猪源H9N2亚型流感病毒HA基因的序列测定及真核表达  

Sequencing and eukaryotic expression of HA gene of H9N2 subtype influenza virus from swine

作　　者：韩庆功[1] 乔传玲[1] 杨焕良[1] 陈艳[1] 辛晓光[1] 丁选亚[1] 陈化兰[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/农业部动物流感重点开放实验室

出　　处：《中国预防兽医学报》2007年第12期916-919,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家科技攻关计划项目(2004BA519A55)

摘　　要：利用RT-PCR扩增猪流感病毒A/Swine/Zhejiang/7/2004(H9N2)(SW/ZhJ7/04)HA基因,测序后将其克隆到真核表达载体pCAGGS上,经酶切分析、PCR鉴定和测序验证,得到重组表达质粒pCAGGS-HA,之后将其转染293T细胞,48h后收集转染细胞样品,进行SDS-PAGE分析,结果显示HA蛋白获得了表达。经Western blot检测,结果表明pCAGGS-HA能够在体外瞬时表达具有免疫学活性的HA蛋白,通过免疫小鼠,对其免疫效果进行了初步研究。HA gene of SW/ZhJ7/04 was amplified by RT-PCR, sequenced, and cloned into recombinant plasmid pCAGGS-HA. The 293T cells monolayer were transfected with plasmid pCAGGS-HA, expression of the HA protein was detected by SDS-PAGE 48 h after transfection and its immunogenic activity was confirmed by Western blot assay. The immunogenicity of the plasmid pCAGGS-HA was evaluated in mice.

关 键 词：猪流感病毒 HA基因 序列测定 真核表达 

分 类 号：S852.659.5[农业科学—基础兽医学]