See All pGL4 Vectors
Creating a cell line with an indicator of a functional signaling pathway is useful for deciphering the components in a signaling pathway. These tools are made by insertion of multiple repeats of a response element upstream of a minimal promoter (minP). Promega has designed vectors that report the activity of a variety of pathways using the optimized luc2 firefly luciferase gene in the pGL4 backbone. These vectors also have a hygromycin resistance selectable marker, allowing use either in transient transfection experiments or for selection of a stable cell line.
Also available for construction of pathway reporters are minimal promoter (minP) vectors with three varieties of engineered firefly luciferase genes: luc2, luc2P orluc2CP. The luc2 gene is engineered to remove most cryptic transcription factor binding sites and improve mammalian expression through codon optimization. Theluc2P and luc2CP and RapidResponse™ genes are luc2 genes appended with degradation sequences to influence the cellular half-life of the luc2 gene. The RapidResponse™ genes respond more rapidly to stimuli but at the expense of signal intensity. The luc2P (1-hour half-life) gene responds more rapidly than luc2 (3-hour half-life) with moderate signal intensity, and the luc2CP (0.4-hour half-life) responds more quickly with the lowest signal intensity. The minP vectors are available with or without selectable markers (hygromycin). To speed research, several pre-designed response element vectors are available already assembled in the pGL4.27 Vector. Some of these also are available stable cell lines (GloResponse™ Cell Lines).
Features - Benefits
Pre-designed vectors remove the need to clone and validate an assay.
Increased Reporter Gene Expression: Codon optimization of synthetic genes for mammalian expression.
Reduced Background and Risk of Expression Artifacts: Removal of cryptic DNA regulatory elements and transcription factor binding sites.
Improved Temporal Response: Rapid Response™ technology using destabilized luciferase genes.
Easy Transition from Transient to Stable Cells: Choice of mammalian selectable markers.