质粒驱动的流感病毒样颗粒的形成。
Plasmid-driven formation of influenza virus-like particles.
作者信息
Neumann G, Watanabe T, Kawaoka Y
机构信息
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
出版信息
J Virol. 2000 Jan;74(1):547-51. doi: 10.1128/jvi.74.1.547-551.2000.
Abstract
We established a plasmid-based system for generating infectious influenza virus-like particles entirely from cloned cDNAs. Human embryonic kidney cells (293T) were transfected with plasmids encoding the influenza A virus structural proteins and with a plasmid encoding an influenza virus-like viral RNA (vRNA) which contained an antisense copy of the cDNA for green fluorescence protein (GFP) flanked by an RNA polymerase I promoter and terminator. Intracellular transcription of the latter construct by RNA polymerase I generated GFP vRNA that was packaged into influenza virus-like particles. This system, which produced more than 10(4) infectious particles per ml of supernatant, would be useful in studies of influenza virus replication and particle formation. It might also benefit efforts in vaccine production and in the development of improved gene therapy vectors.
摘要
我们建立了一种基于质粒的系统，可完全从克隆的cDNA产生传染性流感病毒样颗粒。用人胚胎肾细胞（293T）转染编码甲型流感病毒结构蛋白的质粒以及编码流感病毒样病毒RNA（vRNA）的质粒，该vRNA含有绿色荧光蛋白（GFP）cDNA的反义拷贝，两侧为RNA聚合酶I启动子和终止子。RNA聚合酶I对后一种构建体进行细胞内转录，产生GFP vRNA，其被包装到流感病毒样颗粒中。该系统每毫升上清液可产生超过10^4个感染性颗粒，将有助于流感病毒复制和颗粒形成的研究。它也可能有助于疫苗生产以及改进基因治疗载体的开发。
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