Department of Biotechnology, Institut fuer Biochemie, Biotechnologie und Bioinformatik, Technische Universitaet Braunschweig, Braunschweig, Germany.
RG Cellular Proteomics, Department of Structure and Function of Proteins, Helmholtz Zentrum für Infektionsforschung GmbH, Braunschweig, Germany.
Methods Mol Biol. 2021;2305:129-140. doi: 10.1007/978-1-0716-1406-8_6.
The expression of mammalian recombinant proteins in insect cell lines using transient-plasmid-based gene expression enables the production of high-quality protein samples. Here, the procedure for virus-free transient gene expression (TGE) in High Five insect cells is described in detail. The parameters that determine the efficiency and reproducibility of the method are presented in a robust protocol for easy implementation and set-up of the method. The applicability of the TGE method in High Five cells for proteomic, structural, and functional analysis of the expressed proteins is shown.
利用基于瞬时质粒的基因表达在昆虫细胞系中表达哺乳动物重组蛋白可生产高质量的蛋白质样品。本文详细描述了在 High Five 昆虫细胞中进行无病毒瞬时基因表达 (TGE) 的过程。该方法效率和重现性的决定因素以稳健的方案呈现，便于轻松实施和设置该方法。TGE 方法在 High Five 细胞中用于表达蛋白的蛋白质组学、结构和功能分析的适用性也得到了展示。
