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Table 1 Reporter plasmids used in this study

From: Application of the dual-luciferase reporter assay to the analysis of promoter activity in Zebrafish embryos

Plasmid name Purpose Relevant features
pEGFPLuc Positive control for assaying the effectiveness of transfection Kanamycin/neomycin marker. This reporter plasmid encodes a fusion of eGFP and luciferase from the firefly Photinus pyralis driven by the human CMV immediate early promoter (Clontech, Cat.# 6169-1).
plessEGFPLuc Negative control Kanamycin/neomycin marker. This vector was created by removing the CMV promoter of pEGFPLuc with the restriction enzymes AseI and NheI, followed by blunting of 5'-cohesive ends and autoligation.
zfpTERT(1/3 Kb)-EGFPLuc Determination of the zfTERT promoter activity Kanamycin marker. This vector was created by replacing the CMV promoter of pEGFPLuc with a 1 or 3 Kb-fragment of the zebrafish telomerase promoter region.
pNF-κB::Luc Assessment of NF-κB activation Ampicillin marker. This vector carries a the cDNA encoding the firefly (P. pyralis) luciferase gene placed under the control of three synthetic copies of the κB consensus of the immunoglobulin κ-chain promoter cloned in the BamHI site located upstream of the conalbumin transcription start site [19].
pRL-CMV, pRL-TK and pRL-SV40 Normalization Ampicillin marker. The pRL vectors contain the cDNA encoding Renilla luciferase cloned from the anthozoan coelenterate Renilla reniformis (sea pansy). Three different promoter configurations are available; CMV, TK and SV40.
pRL-EF1α Normalization Ampicillin marker. This vector was obtained by inserting the EF1α promoter in the pRL-null vector (Promega, Cat. # E2271).
  1. Note: pEGFPLuc has been discontinued and is no longer available from Clontech; it can be obtained from the authors on request. Full sequences and maps can be found on the Clontech and Promega websites.