Snapp Erik Lee, Lajoie Patrick
Cold Spring Harb Protoc. 2011 Nov 1;2011(11):1368-9. doi: 10.1101/pdb.prot066571.
Eukaryotic cells are composed of an intricate system of internal membranes that are organized into different compartments--including the endoplasmic reticulum (ER), the nuclear envelope, the Golgi complex (GC), lysosomes, endosomes, caveolae, mitochondria, and peroxisomes--that perform specialized tasks within the cell. The localization and dynamics of intracellular compartments are now being studied in living cells because of the availability of green fluorescent protein (GFP)-fusion proteins and recent advances in fluorescent microscope imaging systems, such as the confocal laser-scanning microscope (CLSM). This protocol describes the steps for activating one of the first photoactivatable proteins, PA-GFP.
真核细胞由一个复杂的内膜系统组成，这些内膜被组织成不同的区室，包括内质网（ER）、核膜、高尔基体复合体（GC）、溶酶体、内体、小窝、线粒体和过氧化物酶体，它们在细胞内执行特定任务。由于绿色荧光蛋白（GFP）融合蛋白的可用性以及荧光显微镜成像系统（如共聚焦激光扫描显微镜（CLSM））的最新进展，目前正在活细胞中研究细胞内区室的定位和动态变化。本方案描述了激活首批光激活蛋白之一PA-GFP的步骤。
