Li Li, Wang Xin, Yin Juan, Zhong Jiang
Department of Microbiology and Microbial Engineering, School of Life Sciences, Fudan University, Shanghai 200433, China.
Sheng Wu Gong Cheng Xue Bao. 2009 Oct;25(10):1558-63.
In order to improve the transduction efficiency of insect baculovirus in mammalian cells, we constructed two recombinant baculoviruses, AcRed-tat and AcRed. Both viruses expressed red fluorescence protein gene (dsRed) as a reporter in mammalian cell lines. AcRed-tat also contained the coding sequence of HIV-1 Tat transduction peptide fused with viral major capsid protein gene vp39 and enhanced green fluorescence gene (egfp) driven by virus polyhedrin promoter. It expressed the Tat fusion protein in infected insect cells, which was incorporated into the nucleocapsids of progeny virus. As a control, AcRed had the fusion gene of vp39 and egfp driven by polyhedrin promoter. Flow cytometry analysis demonstrated that although similar level of red fluorescence was produced in HEK23 cells transduced by the two recombinant viruses, significantly higher red fluorescence level was seen in CHO and Vero cells transduced by AcRed-tat than that by AcRed. These results suggested that Tat transduction peptide might improve the baculovirus-mediated gene expression in some mammalian cells. Our work provided a new approach to improve baculovirus as a gene delivery vector for mammalian cells.
为了提高昆虫杆状病毒在哺乳动物细胞中的转导效率，我们构建了两种重组杆状病毒，AcRed-tat和AcRed。两种病毒均表达红色荧光蛋白基因（dsRed）作为哺乳动物细胞系中的报告基因。AcRed-tat还包含与病毒主要衣壳蛋白基因vp39融合的HIV-1 Tat转导肽的编码序列以及由病毒多角体蛋白启动子驱动的增强型绿色荧光基因（egfp）。它在受感染的昆虫细胞中表达Tat融合蛋白，该蛋白被整合到子代病毒的核衣壳中。作为对照，AcRed具有由多角体蛋白启动子驱动的vp39和egfp融合基因。流式细胞术分析表明，尽管两种重组病毒转导的HEK293细胞产生的红色荧光水平相似，但AcRed-tat转导的CHO和Vero细胞中的红色荧光水平明显高于AcRed转导的细胞。这些结果表明，Tat转导肽可能会提高杆状病毒介导的某些哺乳动物细胞中的基因表达。我们的工作为改进杆状病毒作为哺乳动物细胞基因递送载体提供了一种新方法。
