The pGL3 Luciferase Reporter Vectors provide a basis for the quantitative analysis of factors that potentially regulate mammalian gene expression. These may be cis- or trans-acting factors. The backbone of the pGL2 Luciferase Reporter Vectors was redesigned for the pGL3 Vectors for increased expression, with a modified coding region for firefly (Photinus pyralis) luciferase that has been optimized for monitoring transcriptional activity in transfected eukaryotic cells. The assay of this genetic reporter is rapid, sensitive and quantitative. In addition, the Luciferase Reporter Vectors contain numerous features aiding in the structural characterization of the putative regulatory sequences under investigation.
For the most advanced reporter vectors and widest selection of features, please see the pGL4 Luciferase Reporter Vectors.
The specific transcriptional characteristics of the pGL3 Vectors will vary for different cell types. This may be particularly true for COS cells, which contain the SV40 large T antigen that promotes replication from the SV40 origin found in the promoter of the pGL3-Promoter and pGL3-Control Vectors. The combination of the large T antigen and the SV40 origin will result in a higher copy number of these vectors in COS cells, which in turn may result in increased expression of the reporter gene compared to other cell and vector combinations.
The pGL3 Luciferase Reporter Vectors offer different combinations of the SV40 early promoter and enhancer sequences to provide flexible experimental strategies and user convenience. Functional mapping of genetic elements can be achieved through nested deletions, and single-stranded DNA may be prepared for mutagenesis or sequencing. Primers are available for sequencing across cloning sites and for primer extension analysis.
About the Vector Options
The reporter vectors are available in four options, depending on your needs. The Basic Vector has the reporter gene and restriction enzymes sites to clone in a promoter of interest. The Enhancer Vector contains the SV40 Enhancer Region for putative promoters that may need an additional regulator region. The Promoter Vector is suitable for cloning and testing putative enhancer regions. The Control Vector contains the SV40 Promoter and Enhancer Region and can be used as a normalization control for expression experiments.
pGL3 Luciferase Reporter Vectors Technical Manual
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