Wiles Siouxsie, Robertson Brian D, Frankel Gad, Kerton Angela
Department of Infectious Diseases and Immunity, Imperial College London, London, UK.
Methods Mol Biol. 2009;574:137-53. doi: 10.1007/978-1-60327-321-3_12.
Bioluminescence is an excellent reporter system for analysing bacterial colonization and clearance dynamics in vivo. Many bacterial species have been rendered bioluminescent, allowing the sensitive detection of bacterial burden and metabolic activity in real-time and in situ in living animals. In this chapter we describe the protocols for characterizing in vivo infection models using bioluminescent bacteria: from real-time imaging in living animals by bioluminescence imaging (BLI) to ex vivo BLI of harvested organs and tissues and, finally, to quantification of bacterial numbers in organ and tissue homogenates by luminometry and viable counts. While the lux operon from Photorhabdus luminescens is ideally suited for use in such models, there may be times when alternative luciferases, such as those from the firefly (luc) or marine copepods (Gluc), may be more appropriate. Here we describe the protocols required to monitor colonization and clearance dynamics using bioluminescent bacteria that are lux-, luc-, or Gluc-positive.
生物发光是用于分析体内细菌定殖和清除动态的优秀报告系统。许多细菌物种已被改造为具有生物发光能力，从而能够在活体动物中实时、原位灵敏地检测细菌负荷和代谢活性。在本章中，我们描述了使用生物发光细菌表征体内感染模型的方案：从通过生物发光成像（BLI）对活体动物进行实时成像，到对收获的器官和组织进行离体BLI，最后通过发光测定法和活菌计数对器官和组织匀浆中的细菌数量进行定量。虽然来自发光杆菌属的lux操纵子非常适合用于此类模型，但有时其他荧光素酶，如来自萤火虫（luc）或海洋桡足类动物（Gluc）的荧光素酶，可能更合适。在这里，我们描述了使用lux、luc或Gluc阳性的生物发光细菌监测定殖和清除动态所需的方案。
