Bio-Protocol (May 2020)

Split Nano Luciferase-based Assay to Measure Assembly of Japanese Encephalitis Virus

  • Simon Goto,
  • Kotaro Ishida,
  • Ryosuke Suzuki,
  • Eiji Morita

DOI
https://doi.org/10.21769/BioProtoc.3606
Journal volume & issue
Vol. 10, no. 9

Abstract

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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.