A baculovirus superinfection system:: Efficient vehicle for gene transfer into Drosophila S2 cells

被引:47
|
作者
Lee, DF
Chen, CC
Hsu, TA
Juang, JL
机构
[1] Natl Hlth Res Inst, Div Mol & Genom Med, Taipei 11529, Taiwan
[2] Natl Hlth Res Inst, Div Biotechnol & Pharmaceut Res, Taipei 11529, Taiwan
关键词
D O I
10.1128/JVI.74.24.11873-11880.2000
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The baculovirus expression vector system is considered to be a safe, powerful, but cell-lytic heterologous protein expression system in insect cells. We show here that there is a new baculovirus system for efficient gene transfer and expression using the popular and genetically well-understood Drosophila S2 cells. The recombinant baculovirus was constructed to carry an enhanced green fluorescent protein under the control of polyhedrin promoter as a fluorescent selection marker in the Sf21 cell line. Recombinant baculoviruses were then used to transduce S2 cells with target gene expression cassettes containing a Drosophila heat shock protein 70, an actin 5C, or a metallothionein promoter. Nearly 100% of the S2 cells showed evidence of gene expression after infection. The time course for the optimal protein expression peaked at 24 to 36 h postinfection, which is significantly earlier than a polyhedrin-driven protein expression in Sf21 cells. Importantly, S2 cells did not appear to be lysed after infection, and the protein expression levels are comparable to those of proteins under the control of polyhedrin promoter in several lepidopteran cell lines. Most surprisingly, S2 cells permit repetitive infections of multiple baculoviruses over time. These findings clearly suggest that this baculovirus-S2 system may effect the efficient gene transfer and expression system of the well-characterized Drosophila S2 cells.
引用
收藏
页码:11873 / 11880
页数:8
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