Expression of Membrane Proteins in Drosophila Melanogaster S2 Cells: Production and Analysis of a EGFP-Fused G Protein-Coupled Receptor as a Model
Expression of Membrane Proteins in Drosophila Melanogaster S2 Cells: Production and Analysis of a EGFP-Fused G Protein-Coupled Receptor as a Model
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DOI：
10.1007/978-1-60761-344-2_8
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发表时间：
2010-01-01
期刊：
影响因子：
--
通讯作者：
Wagner, Renaud
中科院分区：
文献类型：
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作者：
Brillet, Karl;Pereira, Carlos A.;Wagner, Renaud
在为功能性真核膜蛋白的异源表达选择合适宿主的过程中，果蝇S2细胞虽然尚未得到充分的探索，但似乎是哺乳动物细胞系或其他病毒感染昆虫细胞系统的有价值的替代品。这种非裂解的、基于质粒的系统实际上结合了几个主要的生理和生物过程优势，使其成为一种极具潜力和可扩展的细胞工具，用于各种应用的膜蛋白生产，诱导功能表征、药理学分析、分子模拟、结构分析或产生疫苗。本文以增强型绿色荧光蛋白(EGFP)/G蛋白偶联受体（EGFP- gpcr）为模型，提出了一系列有助于在S2细胞中成功表达膜蛋白的方案和提示。
In the process of selecting an appropriate host for the heterologous expression of functional eukaryotic membrane proteins, Drosophila S2 cells, although not yet fully explored, appear as a valuable alternative to mammalian cell lines or other virus-infected insect cell systems. This nonlytic, plasmid-based system actually combines several major physiological and bioprocess advantages that make it a highly potential and scalable cellular tool for the production of membrane proteins in a variety of applications, inducing functional characterization, pharmacological profiling, molecular simulations, structural analyses, or generation of vaccines. We present here a series of protocols and hints that would serve the successful expression of membrane proteins in S2 cells, using an enhanced green fluorescent protein (EGFP)/G protein-coupled receptor (EGFP-GPCR) as a model.