【作者】 贾永清； 乔传玲； 张建林； 于康震； 刘宝全；

【Author】 Jia Yongqing, Qiao Chuanling, Zhang Jianlin ,Yu Kangzhen, Liu Baoquan( Animal Influenza Research Center, Harbin Veterinary Research Institute of CAAS, Harbin 150001)( State Key Laboratory of Veterinary Biotecnology of Harbin VeterinaryResearch Institute ,CAAS, Harbin 150001) ( College of Veterinary Medicine Southeast Agricultural University, Harbin 150030)

【机构】 中国农业科学院哈尔滨兽医研究所动物流感研究中心； 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； 东北农业大学动物医学院；

【摘要】 将AIV广东分离株A/Goose/Guangdong/3/96(H5N1)HA基因正向克隆于pSY538的EcoR Ⅰ位点,将LacZ报告基因插入Sma Ⅰ位点,然后切取含外源基因和报告基因的Not Ⅰ片段克隆于pSY681的Not Ⅰ位点,构建AIV HA基因重组禽痘病毒转移载体,经酶切分析和PCR鉴定后,利用脂质体转染试剂盒在鸡胚成纤维细胞上与F-017禽痘病毒弱毒疫苗株进行共转染。经重组病毒筛选与纯化,获得遗传性状稳定的AIV HA基因重组禽痘病毒。更多还原

【Abstract】 In order to develop a recombinant fowlpox virus (FPV) expressing avian influenza virus (AIV) haemagglutinin (HA), the HA cDNA of H5 subtype AIV A/Goose/Guangdong/3/ 96(H5N1)and LacZ reporter gene were cloned into pSY538 plasmid and then subcloned into Not I site of pSY681 plasmid. After construction of transfer vector, it was transfected on CEF cells with FPV F-017. Following 6 cycle screenings, the pure HA-fowlpox virus recombinants were obtained, and identified by restriction enzyme analysis and PCR. The results showed that the recombinant fowlpox virus was stable on genetic properties.更多还原