New generation of luciferase reporter gene vectors promises reduced background and lower risk of anomalous transcription
Promega has launched the new pGL4 luciferase reporter vectors which offer improved signal-to-background ratio and reduced risk of anomalous transcription.
This has been achieved by a dramatic reduction in the number of consensus transcription factor binding sites and regulatory elements on both the vector backbone and reporter genes.
A number of different configurations of vectors are available, each including either the synthetic firefly luciferase (luc2 (Photinus pyralis)) or Renilla luciferase (hRluc (Renilla reniformis)) genes.
These have been codon optimised for more efficient expression in mammalian cells.
Also included in the series are the Rapid Response reporter vectors.
These vectors give faster responses to changes in transcriptional activity with an increased relative magnitude of response, and are particularly well suited to monitor rapid processes such as promoter activation and repression.
The degradation sequences used in Rapid Response and pGL4 vectors are hPest and hCL1-Pest, both of which have been engineered for optimal expression in mammalian cells and reduced consensus transcription factor binding sites. Vector configurations include those without a promoter but including a multiple cloning site to allow a personal choice of promoter.
Promoter-containing pGL4 Luciferase reporter vectors utilise the SV40 early enhancer/promoter region, the HSV-TK promoter or the CMV immediate-early enhancer/promoter region.
All of the vector options also contain a SV40 late poly(A) signal 3' downstream of the reporter gene to increase the level of luciferase expression.
