表达猪H1N1亚型流感病毒三聚体HA的重组慢病毒包装  

Packaging of recombinant lentivirus expressing trimeric HA of swine H1N1 influenza virus

作　　者：李鑫[1] 宋战昀[2] 朱利塞[1] 樊娇[1] 王广美[1] 丛彦龙[1] 丁壮[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]吉林省出入境检验检疫局,吉林长春130062

出　　处：《中国兽医学报》2012年第12期1828-1831,1845,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金青年基金项目(31101825)

摘　　要：以RT—PCR扩增猪H1N1流感病毒HA基因，用GS-linker使HA与GCN4pⅡ连接。构建慢病毒蛋白表达系统。通过观察荧光、病毒转导效率、SDS—PAGE、Western blot验证重组慢病毒包装和HA—GCN4pⅡ融合蛋白表达情况。该系统能使外源基因得到持久和稳定的表达。结果，包装获得能够融合表达HA-GCN4pⅡ的重组慢病毒LV—HA-GCN4pⅡ，病毒滴度为1．1×10^6TU／mL；LV—HA—GCN4pⅡ能有效转导293T细胞。本试验成功构建了表达HA—GCN4PⅡ融合蛋白的慢病毒表达载体，获得的重组慢病毒不仅为后续动物试验测定其诱导的体液免疫和细胞免疫水平奠定了基础，同时也为研制猪H1N1亚型流感疫苗做出了有益尝试。To package a recombinant lentivirus which can express HA-GCN4p Ⅱ fusion protein. The HA gene derived from swine H1N1 influenza virus was amplified by RT-PCR and linked to GCN4p Ⅱ by a GS-linker. The expression of HA GCN4p Ⅱand packaging of recombinant lentivirus were identified by the results of green fluorescence,transduction efficiency, SDS-PAGE, and Western blot. A recombinant lentivirus LV-HA-GCN4p Ⅱ was achieved with a titer of 1.1 × 10^6 TU/mL,which can be transduced in 293T ceils efficiently. The achievement of a recombinant lentivirus is not only beneficial to further prepare a swine H1N1 influenza vaccine,but also helpful to pave the way for constructing a pseudotyped lentiviral system stably expressing HA-GCN4p Ⅱ.

关 键 词：猪流感 慢病毒载体 HA GCN4pⅡ 

分 类 号：S852.65[农业科学—基础兽医学]