The effect of cell cycle on GFPuv gene expression in the baculovirus expression system

J Biotechnol. 2002 Feb 14;93(2):121-9. doi: 10.1016/s0168-1656(01)00398-4.

Abstract

The effects of cell cycle on recombinant protein production and infection yield in the baculovirus-insect cell expression system (BES) were investigated. When, at any cell cycle phase, the host cell was infected by baculovirus, the cell cycle was finally arrested at the S or G(2)/M phase with 4n DNA. In the case of G(1) or S phase-infection, cell cycle of virus-infected cells began to be arrested at S phase from 8 h post-infection or at G(2)/M phase from 4 h post-infection, respectively; while, in the case of M phase-infection, cell cycle was arrested at S phase after 12 h post-infection. When the host cell was infected at the G(1) phase, average intracellular GFPuv fluorescence intensity was 1.3-fold higher than that at G(2)/M phase at 24 h post-infection. The GFPuv expression corresponded to the profile of the G(1) cell cycle in the BES. Infection yield was measured by detection of intracellular DNA binding protein using immunohistochemical method within 7 h post-infection. The infection yield at G(1) or S phase-infection was 1.5-1.8-fold higher than that at G(2)/M phase-infection.

MeSH terms

  • Animals
  • Baculoviridae / genetics*
  • Cell Cycle* / drug effects
  • Cell Line
  • DNA-Binding Proteins / analysis
  • DNA-Binding Proteins / genetics
  • Fluorescent Antibody Technique
  • G2 Phase
  • Gene Expression*
  • Genetic Vectors
  • Green Fluorescent Proteins
  • Immunohistochemistry
  • Luminescent Proteins / genetics
  • Microscopy, Confocal
  • Mitosis
  • Nocodazole / pharmacology
  • S Phase
  • Spodoptera / metabolism*
  • Transfection

Substances

  • DNA-Binding Proteins
  • Luminescent Proteins
  • Green Fluorescent Proteins
  • Nocodazole