Derbise Anne, Dussurget Olivier, Carniel Elisabeth, Pizarro-Cerdá Javier
Yersinia Research Unit/Yersinia National Reference Laboratory/WHO Collaborating Center, Institut Pasteur, Paris, France.
Université de Paris, Sorbonne Paris Cité, Paris, France.
Methods Mol Biol. 2019;2010:85-97. doi: 10.1007/978-1-4939-9541-7_7.
Bioluminescence imaging (BLI) has become a major strategy for real-time analysis of dynamic biological processes. In particular, bioluminescent reporter microorganisms have been designed to advance our understanding of infectious diseases. Non-invasive monitoring of light-emitting pathogenic bacteria has revealed novel features of pathogenesis and enabled quantitative and qualitative analysis of antibacterial therapies. Transcriptional gene fusions using the bacterial luciferase operon luxCDABE as a reporter have been successfully used to monitor gene expression in vitro and in vivo, leading to valuable applications and major findings. In this chapter, we describe the construction of Yersinia pestis strains bearing a chromosomal copy of the luxCDABE operon under the control of promoters regulated by temperature and their application to quantify gene expression in real-time in bacteria growing in vitro and in a murine bubonic plague model.
生物发光成像（BLI）已成为实时分析动态生物学过程的主要策略。特别是，已设计出生物发光报告微生物以增进我们对传染病的理解。对发光病原菌的非侵入性监测揭示了发病机制的新特征，并实现了对抗菌疗法的定量和定性分析。使用细菌荧光素酶操纵子luxCDABE作为报告基因的转录基因融合已成功用于监测体外和体内的基因表达，从而带来了有价值的应用和重大发现。在本章中，我们描述了构建在温度调控的启动子控制下带有luxCDABE操纵子染色体拷贝的鼠疫耶尔森菌菌株，以及它们在体外生长的细菌和小鼠腺鼠疫模型中实时定量基因表达的应用。
