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| 1. |
Wruck, C.J., Götz, M.E., Herdegen, T., Varoga, D., Brandenburg, L-O. and Pufe, T.
(2008)
Kavalactones protect neural cells against amyloid β peptide-induced neurotoxicity via ERK1/2-dependent Nrf2-activation
Molecular Pharmacology Fast Forward
March 11, 2008
,
epub ahead of print
.
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Notes:
The accumulation of the toxic form of the Amyloid-β peptide is known to induce oxidative damage in the brain. Although treatment with antioxidants has not proven effective at controlling AD symptoms, inducing the natural systems in the brain that protect from oxidative damage may provide a possible therapeutic approach. A host of antioxidant and detoxifying enzymes are upregulated by binding of the Nrf2 transcription factor to the ARE (antioxidant response element) regulatory sequence. The authors used a Dual Luciferase® Reporter Assay to assess modulation of gene activity through ARE by kavalactones. Kavalactones are compounds found in the roots and rhizomes of Kava (Piper methysticum), a plant cultivated an used in some Pacific societies for medicinal and social uses. The ARE1 region from the rat NAD(P)H:quinone oxidoreductase-1 gene was placed upstream of a pGL3 firefly luciferase reporter construct and cotransfected along with a pRL-TK Renilla control construct into PC12 or C6 cells. The data show induction of luciferase activity by kavalactones. Further investigation shows that the kavalactones promote Nrf2 stabilization possibly through the ERK1/2 pathway.
(0003859) |
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Products: Dual-Luciferase® Reporter Assay System | pGL3-Basic Vector | pGL3-Control Vector | pGL3-Enhancer Vector | pGL3-Promoter Vector | pRL-TK Vector |
| 2. |
Purbey, P.K., Singh, S., Kumar, P.P., Mehta, S., Ganesh, K.N., Mitra, D. and Galande, S.
(2008)
PDZ domain-mediated dimerization and homeodomain-directed specificity are required for high-affinity DNA binding by SATB1.
Nucleic Acids Res.
36
,
2107–2722
.
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Notes:
To learn about the ideal target binding sequence for SATB1, the T-lineage-enriched chromatin organizer and transcription factor, random oligonucleotides underwent SELEX and five rounds of selection by EMSA. The enriched library of oligos was cloned into the pGEM®-T Easy Vector, transformed and sequenced. Several variants of SATB1-binding consensus sequences were annealed, ligated into the pGL3-Promoter Vector and cotransfected into HEK 293 cells with a plasmid that either contained SATB1 or was empty. After 48 hours, the cells were harvested and luciferase activity measured. The CheckMate™ Mammalian Two-Hybrid System was used to assess how the N-terminal PDZ domain of SATB1 interacted with the Cut and homeodomain in the C-terminus.
(0003982) |
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Products: CheckMate™ Mammalian Two-Hybrid System | pGEM®-T Easy Vector System I | pGEM®-T Easy Vector System II | pGL3-Promoter Vector |
| 3. |
Zhang, X., Horrell, S.A., Delaney, D., Gottlieb, D.I.
(2008)
Embryonic Stem Cells as a Platform for Analyzing Neural Gene Transcription
Stem Cells
26
,
1841-1849
.
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Notes:
The authors note that while spatially and temporally specific gene transcription is a fundamental process in the normal development of mammalian stem cells, transcription in stem cells is currently studied by a set of methodologies with significant limitations. For instance, transient transfections analyze gene regulatory elements in nonchromosomal context. Using transgenic mice places transgenes in chromosomal context, however the chromosomal site where the transgene is inserted strongly influences the transgenes expression. As well, the need to make transgenic mice limits the number of experiments that can be done. ESCs can overcome these limitation. Undifferentiated stem cells are suitable for genetic engineering approaches such as gene targeting and recombinase-mediated cassette exchanges. By using such techniques, precisely planned alteration of native genes such as insertion of reporters, deletions of nearby or distant DNA sequences and mutational substitutions can be made. The authors wanted to analyze the Olig2 gene, a helix-loop-helix transcription factor expressed in the developing nervous system. Because Olig2 plays a central role in differentiation, understanding how it is regulated is important to understanding the larger transcriptional network controling development.
To this end, the authors used vectors for transient transfection experiments, constructed by amplifying regions of the Olig2 gene by PCR using primers tailed with appropriate restrictions sites and cloning the fragments into a pGL3 Luciferase Reporter Vector (Cat. # E1741, e1751, e1761, e1771). Promoter-reporter DNA was transfected into ESCs, cells were cultured 24 hours, then luciferase assays (Promega, type not specified) used to measure transgene expression.
(0003918) |
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| 4. |
Tan, K.P., Yang, M. and Ito, S.
(2007)
Activation of Nrf2 by toxic bile acids provokes adaptive defense responses to enhance cell survival at the emergence of oxidative stress.
Mol. Pharmacol.
72
,
1380–1390
.
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Notes:
The authors explored the role that nuclear factor (erythroid 2-like) factor 2 (Nrf2) may have in mitigating the cytotoxic effects of bile acids on cells. A reporter vector was constructed using the core sequence of antioxidant reporter element (ARE) sythnesized by annealing two complementary oligonucleotides with Kpn1 and BglII at the 5’ and 3’ ends, respectively, and ligated into the same restriction sites of the pGL3-Promoter Vector (designated pGL3_ARE). To ensure specificity for the experiments, three point mutations were introduced into the ARE sequence (designated pGL3_mARE). To create HepG2 cells that stably expressed human Na(+)-taurocholate co-transporting polypeptide (NTCP), the cDNA clone of NTCP was subcloned into pTargeT™ Mammalian Expression Vector via the NotI site and selected using 500μg/ml G-418. The stable clones or standard HepG2 cells were transiently transfected with 0.1µg of pGL3_ARE or pGL3_mARE, 0.02µg of the control reporter pRL-TK Vector, with or without Nrf2 or dominant negative Nrf2 expression constructs. After overnight transfection, the cells were treated with bile acids for 16–18 hours and luciferase activities determined using the Dual-Luciferase® Reporter Assay System. Each experiment was done in triplicate and repeated at least two times.
(0003691) |
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Products: Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | pGL3-Promoter Vector | pRL-TK Vector | pTARGET™ Mammalian Expression Vector System |
| 5. |
Zhang, Y-w., Wang, R., Liu, Q., Zhang, H., Liao, F-F. and Xu, H.
(2007)
Presenilin/γ-secretase-dependent processing of β-amyloid precursor protein regulates EGF receptor expression
Proc. Natl. Acad. Sci. U S A
104
,
10613-10618
.
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Notes:
The authors of this study investigated the downstream effects of the release of the intracellular domain of the Amyloid-β precursor protein (AICD) on cellular activities. They amplified the 5′ region of the mouse EGFR gene and cloned it into a pGL3 vector. This construct was cotransfected into embryonic fibroblasts derived from APP/APLP2 DKO mice along with a vector expressing AICD, AICD and the multidomain protein Fe65, Fe65 alone or NotchΔE, along with a Renilla control vector to normalize data for transfection efficiency. The data indicate that AICD negatively regulates transcription of the EGF Receptor gene.
(0003861) |
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Products: pGL3-Promoter Vector | pRL-SV40 Vector |
| 6. |
Kanayama, T., Arito, M., So, K., Hachimura, S., Inoue, J. and Sato, R.
(2007)
Interaction between sterol regulatory element-binding proteins and liver receptor homolog-1 reciprocally suppresses their transcriptional activities.
J. Biol. Chem.
282
,
10290–10298
.
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Notes:
To explore the interaction of liver receptor homolog (LRH)-1, a known suppressor of sterol regulatory element-binding protein (SREBP) transcriptional activity, human LRH-1 was reverse transcribed then amplified by PCR from total RNA from HepG2 cells. The amplification product was ligated into the pTargeT™ Mammalian Expression Vector to create pTarget-LRH1. For reporter experiments, a PCR fragment that encompassed the 1.3kb 5’-promoter region of the human small heterodimer partner (SHP) gene was cloned into the pGL3-Basic Vector (designated pSRB). The pGL3-Promoter Vector was used to construct pLRHREx3, which contains three LRH-1 response elements, and the insert was generated using synthetic oligonucleotides. HEK293 cells were cotransfected with 0.2µg of a promoter-firefly luciferase construct, 0.1µg of a SREBP expression plasmid, 10ng of phRL-TK Vector and 0.2 or 0.6µg of pTarget-LRH1. Alternatively, the cotransfected plasmids were 0.2µg of pSHP, 0.1µg of pTarget-LRH1, 10ng of phRL-TK Vector and 0.2 or 0.6µg of a SREBP expression plasmid. The pLRHREx3 construct (0.2µg) was cotransfected with 0.1µg of a LRH-1 expression plasmid, 0.2µg of pCMXPGC-1α (peroxisome proliferator activated receptor γ coactivator-1α), 10ng of phRL-TK Vector, and 0.1 or 0.3µg of a pSREBP expression vector in HEK 293 cells. Luciferase expression was assayed 48 hours post-transfection using the Dual-Luciferase® Assay Reporter System. To express SREBPs and LRH-1 in vitro, inserts were ligated into the pTNT™ Vector, synthesized using the TNT® Coupled Transcription/Translation System with radiolabeled methionine. Ten microliters of the 35S-labelled protein was then used in a GST-pulldown assay.
(0003692) |
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Products: Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | pGL3-Basic Vector | pGL3-Promoter Vector | pTNT™ Vector | TNT® SP6 Coupled Reticulocyte Lysate System | TNT® SP6 Coupled Reticulocyte Lysate System, Trial Size | TNT® T3 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System | TNT® T7 Coupled Reticulocyte Lysate System, Trial Size |
| 7. |
Tsuchiya, Y., Nakajima, M., Takagi, S., Taniya, T., and Yokoi, T.
(2006)
MicroRNA regulates the expression of human cytochrome P450 1B1.
Cancer Res.
66
,
9090-9098
.
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Notes:
These authors identified a region complementary to the microRNA miR-27b in the 3´ UTR of the cytochrome p450 CYP1B1 mRNA, and showed that miR-27b was involved in regulation of CYP1B1 expression. The 3´ UTR containing the miRNA target site was cloned downstream of the luciferase gene in the pGL3 Promoter Vector and cotransfected into the miR-27b-positive breast cancer cell line MCF-7 and into miR-27b-negative Jurkat cells. Luciferase expression levels from the reporter vector containing the CYP1B1 3´ UTR sequence were reduced in miR-27b-positive cells, but not in the Jurkat cell controls. Delivery of an antisense oligoribonucleotide directed against miR-27b to MCF-7 cells containing the reporter construct resulted in restoration of luciferase activity. The effects of inhibition of miR-27b on protein levels and enzymatic activity of CYP1B1 were then investigated in MCF-7 cells. CYP1B1 protein levels and enzymatic activity increased significantly in cells transfected with the antisense oligo; the enzymatic activity was measured using a p450-Glo™ Assay. The coding region and 3´ UTR of the CYP1B1 gene were also PCR-amplified, subcloned the into the pTargeT™ Mammalian Expression Vector, and transfected into HEK293 cells. The effect of overexpression of miR-27b on protein levels and enzymatic activity of CYP1B1 was then evaluated in these cells.
(0003622) |
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Products: Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | P450-Glo™ CYP1B1 Assay | pGL3-Promoter Vector | pTARGET™ Mammalian Expression Vector System |
| 8. |
Guindalini, C., Howard, M., Haddley, K., Laranjeira, R., Collier, D., Ammar, N., Craig, I., O'Gara, C., Bubb, V.J., Greenwood, T., Kelsoe, J., Asherson, P., Murray, R.M., Castelo, A., Quinn, J.P., Vallada, H., and Breen, G.
(2006)
A dopamine transporter gene functional variant associated with cocaine abuse in a Brazilian sample.
Proc. Natl. Acad. Sci. U S A
103
,
4552-4557
.
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Notes:
These authors investigated the effect of various polymorphisms in the dopamine transporter gene (SLC6A3) on susceptibility to cocaine addiction. Genotyping of various polymorphisms in cocaine abusers and control subjects revealed a potential association of the int8 VNTR with cocaine abuse. Seven alleles of the int8 VNTR were sequenced. Various allelic sequences were then cloned into a modified phRL-SV40 Renilla luciferase reporter vector and transfected into the mouse SN4741 cell line, which expresses the dopamine transporter, and the effects on reporter activity were monitored. Sequences of two alleles were then cloned into a pGL3 Promoter Vector construct and transfected into JAP cells. The cells were then challenged with various amounts of cocaine, KCL or KCl and forskolin, and the effect on reporter activity was monitored. The TransFast™ Reagent was used for transfections at a 2:1 reagent:DNA ratio.
(0003543) |
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Products: pGL3-Promoter Vector | TransFast™ Transfection Reagent |
| 9. |
Koon, H-W., Zhao, D., Zhan, Y., Rhee, S.H., Moyer, M.P. and Pothoulakis, C.
(2006)
Substance P stimulates cyclooxygenase-2 and prostaglandin E2 expression through JAK-STAT activation in human colonic epithelial cells.
J. Immunol.
176
,
5050-9
.
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Notes:
The COX-2 promoter was cloned by PCR into the pGL3 Vector. NCM460-NK-1R cells were transiently transfected with the cloned promoter and the pRL-TK vector as an internal control or siRNA targeted against various JAK/STAT genes. The Dual-Luciferase® Assay was used to measure promoter activity. The wildtype COX-2 promoter was mutagenized using the GeneEditor™ in vitro Site-Directed Mutagenesis System.
(0003384) |
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Products: Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | GeneEditor™ in vitro Site-Directed Mutagenesis System | pGL3-Basic Vector | pGL3-Control Vector | pGL3-Enhancer Vector | pGL3-Promoter Vector |
| 10. |
Chew, J.L., Loh, Y.H., Zhang, W., Chen, X., Tam, W.L., Yeap, L.S., Li, P., Ang, Y.S., Lim, B., Robson, P. and Ng, H.H.
(2005)
Reciprocal transcriptional regulation of Pou5f1 and Sox2 via the Oct4/Sox2 complex in embryonic stem cells.
Mol. Cell. Biol.
25
,
6031–46
.
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Notes:
The authors studied the effects of Embryonic Stem Cell (ESC)-specific regulation on the Pou5f1 promoter in human and mouse cells. To examine the effect of knockdown of Oct4 and Sox2 (two genes involved in ESC regulation) on the Pou5f1 promoter, a 3kb fragment of the human POU5F1 promoter was cloned into pGL3-Basic Vector and 100ng cotransfected with 100ng shRNA plasmids into mouse E14 ESCs. Five nanograms of pRL-SV40 Vector served as a transfection control. For the enhancer assay, a 461bp fragment of genomic DNA containing the SRR2 enhancer of Sox2 was amplified and cloned into the pGL3-Promoter Vector. The same amounts of plasmid, shRNA and transfection control were transfected into E14 ESCs as in the Pou5f1 promoter assay. To investigate gene knockdown in 293T cells, 5ng of the two open reading frame (ORF) constructs (the Luc-Sox2 and the Luc-Pou5f1 ORFs cloned into the psiCHECK™-2 Vector) were cotransfected with 100ng shRNA plasmid. The outcome was examined 48–60 hours post-transfection using the Dual-Luciferase® Reporter Assay System.
(0003291) |
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Products: Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | pGL3-Basic Vector | pGL3-Promoter Vector | pRL-SV40 Vector | psiCHECK™-2 Vector |
| 11. |
Kurata, S., Okuyama, T., Osada, M., Watanabe, T., Tomimori, Y., Sato, S., Iwai, A., Tsuji, T., Ikawa, Y. and Katoh, I.
(2004)
p51/p63 controls subunit 3 of the major epidermis integrin anchoring the stem cells to the niche.
J. Cell Biol.
279
,
50069–50077
.
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Notes:
HeLa and Saos-2 were transiently transfected with luciferase reporter vectors made from the pGL3-Promoter vector containing various intron sequences from the human integrinα3 gene ITGA3. The first intron sequence was originally cloned by PCR cloning into the pGEM®-T Easy vector. Cell cultures were assessed for luciferase activity by making lysates with the Glo Lysis Buffer and assaying with the Steady-Glo® Assay kit.
(0003225) |
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Products: Glo Lysis Buffer, 1X | pGEM®-T Easy Vector System I | pGEM®-T Easy Vector System II | pGL3-Promoter Vector | Steady-Glo® Luciferase Assay System |
| 12. |
Wu, X., Fan, W., Xu, S., and Zhou, Y.
(2003)
Sensitization to the cytotoxicity of cisplatin by transfection with nucleotide excision repair gene Xeroderma Pigmentosun Group A antisense RNA in human lung adenocarcinoma cells.
Clin. Cancer Res.
9
,
5874–5879
.
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Notes:
The pGL3-Promoter Vector was treated with various concentrations of the chemotherapeutic cancer and DNA cross-linking agent, cisplatin. The damaged pGL3-Promoter Vector and the pSV-β-Galactosidase Control Vector were then transiently transfected into the human lung adenocarcinoma cell line, A549, and later analyzed for luciferase activity using the Luciferase Assay System with Reporter Lysis Buffer. The researchers also used M-MLV Reverse Transcriptase to clone the antisense sequence to XPA (Xeroderma Pigmentosum group A) mRNA. The cloned antisense sequence was then expressed in A549 cells for its effect on XPA gene expression.
(0002833) |
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Products: Luciferase 1000 Assay System | Luciferase Assay System | Luciferase Assay System Freezer Pack | Luciferase Assay System with Reporter Lysis Buffer | Luciferase Assay System, 10-Pack | M-MLV Reverse Transcriptase | pGL3-Promoter Vector | pSV-β-Galactosidase Control Vector | Reporter Lysis 5X Buffer |
| 13. |
Mian, B.M., Dinney, C.P.N., Bermejo, C.E., Sweeny, P., Tellez, C., Yang, X.D., Gudas, J.M., McConkey, D.J., Bar-Eli, M.
(2003)
Fully human anti-interleukin 8 antibody inhibits tumor growth in orthotopic bladder cancer xenografts via down-regulation of matrix metalloproteases and nuclear factor-κB
Clin. Cancer Res.
9
,
3167-3175
.
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Notes:
Luciferase reporter gene constructs based on the pGL3 vector that contained either MMP-2 or MMP-9 promoters, SV40 promoter (positive control ) or the luciferase basic vectors were transfected into metastatic cells, S53JB-V cells or UMUC-3 cells along with a pβ-actin RL control construct. Cells were treated with an anti-IL-8 antibody, IgG control or no antibody. Dual luciferase reporter assays were performed to determine the effect of the treatments on activity and expression of the MMP-2 and MMP-9 constructs.
(0002716) |
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Products: Dual-Luciferase® Reporter Assay System | pGL3-Basic Vector | pGL3-Control Vector | pGL3-Enhancer Vector | pGL3-Promoter Vector |
| 14. |
Niimi, T., Hayashi, Y. and Sekiguchi, K.
(2003)
Identification of an upstream enhancer in the mouse laminin α1 gene defining its high level of expression in parietal endoderm cells.
J. Biol. Chem.
278
,
9332-9338
.
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Notes:
The pGL3-basic and pGL3-Promoter vectors were used to make constructs with varying lengths of the mouse LAMA1 promoter. Gene fragments up to 6.1 kb were cloned into the pGL3-promoter vector and analyzed for luciferase expression in a variety of cell lines using the Dual-Luciferase® Reporter Assay System. F9, NIH/3T3, PYS-2 and EHS cells were transiently transfected with 200ng of reporter construct and 20ng of the Renilla luciferase-expressing phRL-null vector in 24-well plates. Forty-eight hours later, cell lysates were harvested with Passive Lysis Buffer. Site-directed mutagenesis of the LAMA1 gene promoter was performed with the GeneEditor™ in vitro Site-Directed Mutagenesis System.
(0002631) |
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Products: Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | GeneEditor™ in vitro Site-Directed Mutagenesis System | pGL3-Basic Vector | pGL3-Control Vector | pGL3-Promoter Vector |
| 15. |
Zhu, X. and Craft, C.M.
(2000)
Modulation of CRX transactivation activity by phosducin isoforms
Mol. Cell. Biol.
20(14)
,
5216-5226
.
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Notes:
The pRL-null Vector was used as a reporter for a DLR® assay (Dual-Luciferase® Reporter Assay System). The promoter from human IRBP (interphotoreceptor retinoid binding protein) was cloned into the HindIII site of pRL-null. Transient transfections were done with 1µg of the pIRBP, 0.4µg of expression construct, and 0.2µg of pGL3-promoter as a transfection efficiency control. Activity of the Renilla luciferase was normalized to the activity of the firefly luciferase. Cells used for this assay were COS-7 and Weri-Rb-1 retinoblastoma cells.
(0002149) |
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Products: Dual-Luciferase® Reporter Assay System | pGL3-Promoter Vector | pRL-null Vector |
| 16. |
Stoneley, M., Paulin, F. E., Le Quesne, J. P., Chappell, S. A., Willis, A. E.
(1998)
C-Myc 5' untranslated region contains an internal ribosome entry segment.
Oncogene
16
,
423-428
.
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Notes:
The authors used the Dual-Luciferase® Reporter Assay System, pGL3 Control Vector, pRL-CMV Vector, pSV-beta-Galactosidase Control Vector, Luciferase Assay System and RNase ONE® Ribonuclease in their studies. The objective of the paper was to demonstrate that the 5' untranslated region (UTR) of c-myc functions as an internal ribosome entry site (IRES). To directly assay the ability of the UTR to function as a IRES, the Renilla luciferase gene was subcloned in front of the UTR. Dual expression of the luciferases was demonstrated in both HeLa cells and HepG2 cells and dependent upon the c-myc UTR. Mutation of the Renilla luciferase sequence to contain a hairpin structure, demonstrate that the Renilla luciferase activity could reduced by 75% with no effect on the firefly luciferase activity. RNase protection assays with a 624 nucleotide probe that crossed the Renilla luciferase cDNA, the c-myc UTR and the firefly luciferase cDNA demonstrated that only one single message is produced by the transfected HeLa cells and the c-myc UTR is not functioning as a promoter. All transfections were normalized to control β-galactosidase activity.
(0000339) |
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Products: Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Luciferase Assay System | pGL3-Control Vector | pGL3-Enhancer Vector | pGL3-Promoter Vector | pSV-β-Galactosidase Control Vector | RNase ONE™ Ribonuclease |
| 17. |
Yoshima, T., Yura, T., Yanagi, H.
(1998)
Heat shock factor 1 mediates hemin-induced hsp70 gene transcription in K562 erythroleukemia cells
J. Biol. Chem.
273
,
25466-25471
.
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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.
(0000116) |
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Products: Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | pGL3-Promoter Vector | pRL-SV40 Vector |
| 18. |
Sheng, H., Williams, C.S., Shao, J., Liang, P., DuBois, R.N., Beauchamp, R.D.
(1998)
Induction of cyclooxygenase-2 by activated ha-ras oncogene in Rat-1 fibroblasts and the role of mitogen-activated protein kinase pathway.
J. Biol. Chem.
273
,
22120-22127
.
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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.
(0000389) |
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Products: Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | pGL3-Basic Vector | pGL3-Promoter Vector | pRL-TK Vector |
| 19. |
Cerutti, A., Schaffer, A., Shah, S., Zan, H., Liou, H.-C., Goodwin, R.G., Casali, P.
(1998)
CD30 is a CD40-inducible molecule that negatively regulates CD40-mediated immunoglobulin class switching in non-antigen-selected human B cells
Immunity
9
,
247-256
.
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| |
Notes:
Luciferase studies were performed in the human B cell line, CL-01. Cells were electroporated with a 40µl DNA-TE buffer solution containing 20µg of constructs in either pGL3 Basic or pGL3 Promoter Vectors and 10ng of the pRL-CMV Vector. Thus, a 1:2000 ratio of firefly to Renilla luciferase vector is acheived and activity was measured with the Dual-Luciferase® Reporter Assay System.
(0001353) |
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Products: Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | pGL3-Basic Vector | pGL3-Promoter Vector | pRL-CMV Vector |
| 20. |
Eckhart, A.D. , Yang, N. , Xin, X. , Faber, J.E.
(1997)
Characterization of the α1B-adrenergic receptor gene promoter region and hypoxia regulatory elements in vascular smooth muscle.
Proc. Natl. Acad. Sci. U S A
94
,
9487-9492
.
|
| |
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.
(0001214) |
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Products: CREB Consensus Oligonucleotide | Luciferase Assay System | Luciferase Assay System with Reporter Lysis Buffer | pGL3-Basic Vector | pGL3-Control Vector | pGL3-Promoter Vector | pSV-β-Galactosidase Control Vector |
| 21. |
Coso, O. A. , Montaner, S. , Fromm, C. , Lacal, J. C. , Prywes, R. , Teramoto, H. , Gutkind, J. S.
(1997)
Signaling from G protein-coupled receptors to the c-jun promoter involves the MEF2 transcription factor. Evidence for a novel c-jun amino-terminal kinase-independent pathway
J. Biol. Chem.
272
,
20691-20697.
.
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| |
Notes:
Luciferase studies were performed in NIH3T3 cells expressing 20,000 muscarinic acetylcholine receptors per cell using constructs prepared in the pGL3 Promoter Vector.
(0001276) |
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Products: Luciferase Assay System with Reporter Lysis Buffer | pGL3-Promoter Vector |
| 22. |
Crossin, K.L., Tai, M.H., Krushel, L.A., Mauro, V.P. and Edelman, G.M.
(1997)
Glucocorticoid receptor pathways are involved in the inhibition of astrocyte proliferation.
Proc. Natl. Acad. Sci. U S A
94
,
2687-2692
.
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| |
Notes:
Studies performed in primary astrocytes.
(0001613) |
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Products: pGL3-Promoter Vector |
| 23. |
Xie, M. , Shao, G. , Buyse, I. M. , Huang, S.
(1997)
Transcriptional repression mediated by the PR domain zinc finger gene RIZ
J. Biol. Chem.
272
,
26360-26366
.
|
| |
Notes:
Expression of luciferase reporter constructs, derived from the pGL3 Promoter Vector, was studied in 3T3 cells.
(0000165) |
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Products: pGL3-Control Vector | pGL3-Promoter Vector |
| 24. |
Ko, B.C.B., Ruepp, B., Bohren, K.M., Gabbay, K.H., Chung, S.S.M.
(1997)
Identification and characterization of multiple osmotic response sequences in the human aldose reductase gene.
J. Biol. Chem.
272
,
16431-16437
.
|
| |
Notes:
Chang liver cells were transfected with constructs in pGL3-Basic and pGL3-Promoter Vectors using the Tfx™-50 Reagent. Constructs were verified with the fmol® DNA Cycle Sequencing System.
(0000911) |
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Products: fmol® DNA Cycle Sequencing System | pGL3-Basic Vector | pGL3-Control Vector | pGL3-Promoter Vector | Tfx™-50 Reagent |
| 25. |
Xie, Z. , Bikle, D. D.
(1997)
Cloning of the human phospholipase C-gamma1 promoter and identification of a DR6-type vitamin D-responsive element.
J. Biol. Chem.
272
,
6573-6577
.
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Notes:
Expression of luciferase reporter constructs, derived from the pGL3 Vectors, was studied in human keratinocytes.
(0000166) |
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Products: pGL3-Basic Vector | pGL3-Control Vector | pGL3-Promoter Vector |