兔出血症病毒衣壳蛋白在昆虫细胞中的表达及对家兔的免疫保护效果  

Expression of the Capsid Protein of Rabbit Haemorrhagic Disease Virus in Insect Cells and Its Protective Efficacy to Rabbits

作　　者：王芳[1] 胡波[1] 任雪枫[2] 范志宇[1] 杨龙圣[1] 徐为中[1] 张则斌[1] 何孔旺[1] 

机构地区：[1]江苏省农业科学院兽医研究所,南京210014 [2]江苏省动物疫病预防控制中心,南京210036

出　　处：《畜牧兽医学报》2008年第10期1382-1387,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：江苏省科技成果转化专项资金(BA2004032-020);江苏省农业科学院院基金(025056110639)

摘　　要：用RT—PCR方法扩增兔出血症病毒（RHDV）衣壳蛋白VP60基因，通过克隆、转化得到重组穿梭载体Bacmid—VP60，用此质粒转染Sf9昆虫细胞，得到重组病毒rAcV—Bac—VP60，经RT—PCR、IFA、SDS-PAGE、Western blotting、HA和HI鉴定，结果显示重组VP60蛋白在Sf9昆虫细胞中得到表达。在不加任何佐剂的情况下，用表达的蛋白免疫3月龄非RHD免疫兔，结果显示，免疫后21天，兔体内可产生抗RHDV抗体，抗体血凝抑制效价为2^6～2^7，该抗体可以抵抗血凝效价为2^10致死剂量RHDV强毒的攻击，本研究为兔病毒性出血症基因工程疫苗的研制奠定了基础。The objective of this study is to develop genetic engineering vaccine to rabbit haemorrhagic disease. RT-PCR -was used to amplify the capsid protein VP60 gene of rabbit haemorrhagic disease virus （RHDV）. After being cloned and transformed, the recombinant shuttle vector, Bacmid-VP60, was constructed and transfected into Sf9 insect cells, then a recombinant baculovirus, named rAcV-Bac-VP60 was obtained. The identification by IFA, SDS-PAGE, Western blotting, HA and HI showed that the recombinant protein of VP60 was expressed in Sf9 insect cells. The expressed protein was applied to immunize three-month-old rabbits that without immunizing with RHD vaccine. The results showed that the anti-RHDV antibodies were induced and developed after 21 days of immunization, the anti-RHDV titers in serum ranged from 2^6 to 2^7. When challenged with 2^10 HA units of infectious RHDV, the control rabbits died, while the immunized animals survived. The results indicated that the recombinant proteins expressed by recombinant baculovirus could be ideal candidate for RHDV vaccines.

关 键 词：兔出血症病毒 衣壳蛋白 杆状病毒表达系统 重组杆状病毒 表达 免疫 

分 类 号：S858.291[农业科学—临床兽医学] S432.41[农业科学—兽医学]