H5亚型AIV HA1基因与鸡IL-18基因共表达载体的构建及其表达  

Construction and expression of recombinant vector coexpressing HA1 gene from subtype H5 of avian influenza virus and chicken interleukin-18 gene

作　　者：孔娜[1] 田夫林 兰邹然 杨林 冯涛 梁成珠[3] 赵宏坤[1] 

机构地区：[1]山东农业大学动物科技学院,山东泰安271018 [2]山东省畜牧局,山东济南250022 [3]山东出入境检验检疫局,山东青岛266001

出　　处：《中国兽医学报》2009年第8期986-990,1007,共6页Chinese Journal of Veterinary Science

基　　金：山东省科技攻关重大资助项目"山东省畜禽重大疫病防治技术研究"(SDSP2005041005);国家"十一五"科技支撑计划重大资助项目"食品安全的综合示范"(2006BAK02A21)

摘　　要：参考GenBank上发表的H5亚型禽流感(AIV)血凝素(HA)基因序列及鸡白细胞介素18成熟蛋白(mChIL-18)的cDNA基因序列,分别设计了HA1和mChIL-18基因的2对引物。然后分别以pMD18-T-HA质粒和pGEX-mChIL-18质粒为模板,扩增出了H5亚型AIV的HA1基因和mChIL-18的cDNA片段。再以杆状病毒pFast-BacTMdual为载体,将H5亚型AIV HA中的HA1基因和mChIL-18基因分别插入到双表达载体pFastBacTMdu-al的p10启动子(PP10)和多角体蛋白启动子(PPH)的下游,构建携带HA1基因和mChIL-18基因的重组pFastBacTMdual-HA1-mChIL-18真核表达质粒。最后将构建的真核表达质粒转座入大肠杆菌DH10Bac,并利用昆虫细胞进行转染。这个重组载体的构建为考察mChIL-18作为免疫增强剂的作用及下一步AIV疫苗的研究奠定了坚实的基础。The hemagglutinin（HA） gene of avian influenza virus（AIV） subtype H5 and mature chicken interleu- kin-18（mChIL-18） gene were amplified by designing two pairs of primers according to the sequence published on GenBank. Recombinant haculovirus coexpressing HA1 gene from AIV subtype H5 plus chicken interleukin-18 gene were constructed by inserting cDNA copy of HA gene and cDNA copy of mChIL-18 gene into the baculovirus expression plasmid pFast BacTMDual, furthermore cDNAs inserted into the downstreams of P10 promoter and PH promoter,respectively. Finally,the constructing eukaryotic expression plasmid pFastBacTM dual-HAl-mChIL-18 was transposited into Bacterium coli DH10Bac, which is a base of next step insect cell＇s transfection. The construction of this recombinant plasmid could be beneficial to investigate the effect of mChIL-18＇s immunologic enhancement and study a new AIV recombinant vaccine.

关 键 词：禽流感病毒 HA1基因 鸡白细胞介素18成熟蛋白基因 真核表达 转染 

分 类 号：S852.65[农业科学—基础兽医学] R535[农业科学—兽医学]