A variety of species have evolved the ability to emit light biochemically. Among those that have found the most use in molecular biology applications are the North American firefly photinus pyralis, the marine copepod Gaussia princeps and the bacterium Vibrio harveyi. The firefly luciferase gene (luc) has been cloned into a variety of species, becoming a popular marker gene for plants, bacteria and mammalian cells. Measurement of transciptional regulation by coupling the expression of a reporter gene is widely used to monitor a variety of physiological events including the action of specific receptor elements on gene expression, signal transduction, expression of transcription factors, protein-protein interactions, viral infection and processing. The firefly luciferase gene encodes a functional protein immediately upon transcription, and an assay for the resulting light production is highly sensitive (very low background), rapid, and reliable, making luciferase a popular choice for reporter cloning experiments. These assays are also well suited to automation and high-throughput applications. The substrate, coelenterazine is common to a number of marine bioluminescent reactions, including those from Renilla, Gaussia, Aequorea and Watesenia. In some of these reactions it is utilized as a simple substrate being catalytically turned over in the bioluminescent reaction, while in others, such as in the photoprotein systems of Mneiopsis, it is incorporated as part of the photoprotein. The substrate h-coelenterate luciferin is a coelenterazine luciferin analog wherein one of the phenolic groups is replaced with a proton. This substrate exhibits a high recharging efficiency and even higher light output., fluorometric, and spectrophotometric measurements of β-glucuronidase gene fusion expression.
This kit contains the all of the reagents including the primary substrate of the firefly luciferase light producing system, D-luciferin, as well as the lysis and luciferase enzyme assay buffers. Learn More