Institute of Biotechnology, National Cheng Kung University, No. 1, University Road, Tainan 701, Taiwan, ROC.
Biochem Biophys Res Commun. 2011 Feb 11;405(2):297-302. doi: 10.1016/j.bbrc.2011.01.032. Epub 2011 Jan 8.
The baculovirus group of insect viruses is widely used for foreign gene introduction into mammalian cells for gene expression and protein production; however, the efficiency of baculovirus entry into mammalian cells is in general still low. In this study, two recombinant baculoviruses were engineered and their ability to improve viral entry was examined: (1) cytoplasmic transduction peptide (CTP) was fused with baculovirus envelope protein, GP64, to produce a cytoplasmic membrane penetrating baculovirus (vE-CTP); and (2) the protein transduction domain (PTD) of HIV TAT protein was fused with the baculovirus capsid protein VP39 to form a nuclear membrane penetrating baculovirus (vE-PTD). Transduction experiments showed that both viruses had better transduction efficiency than vE, a control virus that only expresses EGFP in mammalian cells. Interestingly, vE-CTP and vE-PTD were also able to improve the transduction efficiency of a co-transduced baculovirus, resulting in higher levels of gene expression. Our results have described new routes to further enhance the development of baculovirus as a tool for gene delivery into mammalian cells.
杆状病毒科的昆虫病毒被广泛用于将外源基因导入哺乳动物细胞中进行基因表达和蛋白质生产；然而，杆状病毒进入哺乳动物细胞的效率通常仍然较低。在本研究中，构建了两种重组杆状病毒，并研究了它们提高病毒进入效率的能力：（1）将细胞质转导肽（CTP）与杆状病毒包膜蛋白 GP64 融合，产生细胞质膜穿透杆状病毒（vE-CTP）；（2）将 HIV TAT 蛋白的蛋白转导结构域（PTD）与杆状病毒衣壳蛋白 VP39 融合，形成核膜穿透杆状病毒（vE-PTD）。转导实验表明，与仅在哺乳动物细胞中表达 EGFP 的对照病毒 vE 相比，这两种病毒都具有更好的转导效率。有趣的是，vE-CTP 和 vE-PTD 还能够提高共转导杆状病毒的转导效率，从而提高基因表达水平。我们的研究结果描述了进一步增强杆状病毒作为基因传递到哺乳动物细胞的工具的新途径。
