猪流感复合多表位核酸疫苗的构建及其表达研究  

Construction and Expression of Porcine Influenza Virus Multi-epitope DNA Vaccine

作　　者：贾雷立[1] 金宁一[2] 金扩世[2] 郑敏[1] 金洪涛[1] 连海[1] 王瑞琳[1] 李昌[1] 鲁会军[2] 马鸣潇[1] 金明兰[1] 

机构地区：[1]吉林大学畜牧兽医学院预防兽医学系,长春130062 [2]军事医学科学院军事兽医研究所病毒室,长春130062

出　　处：《中国生物制品学杂志》2006年第4期333-335,共3页Chinese Journal of Biologicals

基　　金：国家科技攻关项目(2004BA519A12);长春市振兴老工业基地科技攻关项目(04-02GG217).

摘　　要：目的构建猪流感病毒复合多表位核酸疫苗,并研究其表达产物的抗原性。方法以H3、H9亚型流感的HA抗原表位及流感病毒的其它主要抗原(NP、NA、M)表位基因为基础,再附以Kozak序列和适当的酶切位点,设计并合成一段长765bp的复合多表位基因(Epi),其中各表位基本独立,将其克隆入真核表达载体pIRES1neo,构建重组质粒pIRE-Epi。将多表位抗原基因Epi插入到融合表达基因H57中,构建重组质粒pIRE-H57-Epi。将构建的重组质粒在Hela细胞中表达,并用免疫荧光方法初步检测所表达蛋白的抗原性。结果所构建的融合表达载体pIRE-Epi和pIRE-H57-Epi,在Hela细胞中表达出流感病毒多表位抗原蛋白,并具有抗原性。结论已成功构建猪流感病毒多表位基因,有可能成为较理想的猪流感候选疫苗。Objective To construct porcine influenza virus multi-epitope DNA vaccine and study the antigenicity of expressed product. Methods Design and synthesize a DNA fragment containing the HA antigenic epitopes of influenza virus subtype H3 and H9 as well as the epitopes of other major antigens（ NP, NA and M ） of influenza virus, Kozak sequence and proper restriction sites ,in which the epitopes were separated by spacer sequence（ -P-P-S-）, and clone into eukaryotic expression vector pIRES1 neo to construct recombinant plasmid pIRE-Epi. Clone multi-epitope antigen gene Epi into plasmid pUCM-T-H57 to construct a transfer vector pUCM-T-H57- Epi,then clone H57-Epi into pIRES1neo to construct expression vector pIRE-H57-Epi. Transformed the constructed recombinant plamids to Hela cells,and determine the antigenicity of expressed protein by IFA. Results Both fusion expression vectors pIRE-Epi and pIRE-H57-Epi were successfitlly constructed,and porcine influenza virus multi-epitope antigen with antigenicity was expressed in Hela cells. Conclusion Porcine influenza virus multi-epitope DNA was successfully constructed,which might be an ideal candidate for porcine influenza vaccine.

关 键 词：流感 表位 核酸疫苗 

分 类 号：R511.7[医药卫生—内科学] R392.11[医药卫生—临床医学]