H9N2亚型禽流感病毒病毒样颗粒的构建

Construction of Virus-Like Particles of H9N2 Avian Influenza Virus

关键词:  禽流感病毒  H9N2亚型  病毒样颗粒  Bac-to-Bac杆状病毒表达系统

Key words:  Avian influenza virus  subtype H9N2  virus-like particles  Bac-to-Bac baculovirus expression system

摘要:

为构建H9N2亚型禽流感病毒病毒样颗粒，本研究以血凝素（Hemagglutinin，HA）、神经氨酸酶（Neurami-nidase，NA）及基质蛋白（Matrix protein，M1）3种蛋白为对象，通过Bac-to-Bac杆状病毒表达系统构建了2种重组杆状病毒，即rBV-HA-M1和rBV-NA，用IFA和Western-blot鉴定重组蛋白的免疫活性，再将rBV-HA-M1和rBV-NA以最佳感染复数比例共感染昆虫Sf9细胞，经浓缩后获得毒样颗粒，同时进行血凝滴度测定和透射电镜观察。透射电镜观察结果显示，表达的HA-M1和NA重组蛋白可以在Sf9细胞中自组装形成病毒样颗粒，浓缩的病毒样颗粒能够凝集1%的鸡红细胞，血凝效价为1:64。试验结果为进一步研究H9N2亚型禽流感病毒病毒样颗粒疫苗奠定了基础。

Abstract:

In order to construct virus-like particles of H9N2 avian influenza virus, this study used the proteins of hemagglutinin (HA), neuraminidase (NA) and matrix protein (M1) to construct two recombinant baculoviruses rBV-HA-M1 and rBV-NA, through the Bac-to-Bac baculovirus expression system. The immune activity of the recombinant proteins were identified by IFA and Western-blot, then the Sf9 insect cells were co-infected with rBV-HA-M1 and rBV-NA at the optimal infection complex ratio. The co-infected Sf9 cells were concentrated to obtain the virus-like particles. The virus-like particles were measured for hemagglutination titers and transmission electron microscopy. The results of transmission electron microscopy showed that the expressed HA-M1 and NA recombinant proteins could self-assemble to form virus-like particles in Sf9 cells. The virus-like particles were able to agglutinate 1% of chicken red blood cells, and the titer was 1:64. The results of this study lay the foundation for further research on the virus-like particle vaccine of H9N2 virus.