New luciferase vectors for studying signal transduction pathways
Li Xu Tim Sanchez Chao-Feng Zheng
Stratagene Cloning Systems, Inc.
Enhancer elements are the convergent points for many intracellular signal transduction pathways. For simple, rapid and convenient assessment of the in vivo activation of these pathways, Stratagene has designed the PathDetect in vivo signal transduction pathway cis-reporting systems. Each of the cis-reporter plasmids contains the luciferase reporter gene driven by a basic promoter element (TATA box) joined to tandem repeats of AP-1, CRE, SRE, NF-kB or SRF binding elements. When a plasmid expressing the gene of interest is cotransfected into mammalian cells with a cis-reporter plasmid, increased luciferase activity indicates transcription activation. The PathDetect cis-reporting systems can be used for studying the in vivo effects of a new gene, growth factor or drug candidate on signaling pathways. The effects of extracellular stimuli can also be studied with these systems.
Extracellular signals trigger the sequential activation of a series of
intracellular signaling molecules, such as protein kinases and phosphatases.1,2
The final step in the activation of many pathways is the binding of an activated
transcription factor to specific enhancer elements found in the promoters of
various cellular genes. This binding modulates transcription of these genes. The
transcription level of cellular genes reflects the activation status of the
involved signaling events. Transcription and expression levels are typically
monitored by Northern and Western blotting. To determine the activation of the
promoter, and hence signaling events, we have designed vectors that use a
reporter gene in place of the cellular gene. The reporter gene encodes the
firefly luciferase enzyme that can be monitored ea