The effect of cell cycle on GFPuv gene expression in the baculovirus expression system.
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.
Saito
Takehiko
T
Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
Dojima
Takashi
T
Toriyama
Masaru
M
Park
Enoch Y
EY
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics; D005455:Fluorescent Antibody Technique; D016195:G2 Phase; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000235:genetics; D018613:Microscopy, Confocal; D008938:Mitosis; D015739:Nocodazole / Q000494:pharmacology; D016196:S Phase; D018411:Spodoptera / Q000378:metabolism*; D014162:Transfection
D004268:DNA-Binding Proteins; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D015739:Nocodazole