Production of CCHF virus-like particle by a baculovirus-insect cell expression system

Virol Sin. 2011 Oct;26(5):338-46. doi: 10.1007/s12250-011-3209-6. Epub 2011 Oct 7.

Abstract

Crimean-Congo Haemorrhagic Fever Virus (CCHFV) is a tick-born virus of the Nairovirus genus within the Bunyaviridae family, which is widespread and causes high fatality. The nucleocapsid of CCHFV is comprised of N proteins that are encoded by the S segment. In this research, the N protein of CCHFV was expressed in insect cells using a recombinant baculovirus. Under an electron microscope, Virus-Like Particles (VLPs) with various size and morphology were observed in cytoplasmic vesicles in the infected cells. Sucrose-gradient purification of the cell lysate indicated that the VLPs were mainly located in the upper fraction after ultracentrifugation, which was confirmed by Western blot analysis and immuno-electron microscopy (IEM).

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Blotting, Western
  • Cell Line
  • Centrifugation, Density Gradient / methods
  • Gene Expression
  • Genetic Vectors*
  • Hemorrhagic Fever Virus, Crimean-Congo / genetics*
  • Microscopy, Electron, Transmission
  • Nucleocapsid Proteins / genetics*
  • Nucleocapsid Proteins / metabolism*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Spodoptera
  • Virosomes / genetics*
  • Virosomes / isolation & purification
  • Virosomes / metabolism*
  • Virosomes / ultrastructure

Substances

  • Nucleocapsid Proteins
  • Recombinant Proteins
  • Virosomes