Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, PR China.
Key Laboratory of Microbiology, College of Life Science, Heilongjiang University, Harbin 150080, PR China.
J Biotechnol. 2014 Mar 10;173:41-6. doi: 10.1016/j.jbiotec.2014.01.003. Epub 2014 Jan 18.
The baculovirus gene expression system is an efficient and safe protein expression system, since baculoviruses cannot replicate in mammalian cells. In order to improve the transduction efficiency and increase the reporter gene expression levels delivered by baculoviruses, we tested in the baculovirus expression cassette the Woodchuck hepatitis virus response element (WPRE), and AAV-derived inverted terminal repeats (ITRs) and the truncated vesicular stomatitis virus G protein (VSV-GED). The results showed that WPRE and VSV-GED have synergistic effects and could enhance the expression efficiency of enhanced green fluorescence protein (eGFP), and that ITRs effectively extended the duration of eGFP expression. We also demonstrated that the efficiency of eGFP expression varied under the control of the CMV, CBA, EF1-α or WSSV ie1 promoters in different cell lines.
杆状病毒基因表达系统是一种高效、安全的蛋白表达系统，因为杆状病毒不能在哺乳动物细胞中复制。为了提高转导效率并增加杆状病毒递送的报告基因表达水平，我们在杆状病毒表达盒中测试了 Woodchuck 肝炎病毒反应元件（WPRE）、AAV 衍生的反向末端重复序列（ITRs）和截短的水疱性口炎病毒 G 蛋白（VSV-GED）。结果表明，WPRE 和 VSV-GED 具有协同作用，可以增强增强型绿色荧光蛋白（eGFP）的表达效率，而 ITRs 则有效地延长了 eGFP 表达的持续时间。我们还证明，eGFP 的表达效率在不同细胞系中受 CMV、CBA、EF1-α 或 WSSV ie1 启动子的控制而有所不同。
