Yang Jianying, Reth Michael
Faculty of Biology, BIOSS Centre for Biological Signalling Studies, University of Freiburg, Max-Planck-Institute of Immunobiology and Epigenetics, Freiburg, Germany.
Methods Mol Biol. 2012;813:331-41. doi: 10.1007/978-1-61779-412-4_20.
Synthetic biology is an engineering approach to biology. A synthetic biologist wants to describe biological molecules and their subdomains as well-defined parts of a molecular machine. To achieve this goal, synthetic biologists rebuild minimal functional biological systems from well-defined parts or they design new molecules that do not exist in nature but have new and useful functions. In short, these engineering approaches can be summarized as "rebuild, alter, and understand." The Drosophila S2 Schneider cell is a useful tool for both rebuilding and redesigning approaches. S2 cells are phagocytic cells that easily take up large amounts of DNA from the cell culture. They, thus, have a high cotransfection rate, allowing the coexpression of up to 12 different proteins. We have developed a transient transfection protocol allowing the rapid and parallel analysis of wild-type and altered forms of a biological system. This chapter describes our methods to rebuild and better understand mammalian signaling systems in the evolutionary distant environment of Drosophila S2 cells.
合成生物学是一种生物学的工程学方法。合成生物学家希望将生物分子及其亚结构域描述为分子机器中定义明确的部件。为实现这一目标,合成生物学家从定义明确的部件重建最小功能生物系统,或者设计自然界中不存在但具有新的有用功能的新分子。简而言之,这些工程学方法可以概括为“重建、改变和理解”。果蝇S2 Schneider细胞是重建和重新设计方法的有用工具。S2细胞是吞噬细胞,能轻易从细胞培养物中摄取大量DNA。因此,它们具有高共转染率,允许同时表达多达12种不同的蛋白质。我们开发了一种瞬时转染方案,可对生物系统的野生型和改变形式进行快速并行分析。本章描述了我们在果蝇S2细胞这个进化距离较远的环境中重建和更好理解哺乳动物信号系统的方法。