Use of Bacterial Luciferase as a Reporter Gene in Eukaryotic Systems.

Reporter gene assays are powerful tools for monitoring dynamic molecular changes and for evaluating the responses that occur at the genetic elements within cells in response to exogenous molecules. In general, various protein systems can be used as reporter genes, including luciferases. Here, the present protocol introduces a unique reporter gene system for monitoring molecular events in cells using bacterial luciferase (lux), which can generate blue-green light suitable for gene reporter applications with the highest cost performance. The protocol also guides the assay conditions and necessary components for using of lux gene (lux) as a eukaryotic reporter system. The lux system can be applied to monitor variety of molecular events inside mammalian cellular systems.

Bioluminescent monitoring of detoxification processes: activity of humic substances in quinone solutions.

This study deals with application of bioluminescent assay systems to evaluate the detoxifying effect of humic substances (HS) on the solutions of organic oxidizers - quinones. A series of homologous quinones with different redox characteristics: 1,4-benzoquinone, tetrafluoro-1,4-benzoquinone, methyl-1,4-benzoquinone, tetramethyl-1,4-benzoquinone, and 1,4-naphtoquinone, was used. Bioluminescent bacteria Photobacterium phosphoreum, and NADH:FMN-oxidoreductase-luciferase enzyme system isolated from these bacteria were used as assay systems. The toxicity was compared in the presence and in the absence of HS. Variation of complexity of bioassays (in vivo or in vitro) combined with spectrometric and microscopic methods, provides insight into the process of detoxification in quinone solutions. Two ways of HS effect were studied: the reduction activity of HS and intensification of self-protection of bacterial cells on HS addition.