. The recombinant baculovirus （ rAc -V-DN） was constructed using Bac-to-Bac baculovirus expression system. The chimeric protein named DN was expressed effectively in insect cells and confirmed by RT-PCR, IFA, SDS-PAGE and Western blot. The chimeric protein was correctly expressed and could self-assemble into VLPs by Electron microscopy analysis. This study is very valuable for enhancing the understanding of using RHDV-VLPs as a carrier for foreign genes. Meanwhile, it would be useful for the development of VLPs vaccine in future.