表达禽流感病毒血凝素基因的重组禽痘病毒的构建  

Construction of a Recombinant Fowlpox Virus Expressing Hemagglutinin Gene of Avian Influenza Virus

作　　者：陈化兰[1] 马文军[1] 于康震[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,哈尔滨150001

出　　处：《中国农业科学》2000年第5期86-90,共5页Scientia Agricultura Sinica

摘　　要：为构建表达禽流感病毒 ( AIV)血凝素 ( HA)基因的重组禽痘病毒 ,将禽痘病毒启动子LP2 EP2驱动的 H7亚型 AIV HA基因 c DNA和大肠杆菌 Lac Z基因插入禽痘病毒疫苗株 F- 0 1 7的复制非必需区。经蓝斑筛选后 ,对重组病毒又进行了 3次蚀斑克隆。以不同代次重组禽痘病毒核酸为模板 ,利用 H7亚型 AIV HA基因特异的引物进行聚合酶链式反应 ,均扩增出 1条特异的1 .7kb DNA条带。收集不同代次的重组禽痘病毒细胞培养物进行免疫斑点试验 ,均检测到 HA蛋白 ,表明 H7AIV HA在重组禽痘病毒中获得了成功表达。该重组禽痘病毒的构建为In order to construct a recombinant fowlpox virus expressing gene of avian influenza virus (AIV) hemagglutinin (HA), the HA cDNA of H7 subtype AIV A/Afri. Star./Eng Q/983/79(H7N1)and E.coli Lac Z gene, which were regulated by the fowlpox virus derived promoter LP2 EP2, were inserted into the fragment of F 017 fowlpox virus, which had been confirmed as non essential region for the replication of fowlpox, in the order of LP2EP2 LacZ LP2EP2 HA. Following the first cycle screening of the recombinant based on the expression of the LacZ gene, the HA fowlpox recombinants were selected and verified based on the expression of the AIV HA in further generations. A 1.7 kb DNA was amplified by PCR with the AIV HA specific primers from the recombinant virus DNA.The AIV HA was revealed in the recombinant virus suspension by dot enzyme linked immunosorbent assay (Dot ELISA). The results show that the recombinant fowlpox virus is functional in expression of the H7 AIV HA.

关 键 词：禽流感病毒 血凝素基因 重组禽痘病毒 

分 类 号：S855[农业科学—临床兽医学]