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J. Biol. Chem. 273, 33885-33888. Cloning and characterization of the 5´-flanking region of the human growth hormone secretagogue receptor gene. 1998

Kaji, H., Tai, S., Okimura, Y., Iguchi, G., Takahashi, Y., Abe, H., Chihara, K.

Notes: Inserted promoter region and its deletion variants into pGL3-Basic Vector and co-transfected with pRL-CMV Vector (no ratio given). Measured luciferase activities with Dual-Luciferase® Reporter Assay System. (0952)

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J. Biol. Chem. 273, 33848-33855. Cloning and characterization of the human β4-integrin gene promoter and enhancers. 1998

Takaoka, A.S., Yamada, T., Gotoh, M., Kanai, Y., Imai, K., Hirohashi, S.

Notes: The authors performed DNaseI footprinting using the Core Footprinting System and nuclear extract or recombinant human AP-1 (c-jun). Promoter and enhancer elements of the human β4 integrin gene were analyzed using the Dual-Luciferase® Reporter Assay System. pRL-TK Vector was used as a transfection control. The transfection ratio of pGL3-Basic-derived plasmids to pRL-TK Vector was 10:1. (0296)

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J. Biol. Chem. 273, 7643-7649. Cloning of the multipartite promoter of the sodium-calcium exchanger gene NCX1 and characterization of its activity in vascular smooth muscle cells. 1998

Scheller, T., Kraev, A., Skinner, S., Carafoli, E.

Notes: Promoter activities were examined in primary rat vascular smooth muscle cells. The firefly luciferase vectors were co-transfected with the pRL-TK Vector at a 20:1 ratio. Transfections were performed with an unspecified member of the Tfx™ Reagent trio. (0451)

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J. Biol. Chem. 273, 26969-26976. Down-regulation of cytochrome P450 1A1 gene promoter by oxidative stress: Critical contribution of nuclear factor 1. 1998

Morel, Y., Barouki, R.

Notes: Promoter activities were studied in HepG2 cells with the Dual-Luciferase® Reporter Assay System. The Renilla control vector was constructed from the pRL-SV Vector by replacing the SV40 promoter with the α-globin gene proximal promoter (–79-+1). The Firefly luciferase construct was prepared in the pGL3 Basic Vector. The pGL3-based Vector and pRL-based Vector were transfected at a 3:1 ratio. A mutant nuclear factor 1 site was introduced into the 1A1 promoter with the GeneEditor™ in vitro Site-Directed Mutagenesis System by converting GCCA to CGCA. (0659)

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J. Biol. Chem. 273, 9457-9464. Genomic organization, chromosomal mapping, and promoter analysis of the mouse dentin sialophophoprotein (Dspp) gene, which codes for both dentin sialoprotein and dentin phosphoprotein. 1998

Feng, J.Q., Luan, X., Wallace, J., Jing, D., Ohshima, T., Kulkarni, A.B., D'Souza, R.N., Kozak, C.A., MacDougal, M.

Notes: Reporter assays were performed in the odontoblast cell line, MO6-G3. Experimental promoter constructs were assembled in the pGL3 Basic Vector and transfections were control through the use of the pRL-SV Vector. Luciferase activity was measured with the Dual-Luciferase® Reporter Assay System. Primer extension analysis of the gene transcript was performed with the Primer Extension System. (1196)

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J. Biol. Chem. 273, 25466-25471. Heat shock factor 1 mediates hemin-induced hsp70 gene transcription in K562 erythroleukemia cells 1998

Yoshima, T., Yura, T., Yanagi, H.

Notes: Reporter studies were performed in K562 cells. The Dual-Luciferase® Reporter Assay System was used to measure the firefly luciferase from a GAL4 binding site-containing luciferase vector and cotransformed pRL-SV40 Vector. No ratios were reported. Other vectors were transfected as well and produced proteins that activated that bound the GAL4 domain and activated transcription of the firefly luciferase in a two-hybrid-type assay. Another vector was constructed with Heat Shock Elements instead of GAL4 domains in a pGL3 Promoter Vector. (0116)

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J. Biol. Chem. 273, 33741. Identification of the cis-acting endoplasmic reticulum stress response element responsible for transcriptional induction of mammalian glucose-regulated proteins: Involvement of basic leucine zipper transcription factors. 1998

Yoshida, H., Haze, K., Yanagi, H., Yura, T., Mori, K.

Notes: Promoter studies were performed in HeLa cells. The promoter constructs were assembled in the pGL3 Basic Vector (10µg) and cotransformed with the pRL-SV40 Vector (0.1µg) at a 10:1 ratio using the calcium phosphate method. Reporter activity was measured with the Dual-Luciferase® Reporter Assay System. The TNT® T7 Quick Coupled Transcription/Translation System was used to produce a positive control protein for Western blot analysis of transfected HeLa cell extracts. (0115)

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Am. J. Physiol. 274, C681-C687. In vivo analysis of the myosin heavy chain IIB promoter region. 1998

Swoap, S. J.

Notes: The authors cloned a 2.6kb promoter-enhancer region of the MHC IIB gene into the pGL3-Basic Vector. They injected this construct along with the pRL-CMV Vector into mouse muscle cells. Tissue extracts were prepared with a buffered Tris solution containing a protease inhibitor cocktail and assayed for reporter activity using the Dual-Luciferase® Reporter Assay System. (0287)

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J. Biol. Chem. 273, 22120-22127. Induction of cyclooxygenase-2 by activated ha-ras oncogene in Rat-1 fibroblasts and the role of mitogen-activated protein kinase pathway. 1998

Sheng, H., Williams, C.S., Shao, J., Liang, P., DuBois, R.N., Beauchamp, R.D.

Notes: Luciferase studies were performed in a Rat-1 fibroblast cell line with an inducible Ha-ras gene. Experimental pGL3 Basic vectors were cotransfected with pRL-TK at a 2:1 ratio. (0389)

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Proc. Natl. Acad. Sci. USA 95, 8170-8174. Racial variability in the UDP-glucuronosyltransferase 1 (UGT1A1) promoter: A balanced polymorphism for regulation of bilirubin metabolism? 1998

Beutler, E., Gelbart, T. and Demina, A.

Notes: Studies were performed in HepG2 and HuH7 cells (both human cells of hepatic origin). One microgram of the experimental constructs in the pGL3 Basic Vector was cotransfected with 50ng of the pRL-SV Vector (20:1 ratio). Luciferase activity was measured using the Dual-Luciferase® Reporter Assay System. (1443)

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J. Biol. Chem. 273, 32312-32321. Repression of cyclooxygenase-2 and prostaglandin E2 release by dexamethasone occurs by transciptional and post-transcriptional mechanisms involving loss of polyadenylated mRNA 1998

Newton, R., Seybold, J., Kuitert, L.M.E., Bergmann, M. , Barnes, P.J.

Notes: A reporter construct was assembled in the pGL3 Control Vector downstream of the Xba I consisting of the 3'UTR of the COX-2 gene. The construct was designed to see if the UTR had an affect on mRNA stability in the presence of IL-1beta. The construct as well as the pSV-Beta Galactosidase Control Vector were transfected into A549 cells using the Tfx™-50 Reagent. A lot of detail is provided for the transfection. The reporter activities were monitored with the Luciferase Assay System and the Beta-Galactosidase Enzyme Assay System. (0606)

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J. Biol. Chem. 273, 34775. The roles of nuclear factor of activated T cells and Ying-Yang 1 in activation-induced expression of the interferon-gamma promoter in T cells. 1998

Sweetser, M.T., Hoey, T, Sun, Y.L., Weaver, W.M., Price, G.A., Wilson, C.B.

Notes: Reporter assays were performed in primary murine splenocytes. The interferon-gamma promoter constructs were assembled in the pGL3-Basic Vector and a Renilla luciferase control vector was constructed with the pRL-null Vector and a β-actin promoter. The plasmids were electroporated at a 10:1 ratio. Luciferase activities were followed with the Dual-Luciferase® Reporter Assay System. (0285)

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J. Lipid Res. 39, 1520-1524.. The –514 polymorphism in the hepatic lipase gene (LIPC) does not influence androgen-mediated stimulation of hepatic lipase activity. 1998

Vega, G.L., Gao, J., Bersot, T.P., Mahley, R.W., Verstraete, R., Grundy, S.M., White, A., Cohen, J.C.

Notes: Studies were performed in HepG2 cells. Experimental promoter contructs were assembled in the pGL3-Basic Vector and transfection efficiency was monitored by cotransfecting the pRL-CMV Vector. Luciferase activities were assayed with the Dual-Luciferase® Reporter Assay System. The ratio of experimental:control vector were not reported. (0219)

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Biochem. J. 332, 773-780. Transcript heterogeneity of the human reduced folate carrier results from the use of multiple promoters and variable splicing of alternative upstream exons. 1998

Zhang, L. , Wong, S. C. , Matherly, L. H.

Notes: The authors used the PolyATtract® mRNA Isolation System to isolate mRNA from total RNA. They used this mRNA-rich fraction for primer extension analysis using Promega's AMV Reverse Transcriptase. The Wizard® Plus Midiprep DNA Purification System was used for various plasmid isolations and the Erase-a-Base®  System was used to generate deletion series. The Dual-Luciferase® Reporter Assay System was used to study promoters cloned into the pGL3-Basic Vector. The pRL-SV40 was used as a transfection control plasmid. (0095)

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J. Biol. Chem. 273, 5607-5614. Transcriptional activation of the UDP-glucuronosyltransferase 1A7 gene in rat liver by aryl hydrocarbon receptor ligands and oltipraz. 1998

Metz, R.P. and Ritter, J.K

Notes: The RNasin Ribonuclease Inhibitor was used to protect RNA transcripts during primer extension analysis and in vitro transcription of RNA probes. The resultant RNA probes were used for RNase protection assays. (1639)

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J. Biol. Chem. 273, 14885-14890. Transcriptional regulation of endothelial nitric-oxide synthase by lysophosphatidylcholine 1998

Cieslik, K., Zembowicz, A., Tang, J.-L., Wu, K.K.

Notes: The Serine/Threonine Phosphatase Assay System was used to assess protein phosphatase 2A, 2B and 2C activity in nuclear extracts of Human umbilical vein cells (HUVEC) with or without lysophosphatidylcholine treatment. Gel shifts were also performed with nuclear extracts prepared from the HUVEC cells using the SP1 Consensus Oligonucleotide. Luciferase reporter studies were also performed in HUVEC cells using pGL3 Basic-derived vectors and the Luciferase Assay System. (1305)

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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. Immunol. 161, 3010-3018. Two distinct phospholipases C of Listeria monocytogenes induce ceramide generation, nuclear factor-kappaB activation, and E-selectin expression in human endothelial cells. 1998

Schwarzer, N., Nöst, R., Seybold, J., Parida, S.K., Fuhrmann, O., Krüll, M., Schmidt, R., Newton, R., Hippenstiel, S., Domann, E., Chakraborty, T., Suttorp, N.

Notes: A NF-kappaB luciferase reporter vector was constructed with the TATA box and transcription start site of the rabbit beta-globin promoter and three tandem repeats of the NF-kappaB consensus site in the pGL3 Basic Vector. Another vector was constructed with mutated NF-kappaB sites. The reporter vector was transfected into primary human umbilical vein endothelial cells with the Tfx™-50 Reagent. No details of the transfection conditions were provided. Luciferase activities were monitored with the Luciferase Assay System. (0418)

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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, 10551-10557.. A common pro-opiomelanocortin-binding element mediates leukemia inhibitory factor and corticotropin-releasing hormone transcriptional synergy. 1997

Bousquet, C. , Ray, D. W. , Melmed, S.

Notes: Luciferase studies were performed in AtT20 cells using the Luciferase Assay System. Experimental constructs were prepared in the pGL3 Basic Vector. (1392)

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J. Biol. Chem. 272, 21045-21051. A type I interferon signaling factor, ISF21, encoded on chromosome 21 is distinct from receptor components and their down-regulation and is necessary for transcriptional activation of interferon-regulated genes. 1997

Holland, K.A., Owczarek, C.M., Hwang, S.Y., Tymms, M.J., Constantinescu, S.N., Pfeffer, L.M., Kola, I., Hertzog, P.J.

Notes: Luciferase and CAT assays were conducted in CHO-K1 cells using constructs prepared in the pGL3-Basic and pCAT®-Basic Vectors, and the activity was normalized to β-galactosidase activity supplied by the pSV-β-Galactosidase Control Vector. The pCAT® Vectors have been replaced by the next-generation pCAT®3 Vectors. (1055)

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J. Biol. Chem. 272, 18316-18324. Binding of upstream stimulatory factor and a cell-specific activator to the calcitonin/calcitonin gene-related peptide enhancer. 1997

Lanigan, T.M., Russo, A.F.

Notes: Luciferase studies were performed in CA77 thyroid C cell line UI-0 using constructs prepared in the pGL3-Basic Vector. (0838)

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J. Biol. Chem. 272, 22199-22206. Butyrate activates the WAF1/Cip1 gene promoter through Sp1 sites in a p53-negative human colon cancer cell line. 1997

Nakano, K., Mizuno, T., Sowa, Y., Orita, T., Yoshino, T., Okuyama, Y., Fujita, T., Ohtani-Fujita, N., Matsukawa, Y., Tokino, T., Yamagishi, H., Oka, T., Nomura, H., Sakai, T.

Notes: Reporter studies were performed in WiDR human adenocarcinoma cells and MG63 human osteosarcoma cells. Experimental constructs were prepared in either the pGL2 Basic Vector or the pGL3 Basic Vector. (0644)

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J. Biol. Chem. 272, 16498-16506. Characterization and functional analysis of the promoter of RAGE, the receptor for advanced glycation end products 1997

Li, J., Schmidt, A.M.

Notes: pGL3 Basic Vector and pSV-β-galactosidase Control Vector were used in bovine aortic endothelial cells and rat vascular smooth muscle cells. The NF-κB p50 protein was used for DNase Footprinting (2-20 gel shift units). HeLaScribe® Nuclear Extract, Gel Shift Grade, was used as a positive control for NF-κB in a gel shift assay. (0793)

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Proc. Natl. Acad. Sci. USA 94, 9487-9492. Characterization of the α1B-adrenergic receptor gene promoter region and hypoxia regulatory elements in vascular smooth muscle. 1997

Eckhart, A.D. , Yang, N. , Xin, X. , Faber, J.E.

Notes: Luciferase studies were performed in aortic and vena caval smooth muscle cells. Transfections were normalized to β-galactosidase activity. Researchers report that pGL3 Promoter and pGL3 Control Vectors were 55-± 3-fold and 340±41-fold greater activity, respectively, than the pGL3 Basic Vector alone. The CREB consensus oligo was used in gel shifts of aortic and vena cava smooth muscle cell nuclear extracts. (1214)

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