lacZ Reporter System as a Tool to Study Virulence Gene Regulation in Bacterial Pathogens
- 1.8k Downloads
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 .
Key wordsBeta-galactosidase (β-gal) lacZ Lactose operon O-nitrophenyl-β-d-galactoside (ONPG) Reporter Gene regulation
The research in Gal-Mor lab is supported by a grant number 1096.39.11/2010 from the German-Israeli Foundation for Scientific Research and Development (GIF); by a grant number 999/14 from the Israel Science Foundation (ISF) and by grant number 3-0000-12435 from Infect-ERA and the Chief Scientist’s Bureau in the Israeli Ministry of Health.
- 2.Jeffrey MH (1972) In: experiments in molecular genetics. Cold Spring Harbor Laboratories, Cold Spring Harbor, NY, pp 352–355Google Scholar
- 3.Casadaban MJ, Chou J, Cohen SN (1980) In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol 143:971–980PubMedPubMedCentralGoogle Scholar