Formation of peste des petits ruminants spikeless virus-like particles by co-expression of M and N proteins in insect cells

Res Vet Sci. 2014 Feb;96(1):213-6. doi: 10.1016/j.rvsc.2013.10.012. Epub 2013 Nov 7.

Abstract

Peste des petits ruminants virus (PPRV) has a non-segmented negative sense RNA genome and is classified within the Morbillivirus genus of the Paramyxoviridae. Using the Bac-to-Bac® baculovirus expression system, we constructed recombinant baculoviruses that were able to co-express the PPRV matrix and nucleocapsid proteins in insect cells under the control of the polyhedron and p10 promoters, respectively. The results showed that although both structural proteins were expressed at a relatively low level, the interaction between them caused the formation of virus-like particles (VLPs) by viewing of transmission electron microscopy. The VLPs morphologically resembled authentic PPRVs but lacked spikes protruding from the particulate surfaces. Interestingly, the diameter of PPRV VLPs ranged from 100 to 150 nm, far less than the mean diameter (400-500 nm) of parental virions.

Keywords: Baculovirus expression system; Co-expression; Matrix protein; Nucleocapsid protein; Peste des petits ruminants virus; Virus-like particle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Cloning, Molecular / methods
  • Microscopy, Electron, Transmission
  • Nucleocapsid Proteins / genetics*
  • Peste-des-Petits-Ruminants / virology*
  • Peste-des-petits-ruminants virus / genetics*
  • Peste-des-petits-ruminants virus / ultrastructure
  • Recombinant Proteins / genetics*
  • Recombinant Proteins / ultrastructure
  • Viral Matrix Proteins / genetics*

Substances

  • Nucleocapsid Proteins
  • Recombinant Proteins
  • Viral Matrix Proteins