H3N8亚型EIV HA基因在昆虫细胞中的表达及生物学特性分析  

Expression and biological characterization of HA protein of H3N8 subtype equine influenza virus in insect cells

作　　者：刘春国[1] 王伟[1] 刘飞[2] 刘彦云[1] 王丹[3] 吕让[1] 刘明[1] 相文华[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]中国农业科学院上海兽医研究所,上海200241 [3]黑龙江省医院,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2013年第11期934-936,共3页Chinese Journal of Preventive Veterinary Medicine

摘　　要：为表达H3N8亚型马流感病毒(EIV)HA蛋白,本研究将A/equine/xinjiang/3/2007(H3N8)的HA基因克隆至杆状病毒转移载体pFastBac1中,构建重组转移质粒pFB-XJ3-HA,并将其转化至DH10BAC感受态细胞中,与杆状病毒骨架质粒Bacmid进行重组,重组杆粒rBac-XJ3-HA转染Sf9昆虫细胞,构建重组杆状病毒rBv-XJ3-HA,并通过细胞免疫组化和western blot进行鉴定。结果表明rBv-XJ3-HA构建正确,表达的HA蛋白与抗EIV的血清反应良好;血凝试验结果表明表达的HA蛋白具有良好的血凝活性。本研究为研制EIV HA蛋白亚单位疫苗提供了实验依据。To express the hemagglutinin （HA） protein of H3N8 subtype equine influenza virus （EIV）, the HA gene of E1Vs A/equine/xinjiang/3/2007 （H3N8） was sub-cloned into transfer plasmid pFastBacl and then the recombinant shuttle plasmid pFB-XJ3-HA was transformed into DH10BA competent cells to generate a recombinant baculovirus bacmid of rBac-XJ3-HA. The rBac-XJ3-HA was transfected into Sf9 insect cells for packing a recombinant baculovirus of rBv-XJ3-HA which was confirmed by PCR, immunohistochemistry, western blot, and hemagglutination test. The results showed that HA protein of E1V was expressed in insect cells and have the immunoreactivity and hemagglutinin activity, which provide the basis for the development of subunit vaccine of H3N8 subtype equine influenza.

关 键 词：马流感病毒 H3N8亚型 HA基因 杆状病毒表达 血凝活性 

分 类 号：S852.65[农业科学—基础兽医学]