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Efficient gene delivery into mammalian cells by recombinant baculovirus containing a hybrid cytomegalovirus promoter/Semliki Forest virus replicon
Author(s) -
Pan Yongfei,
Zhao Qian,
Fang Liurong,
Luo Rui,
Chen Huanchun,
Xiao Shaobo
Publication year - 2009
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.1390
Subject(s) - semliki forest virus , transduction (biophysics) , biology , reporter gene , replicon , microbiology and biotechnology , recombinant dna , gene delivery , transfection , alphavirus , green fluorescent protein , recombinant virus , virology , internal ribosome entry site , viral vector , gene , gene expression , virus , plasmid , translation (biology) , genetics , rna , biochemistry , messenger rna
Background Baculovirus, which is widely utilized as an excellent tool for the production of recombinant protein in insect cells, has recently emerged as a novel and attractive gene delivery vehicle for mammalian cells. Alphavirus, such as Semliki Forest virus (SFV), has also received considerable attention for use as expression vectors because of its self‐replicating property. In the present study, we investigated the characterization of recombinant baculovirus incorporating a hybrid cytomegalovirus (CMV) promoter/SFV replicon. Methods Recombinant baculovirus containing the hybrid CMV promoter/SFV replicon was constructed. Using enhanced green fluorescence protein (EGFP) as the reporter gene, gene delivery efficiencies and the ability to express heterogenous protein in mammalian cells were evaluated. Optimal transduction conditions, including transduction temperature, time and dose, were also investigated. Results The obtained recombinant baculovirus, Bac‐CMV/SFV‐EGFP, exhibited high transduction efficiency and high‐level expression of reporter protein in mammalian cells. Furthermore, this recombinant baculovirus could induce apoptosis in mammalian cells in the course of transduction, as demonstrated by the observed DNA laddering patterns and increased caspase‐3 activity. Conclusions The developed baculovirus vector has a high transduction efficiency and the ability to mediate foreign gene expression in mammalian cells. Taken together with its pro‐apoptotic properties, this baculovirus vector may provide an alternative tool for vaccine development. Copyright © 2009 John Wiley & Sons, Ltd.