pGL4.47[luc2P/SIE/Hygro] Vector

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.

Available vectors and the pathways each can measure.

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.

Applications

• Transcription regulation.
• Virus-cell interactions.
• Compound screening.
• Post-translational modifications.
• GPCR signaling.
• Cell signaling.
• Promoter analysis.

For more information, see the Protocols & Applications Guide.