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首页> 外文期刊>The journal of gene medicine >Efficient gene delivery into mammalian cells by recombinant baculovirus containing a hybrid cytomegalovirus promoter/Semliki Forest virus replicon
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Efficient gene delivery into mammalian cells by recombinant baculovirus containing a hybrid cytomegalovirus promoter/Semliki Forest virus replicon

机译：含有杆状巨细胞病毒启动子/塞姆利基森林病毒复制子的重组杆状病毒将基因有效地传递到哺乳动物细胞中

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Background Baculovirus, which is widely utilized as an excellent tool forthe production of recombinant protein in insect cells, has recently emerged asa novel and attractive gene delivery vehicle for mammalian cells. Alphavirus,such as Semliki Forest virus (SFV), has also received considerable attentionfor use as expression vectors because of its self-replicating property. Inthe present study, we investigated the characterization of recombinantbaculovirus incorporating a hybrid cytomegalovirus (CMV) promoter/SFVreplicon. Methods Recombinant baculovirus containing the hybrid CMV pro-moter/SFV replicon was constructed. Using enhanced green fluorescenceprotein (EGFP) as the reporter gene, gene delivery efficiencies and the abilityto express heterogenous protein in mammalian cells were evaluated. Optimaltransduction conditions, including transduction temperature, time and dose,were also investigated. Results The obtained recombinant baculovirus, Bac-CMV/SFV-EGFP, exhib-ited high transduction efficiency and high-level expression of reporter proteinin mammalian cells. Furthermore, this recombinant baculovirus could induceapoptosis in mammalian cells in the course of transduction, as demonstratedby the observed DNA laddering patterns and increased caspase-3 activity. Conclusions The developed baculovirus vector has a high transductionefficiency and the ability to mediate foreign gene expression in mammaliancells. Taken together with its pro-apoptotic properties, this baculovirus vectormay provide an alternative tool for vaccine development.
机译：背景技术杆状病毒已被广泛用作在昆虫细胞中产生重组蛋白的优良工具，最近已成为哺乳动物细胞的一种新颖且有吸引力的基因传递载体。诸如Semliki Forest病毒（SFV）之类的α病毒由于具有自我复制特性，因此作为表达载体也受到了广泛的关注。在本研究中，我们调查了重组杆状病毒的特征，该重组杆状病毒结合了巨细胞巨细胞病毒（CMV）启动子/ SFVreplicon。方法构建含有CMV启动子/ SFV复制子的重组杆状病毒。使用增强型绿色荧光蛋白（EGFP）作为报告基因，评估了基因传递效率和在哺乳动物细胞中表达异源蛋白的能力。还研究了最佳的转导条件，包括转导温度，时间和剂量。结果获得的重组杆状病毒Bac-CMV / SFV-EGFP在哺乳动物细胞中表现出较高的转导效率和报告蛋白的高水平表达。此外，该重组杆状病毒可在转导过程中诱导哺乳动物细胞凋亡，如观察到的DNA梯形图样和增加的caspase-3活性所证明。结论开发的杆状病毒载体具有很高的转导效率，并具有介导哺乳动物细胞中外源基因表达的能力。结合其促凋亡特性，这种杆状病毒载体可为疫苗开发提供替代工具。

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