传染性造血器官坏死病病毒糖蛋白杆状病毒表达系统的构建及其抗原性分析

为研究传染性造血器官坏死病病毒（infectious haematopoietic necrosis virus，IHNV）主要结构蛋白糖蛋白（G），本研究采用RT-PCR方法从IHNV中提取RNA并进行反转录，经PCR方法扩增获得1 380 bp的G蛋白基因片段，将其克隆到pFB-LIC-Bse杆状病毒载体中，成功构建了重组质粒pFB-LIC-Bse-G，转化到大肠杆菌DH10Bac感受态细胞中，获得了重组杆粒rBacmid-G。将重组杆粒转染至Sf9昆虫细胞，获得了重组杆状病毒。间接免疫荧光（IFA）和Western blotting分析显示，重组G蛋白可与抗组氨酸单抗（Anti-His）、抗IHNV鼠血清发生特异性反应，表明本研究克隆的IHNV G蛋白在真核表达系统中得到正确表达，且该蛋白具有良好的抗原性。本试验结果为研究G蛋白的功能及开发IHNV新型疫苗奠定了基础。

Construction of Baculovirus Expression System and Antigenicity Analysis of Infectious Hematopoietic Necrosis Virus Glycoprotein

To study the major structural protein glycoprotein (G) of infectious hematopoietic necrosis virus (IHNV), glycoprotein gene (1 380 bp) was amplified by RT-PCR from IHNV. In order to construct a recombinant plasmid pFB-LIC-Bse-G, G gene was cloned into the baculovirus vector pFB-LIC-Bse. Then, the constructed plasmid pFB-LIC-Bse-G was transformed into E.coli DH10Bac. The recombinant bacmids rBacmid-G was got, and then it was transfected to insect Sf9 cells,and the recombinant baculovirus that contained G gene was obtained. Western blotting analysis and indirect immunofluorescence assay (IFA) showed that the recombinant G protein could be recognized by histidine monoclonal antibody (Anti-His) and anti-IHNV antibody of mouse. The results indicated IHNV G protein had been expressed correctly in Sf9 cells. The study laid a foundation for further studying the G protein structure, function and immunological characteristics.