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J. Biol. Chem. 273, 11923-11929. Transcriptional regulation of the prothrombin gene in muscle. 1998

Kim, S., Nelson, P.G.

Notes: Studies were performed in HepG2 cells and mouse C2 skeletal muscle cells. The experimental pGL3-Basic Vector-derived constructs were cotransfected with pRL-CMV Vector using the calcium phosphate method. Luciferase activity was monitored with the Dual-Luciferase® Reporter Assay System 48 hours after transfection. (0899)

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J. Biol. Chem. 273, 29816-29821. Two YY-1-binding proximal elements regulate the promoter strength of the TATA-less mouse ribonucleotide reductase R1 gene. 1998

Johansson, E., Hjortsberg, K., Thelander, L.

Notes: The authors cloned promoter region (and deletion mutants) of mouse ribonucleotide reductase R1 gene into pGL3-Basic Vector. Also, they cloned 3´-UTR of this gene downstream of firefly luciferase coding region in pGL3-Control Vector to study posttranscriptional effects mediated by this region. Co-transfected plasmids with pRL-SV40 into Balb/3T3 fibroblasts. Assayed luciferase activity with Dual-Luciferase® Reporter Assay System. (0985)

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J. Biol. Chem. 272, 31793-31800. CCAAT/Enhancer-binding protein delta activates insulin-like growth factor-I gene transcription in osteoblasts. Identification Of a novel cyclic amp signaling pathway in bone 1997

Umayahara, Y., Ji, C., Centrella, M., Rotwein, P., McCarthy, T.L.

Notes: Studies were performed in COS-7 cells. The experimental firefly luciferase vector (derived from pGL2-Basic Vector) was co-transfected with the Renilla control Vector (pRL-CMV Vector) and plasmids expressing either the CCAAT/Enhancer-binding protein beta or delta from a CMV promoter. The firefly luciferase:expression plasmid ratio was 10:1 and the firefly luciferase:Renilla luciferase vector ratio was 1000:1. In vitro translations were performed with the TNT® Coupled Reticulocyte Lysate System. (0252)

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J. Biol. Chem. 272, 31278-31284. Differential regulation of the transcriptional activity of the orphan nuclear receptor NGFI-B by membrane depolarization and nerve growth factor. 1997

Katagiri, Y., Hirata, Y., Milbrandt, J., Guroff, G.

Notes: Dual-Luciferase® Reporter Assay System was used to perform studies in rat PC12 cells. The firefly luciferase reporter vector was transfected with the control plasmid (pRL-TK Vector) at a 20:1 ratio. Eighteen hours after transfection, the cells were treated with NGF, EGF or KCl and assayed up to eight hours later. In some experiments, PC12 cells were co-transfected with the firefly luciferase plasmid, the Renilla control vector and a vector expressing the wildtype or mutant NGFI-B protein. (0920)

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Proc. Natl. Acad. Sci. USA 94, 12309-12313. Expression of hepatocyte nuclear factor 6 in rat liver is sex-dependent and regulated by growth hormone. 1997

Lahuna, O., Fernandez, L., Karlsson, H., Maiter, D., Lemaigre, F.P., Rousseau, G.G., Gustafsson, J., Mode, A.

Notes: Dual-Luciferase® Reporter Assay System studies were performed in HepG2 cells. The experimental plasmid constructed in pGL2-Basic Vector was used at a 40:1 ratio over the pRL-CMV Vector. The experimental and control plasmids were cotransfected with a third plasmid expressing hepatocyte nuclear factor 6 from a CMV promoter. The luciferase activity expressed in the presence or absence of the hepatocyte factor were measured. The TNT® Coupled Wheat Germ Extract System was used to in vitro translate four transcription factors for assay of gel shift with the promoter construct. (0870)

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J. Biol. Chem. 272, 30583-30588. Identification of a Placental Enhancer for the Human Leptin Gene 1997

Bi, S., Gavrilova, O., Gong, D.W., Mason, M.M. and Reitman, M.

Notes: Promoter activity was assayed in JEG-3 choriocarcinoma cells, JAR choriocarcinoma cells, HeLa cells and primary adipocytes. The pGL3 Vectors were transfected at a 20:1 ratio over pRL-CMV Vector. Enhancer activity was functional only in the choriocarcinoma cells. The Dual-Luciferase® Reporter Assay System was used to measure luciferase expression. (1446)

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Virology 238, 432-443. Increasing the ratio of PP2A core enzyme to holoenzyme inhibits Tat-stimulated HIV-1 transcription and virus production. 1997

Ruediger, R., Brewis, N., Ohst, K. and Walter, G.

Notes: This paper describes a method for quantitation of luciferase mRNA by in vitro translation using Promega TNT® Coupled Wheat Germ Extract System. (Rabbit reticulocyte lysate could not be used because of luciferase quenching problems). In this reporter gene assay, COS cells were transfected with a firefly luciferase reporter plasmid driven by promoters/enhancers of varying strengths and total cellular RNA was isolated and translated in vitro using the TNT® System. To normalize expression, a CMV Renilla luciferase or beta-galactosidase vector was used as a control. To measure luciferase activity either Promega Luciferase Assay System or Dual-Luciferase® Reporter Assay System was used. (2047)

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J. Biol. Chem. 272, 26620-26626. Regulation of clusterin gene expression by transforming growth factor β. 1997

Jin, G., Howe, P.H.

Notes: Luciferase reporter studies performed in CCL64 (Mv1Lu) mink lung cells using constructs prepared in the pGL2-Basic Vector and were normalized to β-galactosidase activity. Studies performed in 10T1/2, 3Tp, HeLa cells and primary bovine aortic endothelial cells were normalized to the Renilla luciferase. A five to one (w/w) ratio of experimental to control vector was used with pRL-CMV Control Vector. A two to one (w/w) ratio of experimental to control vector was used with pSV-β-Galactosidase Control Vector. (0981)

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J. Biol. Chem. 272, 31755-31763. Signal transduction-mediated activation of the aryl hydrocarbon receptor in rat hepatoma H4IIE cells. 1997

Backlund, M., Johansson, I., Mkrtchian, S., Ingelman-Sundberg, M.

Notes: Studies were performed in H4IIE rat hepatoma cells. The pGL3-based Vectors were co-transfected with the Renilla control vector (pRL-SV40 Vector) at a 6:1 ratio and assayed using the Dual-Luciferase® Reporter Assay System. (1489)

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J. Biol. Chem. 272, 17112-17117. Structure of the m1 muscarinic acetylcholine receptor gene and its promoter 1997

Pepitoni, S., Wood, I.C. Buckley, N.J.

Notes: The Dual-Luciferase® Reporter Assay System was used to study transfections of IMR32 and NIH3T3 cells with firefly luciferase vector (pGL3 Basic) constructs. Transfections were controlled with co-transfected pRL-CMV Vector. Tth DNA Polymerase was used as a high temperature reverse transcriptase. SP6 RNA Polymerase was used to produce probes for RNase protection assays. (0558)

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J. Biol. Chem. 272, 30662-30671. The Human CC Chemokine Receptor 5 (CCR5) Gene. Multiple transcripts with 5'-end heterogeneity, dual promoter usage, and evidence for polymorphisms within the regulatory regions and noncoding exons 1997

Mummidi, S., Ahuja, S.S., McDaniel, B.L., Ahuja, S.K.

Notes: Reporter studies were performed in THP-1 monocytic cells, Jurkat T-cell leukemic cells and K562 myelogenous leukemia cells. Experimental promoter constructs were prepared in the pGL3 Basic Vector. Transfections were control by cotransfection with the pRL-CMV Vector, a source of Renilla luciferase. The activity of the two luciferases was determined with the Dual-Luciferase® Reporter Assay System. Ratios of pGL3-based Vectors to the pRL-CMV Vector were not reported but 0.5µg of the pRL-CMV Vector was used in each transfection. (0630)

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Genomics 46, 459-65. The structural organization of the human Na+/myo-inositol cotransporter (SLC5A3) gene and characterization of the promoter. 1997

Mallee, J.J., Atta, M.G., Lorica, V., Rim, J.S., Kwon, H.M., Lucente, A.D., Wang, Y., Berry, G.T.

Notes: Cloned promoter from human SLC5A3 gene upstream of firefly luciferase in pGL2 Basic Vector. As a positive control, the beta actin promoter was cloned into pGL2 Basic Vector. Co-transfected SLC5A3 promoter constructs with the beta actin construct and pRL-CMV Vector at a ratio of 1:40:1. Measured luciferase activities with the Dual-Luciferase® Reporter Assay System. (0726)

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J. Biol. Chem. 272, 23489-23497. Transcriptional regulation of the mouse presenilin-1 gene. 1997

Mitsuda, N., Roses, A.D. and Vitek, M.P.

Notes: The Dual-Luciferase™ Reporter System was used to quantitate the presenilin promoter activity in Neuro2a neuroblastoma cells, mouse P19 embryonal carcinoma cells and NIH 3T3 cells. Studies were also performed in P19 cells treated with retinoic acid to acquire a neuron-like phenotype and P19 cells treated with dimethyl sulfoxide to acquire a muscle-like phenotype. The presenilin promoter functioned best in the Neuro2a and neuron-like P19 cells. 5´-RACE products from mouse brain RNA were purified with the Wizard® PCR Preps System and cloned into the pGEM-T Vector . The cloned amplimers were sequenced and used as a template for amplification to produce truncation mutants to assess promoter activity. (1590)

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EMBO J. 16, 1080-1092. Tumor necrosis factor alpha-induced activation of c-jun N-terminal kinase is mediated by TRAF2. 1997

Reinhard, C., Shamoon, B., Shyamala, V., Williams, L.T.

Notes: A firefly luciferase reporter vector containing three copies of the Ig kappa NF-kB binding site was cotransfected with vectors expressing either TRAF2 with or without MAPK Kinase 4 or mutants of TRAF2. All firefly luciferase activity was normalized to Renilla luciferase activity provided by the pRL-TK Vector. These studies were performed in COS 7 cells. Luciferase activities were determined with the Dual-Luciferase® Reporter Assay System. (0508)

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J. Biol. Chem. 272, 30208-30214. Two naturally occurring insulin receptor tyrosine kinase domain mutants provide evidence that phosphoinositide 3-kinase activation alone is not sufficient for the mediation of insulin's metabolic and mitogenic effects. 1997

Krook, A., Whitehead, J.P., Dobson, S.P., Griffiths, M.R., Ouwens, M., Baker, C., Hayward, A.C., Sen, S.K., Maassen, J.A., Siddle, K., Tavaré, J.M., O'Rahilly, S.

Notes: CHO-K1 cells were transfected at ~80% confluency with four plasmids: (1) a CMV-driven vector expressing the wildtype or mutant human insulin receptor; (2) a pGL3-Basic-derived Vector containing five GAL4 binding sites upstream of the firefly luciferase reporter; (3) the pRL-CMV Control Vector; and (4) a fusion of the GAL4-binding domain and Elk-1 transcription factor. The Dual-Luciferase® Reporter Assay System was used to confirm that specific mutations in the insulin receptor negate its ability to activate the Elk-1 transcription factor. (0896)

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J. Biol. Chem. 272, 17097-17103. Vasoactive peptides modulate vascular endothelial cell growth factor production and endothelial cell proliferation and invasion 1997

Pedram, A., Razandi, M., Hu, R. M., Levin, E. R.

Notes: Vascular smooth muscle cells were transfected with a firefly luciferase reporter vector and pRL-SV40 as a control. Transfection efficiency was normalized to the Renilla luciferase activity as determined by the Dual-Luciferase® Reporter Assay System. (0555)

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