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

被引:23
|
作者
Saito, T
Dojima, T
Toriyama, M
Park, EY
机构
[1] Shizuoka Univ, Fac Agr, Dept Appl Biol Chem, Shizuoka 4228529, Japan
[2] Kaken Pharmaceut Co Ltd, Prod Technol Lab, Fujieda, Shizuoka 4268646, Japan
关键词
cell cycle; Sf9; cell; baculovirus-insect cell expression system; GFPuv;
D O I
10.1016/S0168-1656(01)00398-4
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:121 / 129
页数:9
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