中文摘要：

利用卵清蛋白（OVA）第257～264位氨基酸替换兔出血症病毒的主要结构蛋白质VP60不同位置处相应数量的氨基酸，研究了氨基酸的替换对VP60蛋白表达及病毒样颗粒装配的影响。通过重叠延伸剪接术（SOE）法，OVAT-细胞表位（257～264位氨基酸）分别取代VP603个区域（502～510位氨基酸、411～417位氨基酸、302—309位氨基酸），得到3种重组嵌合VP60基因。利用杆状病毒表达系统，得到3种重组穿梭载体Bacmid，分别命名为Bacmid—V-1、Bacmid-V-2、Bacmid—V-3。将重组阳性质粒转染Sf9昆虫细胞，得到3种重组病毒rBac-v-1、rBac-v-2、rBac-v-3，3组重组病毒感染Sf9细胞后表达的3种蛋白质分别命名为z1、z2和z3。经liT—PCR、IFA、SDS—PAGE、Westernblot等方法鉴定后，利用电镜观察此3种重组蛋白质病毒样颗粒的形成情况。3种重组嵌合VP60蛋白在Sf9昆虫细胞中得到表达，3种重组嵌合蛋白质均可以形成与VP60蛋白类似的病毒样颗粒。

英文摘要：

The amino acids 257-264 on OVA （ovalbumin） was used as a substitute for corresponding number of amino acids on different locations of VP60, the major structural protein of rabbit hemorrhagic disease virus, to study the effects of the substitution on the expression of VP60 and VLPs （virus-like particles） assembling. By splicing by overlap ex- tension （ SOE ） method, three areas on VP60, amino acids 502- 510, amino acids 411 - 417, and amino acids 302- 309 were replaced by OVA T-cell epitope （OVA 257-264） to get three recombinant chimeric VP60 genes. Via the baculovirus expression system, three recombinant shuttle vector bacmids, named Bacmid-V-1, Bacmid-V-2 and Bacmid-V-3 were obtained. The positive recombinant plasmids were transfected into Sf 9 insect cells to produce three kinds of recombinant viruses, rBac-v-1, rBac-v-2, rBac-v- 3, and three kinds of proteins expressed in the recombinant virus infected Sf 9 cells were named Z1, Z2, and Z3. After identification by RT-PCR, IFA, SDS-PAGEand Western blot, three recombinant proteins were observed under electron microscopy. All the three recombinant chimeric VP60 proteins wer expressed in Sf 9 insect cells, and formed VLPs that similar toVP60.