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
王粲 1张民秀 1谢芝勋 1李孟 1罗思思 1李丹 1阮志华 1谢丽基 1谢志勤1
作者信息
- 1. 广西壮族自治区兽医研究所,广西兽医生物技术重点实验室,农业农村部中国(广西)-东盟跨境动物疫病防控重点实验室,南宁 530001
- 折叠
摘要
为构建H9N2 亚型禽流感病毒病毒样颗粒,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,经浓缩后获得病毒样颗粒,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.
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引用本文复制引用
基金项目
广西重点研发计划(桂科AB21076004)
广西八桂学者项目(2019A50)
广西重大专项项目(桂科AA23062050)
广西科技先锋队项目(桂农科盟202409-01)
出版年
2024
国家科技期刊平台
NSTL
万方数据