Goto Simon, Ishida Kotaro, Suzuki Ryosuke, Morita Eiji
Department of Biochemistry and Molecular Biology, Faculty of Agriculture and Life Science, Hirosaki University, 3 Bunkyo-cho, Hirosaki-shi, Aomori, Japan.
Department of Virology II, National Institute of Infectious Diseases, 1-23-1, Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
Bio Protoc. 2020 May 5;10(9):e3606. doi: 10.21769/BioProtoc.3606.
Cells infected with flavivirus release various forms of infectious and non-infectious particles as products and by-products. Comprehensive profiling of the released particles by density gradient centrifugation is informative for understanding viral particle assembly. However, it is difficult to detect low-abundance minor particles in such analyses. We developed a method for viral particle analysis that integrates a high-sensitivity split luciferase system and density gradient centrifugation. This protocol enables high-resolution profiling of particles produced by cells expressing Japanese encephalitis virus factors.
感染黄病毒的细胞会释放各种形式的感染性和非感染性颗粒,作为产物和副产物。通过密度梯度离心对释放的颗粒进行全面分析,有助于理解病毒颗粒的组装。然而,在这类分析中很难检测到低丰度的次要颗粒。我们开发了一种病毒颗粒分析方法,该方法整合了高灵敏度的分裂荧光素酶系统和密度梯度离心。本方案能够对表达日本脑炎病毒因子的细胞产生的颗粒进行高分辨率分析。