Nye Jonathan, Buster Daniel W, Rogers Gregory C
Department of Cellular & Molecular Medicine, Arizona Cancer Center, Room 3951, University of Arizona, 1515 N. Campbell Avenue, Tucson, AZ, 85724, USA.
Methods Mol Biol. 2014;1136:81-101. doi: 10.1007/978-1-4939-0329-0_6.
Cultured Drosophila cell lines have been developed into a powerful tool for studying a wide variety of cellular processes. Their ability to be easily and cheaply cultured as well as their susceptibility to protein knockdown via double-stranded RNA-mediated interference (RNAi) has made them the model system of choice for many researchers in the fields of cell biology and functional genomics. Here we describe basic techniques for gene knockdown, transgene expression, preparation for fluorescence microscopy, and centrosome enrichment using cultured Drosophila cells with an emphasis on studying the microtubule cytoskeleton.
培养的果蝇细胞系已发展成为研究多种细胞过程的强大工具。它们易于培养且成本低廉,以及通过双链RNA介导的干扰(RNAi)对蛋白质敲低的敏感性,使其成为细胞生物学和功能基因组学领域许多研究人员的首选模型系统。在这里,我们描述了使用培养的果蝇细胞进行基因敲低、转基因表达、荧光显微镜准备和中心体富集的基本技术,重点是研究微管细胞骨架。