重组杆状病毒表达H9亚型禽流感HA基因的研究  

Expression of H9 Subtype Avian Influenza HA Gene in Baculovirus System

作　　者：石霖[1,2] 熊永忠[2] 李晓楠[1] 赵晓彤[1] 顾贵波[1] 谷志大[1] 邓炜娜[3] 薛树山[1] 吴运谱[1] 王宏燕[1] 魏澍[1] 张雅为[1] 王竹[1] 曹东[1] 

机构地区：[1]辽宁省动物疫病预防控制中心,辽宁沈阳110164 [2]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150000 [3]第四军医大学西京医院,陕西西安710000

出　　处：《河南农业科学》2014年第1期120-122,126,共4页Journal of Henan Agricultural Sciences

基　　金：辽宁省博士启动基金项目(20121148)

摘　　要：为研究通过体外表达的方式获得具有生物学活性的禽流感HA蛋白的方法,根据GenBank公布的禽流感H9亚型的基因序列设计引物,扩增出HA基因全长片段,将扩增片段克隆到pFastBacHTa载体上,构建重组杆粒rBacmid-HA,在脂质体介导下转染Sf9细胞制备重组杆状病毒,表达HA蛋白,并通过血凝抑制试验、Westernt-blot、间接免疫荧光试验研究其生物学活性。结果显示,在Sf9细胞中成功表达了HA蛋白,分子量约为65kD,且表达蛋白具有良好的生物学活性。This study intends to express HA protein of avian influenza in vitro ,and lay the foun-dation for the serological diagnosis method which distinguish avian influenza subtypes .The pri-mers were designed according to sequences of subtype H 9 avian influenza virus in GenBank ,am-plified fragment of HA gene ,and amplified fragments were cloned into pFastBacHTa vector .The recombinant plasmid was transformed into DH10 competent cells to produce recombinant bacmid rBacmid-HA ,recombinant baculovirus was obtained through the liposome mediated transfection of Sf9 cells ,and the target protein was expressed in the Sf9 cells ,the molecular weight of about 65 kD .By hemagglutination test ,Westernt-blot ,indirect immuno fluorescence test showed that the protein was successfully expressed ,and had a good biological activity .

关 键 词：禽流感 H9亚型 HA基因 杆状病毒 表达 

分 类 号：S852.657[农业科学—基础兽医学]