Facultad de Biología, Departamento de Genética, Universidad de Sevilla, Sevilla, Spain.
Methods Mol Biol. 2021;2182:141-151. doi: 10.1007/978-1-0716-0791-6_13.
The luxCDABE operon of Photorhabdus luminescens can be used as a bioluminescent reporter to measure gene transcription nondestructively. Here we describe protocols to (1) generate random transcriptional fusions of the lux operon to genes of the Salmonella genome, (2) screen for specific fusions with constitutive expression, Salmonella pathogenicity island 1-related expression, or Salmonella pathogenicity island 2-related expression, and (3) determine the site of luxCDABE integration.
发光杆菌的 luxCDABE 操纵子可用作生物发光报告基因，非破坏性地测量基因转录。本文介绍了以下方案：（1）生成 lux 操纵子与沙门氏菌基因组基因的随机转录融合；（2）筛选具有组成型表达、与沙门氏菌致病岛 1 相关表达或与沙门氏菌致病岛 2 相关表达的特定融合子；（3）确定 luxCDABE 整合的位置。
