Aviv Gili, Gal-Mor Ohad
The Infectious Diseases Research Laboratory, Sheba Medical Center, Tel-Hashomer, Ramat Gan, Israel.
Department of Clinical Microbiology and Immunology, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
Methods Mol Biol. 2018;1734:39-45. doi: 10.1007/978-1-4939-7604-1_5.
β-galactosidase assay has been established as one of the most widely used reporters and can be effectually exploited to study promoter activity of Salmonella and other pathogens under various conditions. This method includes a preliminary stage of fusing the target promoter to a promoter-less lacZ gene encoding for the enzyme β-galactosidase. Supplementation of the synthetic ONPG substrate results in the accumulation of a chromogenic product proportionally to the activity of the fused promoter. Here we demonstrate the usage of this reporter system to study the regulation of the Salmonella Type three secretion system effector gene sseL in S. Typhimurium [1].
β-半乳糖苷酶检测已成为应用最为广泛的报告基因检测方法之一，可有效用于研究沙门氏菌及其他病原体在各种条件下的启动子活性。该方法包括一个初步阶段，即将目标启动子与编码β-半乳糖苷酶的无启动子lacZ基因融合。添加合成的ONPG底物后，会产生一种与融合启动子活性成比例的显色产物。在此，我们展示了该报告系统用于研究鼠伤寒沙门氏菌中沙门氏菌Ⅲ型分泌系统效应基因sseL调控的应用[1]。
