Ray Pritha, Gambhir Sanjiv Sam
Molecular Imaging Program at Stanford, Department of Radiology, Bio-X Program, Stanford University, USA.
Methods Mol Biol. 2007;411:131-44. doi: 10.1007/978-1-59745-549-7_10.
Bioluminescence imaging has become a very popular tool for noninvasive monitoring of fundamental biological and molecular processes in small living subjects. Luciferases are light-emitting enzymes that can generate light (known as bioluminescence) after reacting with specific substrates. The emitted light is used as a detection system for luciferase activity, which acts as a "reporter" for the activity of any regulatory elements that control its expression. These enzymes are isolated from various organisms, conveniently modified for expression in mammalian cells, and are extensively used in molecular biology and cell culture experiments. Recent advances in optical technology have opened a new dimension for in vivo application of luciferase enzymes in biomedical research. The most commonly utilized luciferases for in vivo bioluminescence are isolated from two very different sources: firefly luciferase (or beetle luciferase) and renilla luciferase (isolated from sea pansy). Although both these luciferases can produce light following interaction with the substrates, structurally and biochemically they are very different. Here we describe the methods and applications of firefly and renilla luciferases in molecular imaging using small animals.
生物发光成像已成为一种非常流行的工具，用于对小型活体动物体内的基本生物学和分子过程进行无创监测。荧光素酶是一种发光酶，与特定底物反应后可产生光（即生物发光）。发出的光用作荧光素酶活性的检测系统，而荧光素酶活性则作为控制其表达的任何调控元件活性的“报告分子”。这些酶从各种生物体中分离出来，经过方便的修饰后可在哺乳动物细胞中表达，并广泛应用于分子生物学和细胞培养实验。光学技术的最新进展为荧光素酶在生物医学研究中的体内应用开辟了新的维度。用于体内生物发光的最常用荧光素酶来自两个非常不同的来源：萤火虫荧光素酶（或甲虫荧光素酶）和海肾荧光素酶（从海鳃中分离）。尽管这两种荧光素酶与底物相互作用后都能产生光，但它们在结构和生化性质上有很大差异。在这里，我们描述萤火虫和海肾荧光素酶在使用小动物进行分子成像中的方法和应用。
