Lackner Andreas, Kreidl Emanuel, Peter-Vörösmarty Barbara, Spiegl-Kreinecker Sabine, Berger Walter, Grusch Michael
Department of Medicine I, Institute of Cancer Research, Medical University of Vienna, Vienna, Austria.
Methods Mol Biol. 2012;801:75-92. doi: 10.1007/978-1-61779-352-3_6.
The baculovirus Autographa californica multiple nuclear polyhedrosis virus (AcMNPV) has been widely used in biotechnology for protein expression in insect cells. Baculoviruses use arthropods as their natural hosts and are unable to replicate in mammalian cells. However, AcMNPV is able to enter many mammalian cell types and can be used for transgene expression if engineered to contain suitable expression cassettes. In this chapter, we describe the construction and application of a recombinant baculovirus containing a bicistronic expression cassette that can be used for stable protein expression in mammalian cells. As an example, the generation of glioblastoma and hepatocellular carcinoma cell lines stably expressing green fluorescent protein after puromycin selection is shown.
杆状病毒苜蓿银纹夜蛾多核型多角体病毒（AcMNPV）已在生物技术中广泛用于昆虫细胞中的蛋白质表达。杆状病毒以节肢动物为天然宿主，无法在哺乳动物细胞中复制。然而，AcMNPV能够进入多种哺乳动物细胞类型，如果经过工程改造含有合适的表达盒，就可用于转基因表达。在本章中，我们描述了一种含有双顺反子表达盒的重组杆状病毒的构建和应用，该病毒可用于哺乳动物细胞中的稳定蛋白质表达。作为一个例子，展示了在嘌呤霉素筛选后稳定表达绿色荧光蛋白的胶质母细胞瘤和肝癌细胞系的产生。
