Expression of rice gall dwarf virus outer coat protein gene (S8) in insect cells

Virol Sin. 2010 Dec;25(6):401-8. doi: 10.1007/s12250-010-3152-y. Epub 2010 Dec 21.

Abstract

To obtain the P8 protein of Rice gall dwarf virus (RGDV) with biological activity, its outer coat protein gene S8 was expressed in Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression system. The S8 gene was subcloned into the pFastBac™1 vector, to produce the recombinant baculovirus transfer vector pFB-S8. After transformation, pFB-S8 was introduced into the competent cells (E. coli DH10Bac) containing a shuttle vector, Bacmid, generating the recombinant bacmid rbpFB-S8. After being infected by recombinant baculovirus rvpFB-S8 at different multiplicities of infection, Sf9 cells were collected at different times and analyzed by SDS-PAGE, Western blotting and immunofluorescence microscopy. The expression level of the P8 protein was highest between 48-72 h after transfection of Sf9 cells. Immunofluorescence microscopy showed that P8 protein of RGDV formed punctate structures in the cytoplasm of Sf9 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Baculoviridae / genetics
  • Blotting, Western
  • Capsid Proteins / biosynthesis*
  • Capsid Proteins / genetics*
  • Cell Line
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Gene Expression*
  • Genetic Vectors
  • Microscopy, Fluorescence
  • Oryza
  • Reoviridae / genetics*
  • Spodoptera

Substances

  • Capsid Proteins