p5xNF-κB-Luc信号通路报告载体荧光素酶基因报告质粒cis-Reporter plasmid-BioVector NTCC质粒载体菌种细胞基因保藏中心

p5xNF-κB-Luc

p5xNF-κB-Luc信号通路报告载体荧光素酶基因报告质粒cis-Reporter plasmid，含有NF-κB（5x）增强子元件序列，luciferase萤火虫荧光素酶报告基因，Amp抗性。

pNF-κB-Luc Enhancer element sequence

Enhancer element configuration :NF-κB (5×)

Enhancer element sequence: (TGGGGACTTTCCGC)5

The PathDetect in Vivo Signal Transduction Pathway cis-Reporting Systems are designed for simple, rapid, and convenient assessment of the invivo activation of signal transduction pathways. We have created a series ofinducible reporter plasmids that contain the luciferase reporter gene drivenby a basic promoter element (TATA box) plus a defined inducible cisenhancerelement (see Figure 2 in Appendix). These plasmids areparticularly suited for the in vivo readouts of signal transduction pathwayssince these enhancers are convergent points of many signal transductionpathways. These systems are useful in studying the in vivo effects of a newgene, growth factor, or drug candidate on the signaling pathway. Some ofthe reporters may also be used for cloning novel signal transduction genes,and may be useful for identifying drug candidates in a high-throughputformat.cis-Reporting SystemsThe PathDetect in vivo signal transduction pathway cis-reporting systemsinclude a reporter plasmid and a positive or negative control plasmid.Expression of the Photinus pyralis (firefly) luciferase gene in the reporterplasmid is controlled by a synthetic promoter that contains direct repeats ofthe transcription recognition sequences for the tumor suppressor proteinp53,1,2 the activator protein 1 (AP-1),1–3 the CCAAT-enhancer bindingprotein (C/EBP), the consensus sites for cyclic AMP response element(CRE),1–3 the retinoid X receptor response element (DR1/RXRE),4 thevitamin D receptor response element (DR3/VDR),4 the active thyroidhormone receptor response element (DR4/TR), the retinoic acid receptorresponse element (DR5/RARE),4,5 the early growth response factor 1(Egr-1),6 the interferon γ-activated sequence (GAS),1,2 the glucocorticoidresponse element (GRE),7 the interferon-stimulated response element(ISRE),1,2 the LPS IL-1β response element (LILRE),8,9 the nuclear factor ofactivated T cells (NFAT),1,2,10 the binding sites for nuclear factor κB(NF-κB),1–3 the serum response element (SRE),1–3 the serum response factor(SRF),1–3 or the TGF-β/Activin response element (TARE) (see Appendix:Plasmid Information).In addition to the luciferase reporter format, the CRE, AP-1, NF-kB andNFAT cis-reporting plasmids are available with an hrGFP reporter(humanized green fluorescent protein). With the hrGFP reporter, one candetect the signal without disrupting the cells, an advantage that isparticularly suitable for functional cloning and potentially for highthroughputscreening.When a reporter plasmid and an uncharacterized gene are cotransfected intomammalian cells, the activation of endogenous protein kinases initiated bythe cotransfected gene product will result in the activation of correspondingtrans-activators which in turn stimulate reporter expression (Figure 1).The effects of extracellular stimuli can also be studied with these systems.

Multiple Cloning Site Plasmid

Instead of repeats of a cis-acting DNA element, pLuc-MCS contains amultiple cloning site upstream from the luciferase gene and the TATA box(see Figure 3 in Appendix). Therefore, pLuc-MCS allows the insertion ofany cis-acting DNA element if a specific enhancer other than thosedescribed above is desired.Positive and Negative Control PlasmidsThe pCIS-CK negative control plasmid contains the luciferase reporter geneand does not contain any cis-acting DNA elements, thus serving as anegative control for all PathDetect cis-reporting plasmids. The negativecontrol is included with each of the following PathDetect cis-reportingsystems: C/EBP, DR1, DR3, DR4, DR5, Egr-1, GAS, GRE, ISRE, LILRE,NFAT, and TARE. For all other systems, the pCIS-CK negative controlplasmid is available separately.Each positive control plasmid expresses a kinase from the constitutiveCMV promoter that is ultimately responsible for activating transcription ofthe reporter from the enhancer element–TATA box region. A positivecontrol plasmid containing an appropriate kinase for a given pathway isprovided with each of the following cis-reporting systems: AP-1, CRE,NF-κB, p53, SRE, and SRF.

Map：

PathDetect in Vivo Signal Transduction Pathwaycis-Reporting Systems

MATERIALS PROVIDED

PathDetect AP-1 cis-Reporting System (Catalog #219073)Component Concentration QuantitypAP-1-Luc plasmid 1 μg/μl 50 μgpFC-MEKK positive control plasmid 25 ng/μl 5 μgPathDetect CRE cis-Reporting System (Catalog #219075)Component Concentration QuantitypCRE-Luc plasmid 1 μg/μl 50 μgpFC-PKA positive control plasmid 25 ng/μl 5 μgPathDetect NF-κB cis-Reporting System (Catalog #219077)Component Concentration QuantitypNF-κB-Luc plasmid 1 μg/μl 50 μgpFC-MEKK positive control plasmid 25 ng/μl 5 μgPathDetect SRE cis-Reporting System (Catalog #219079)Component Concentration QuantitypSRE-Luc plasmid 1 μg/μl 50 μgpFC-MEKK positive control plasmid 25 ng/μl 5 μgPathDetect SRF cis-Reporting System (Catalog #219081)Component Concentration QuantitypSRF-Luc plasmid 1 μg/μl 50 μgpFC-PKA positive control plasmid 25 ng/μl 5 μgPathDetect p53 cis-Reporting System (Catalog #219083)Component Concentration Quantityp53-Luc plasmid 1 μg/μl 50 μgpFC-p53 positive control plasmid 25 ng/μl 5 μgPathDetect ISRE cis-Reporting System (Catalog #219092)Component Concentration QuantitypISRE-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgPathDetect GAS cis-Reporting System (Catalog #219093)Component Concentration QuantitypGAS-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgRevision E.0 © Agilent Technologies, Inc. 2015. 2 PathDetect in Vivo Signal Transduction Pathway cis-Reporting SystemsPathDetect NFAT cis-Reporting System (Catalog #219094)Component Concentration QuantitypNFAT-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgPathDetect TARE cis-Reporting System (Catalog #219095)Component Concentration QuantitypTARE-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgPathDetect C/EBP cis-Reporting System (Catalog #240111)Component Concentration QuantitypC/EBP-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgPathDetect DR1 cis-Reporting System (Catalog #240113)Component Concentration QuantitypDR1-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgPathDetect DR3 cis-Reporting System (Catalog #240115)Component Concentration QuantitypDR3-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgPathDetect DR5 cis-Reporting System (Catalog #240119)Component Concentration QuantitypDR5-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgPathDetect Egr-1 cis-Reporting System (Catalog #240129)Component Concentration QuantitypEgr-1-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgPathDetect LILRE cis-Reporting System (Catalog #240131)Component Concentration QuantitypLILRE-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgPathDetect GRE cis-Reporting System (Catalog #240133)Component Concentration QuantitypGRE-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μg PathDetect in Vivo Signal Transduction Pathway cis-Reporting Systems 3PathDetect DR4 cis-Reporting System (Catalog #240135)Component Concentration QuantitypDR4-Luc plasmid 1 μg/μl 50 μgpCIS-CK negative control plasmid 1 μg/μl 50 μgPathDetect cis-Reporting PlasmidsComponent Concentration Quantity Catalog #pAP-1-Luc plasmid 1 μg/μl 50 μg 219074pCRE-Luc plasmid 1 μg/μl 50 μg 219076pNF-κB-Luc plasmid 1 μg/μl 50 μg 219078pSRE-Luc plasmid 1 μg/μl 50 μg 219080pSRF-Luc plasmid 1 μg/μl 50 μg 219082p53-Luc plasmid 1 μg/μl 50 μg 219085pISRE-Luc plasmid 1 μg/μl 50 μg 219089pGAS-Luc plasmid 1 μg/μl 50 μg 219091pNFAT-Luc plasmid 1 μg/μl 50 μg 219088pTARE-Luc plasmid 1 μg/μl 50 μg 240039pC/EBP-Luc plasmid 1 μg/μl 50 μg 240112pDR1-Luc plasmid 1 μg/μl 50 μg 240114pDR3-Luc plasmid 1 μg/μl 50 μg 240116pDR5-Luc plasmid 1 μg/μl 50 μg 240120pEgr-1-Luc plasmid 1 μg/μl 50 μg 240130pLILRE-Luc plasmid 1 μg/μl 50 μg 240132pGRE-Luc plasmid 1 μg/μl 50 μg 240134pDR4-Luc plasmid 1 μg/μl 50 μg 240136pAP-1-hrGFP plasmid 1 μg/μl 50 μg 240049pCRE-hrGFP plasmid 1 μg/μl 50 μg 240050pNF-κB-hrGFP plasmid 1 μg/μl 50 μg 240051pNFAT-hrGFP plasmid 1 μg/μl 50 μg 240053pLuc-MCS plasmid 1 μg/μl 50 μg 219087PathDetect cis-Reporting Control PlasmidsComponent Type of Control Concentration Quantity Catalog #pFC-p53 plasmid positive control plasmid recommended for use withp53-Luc plasmid 1 μg/μl 10 μg 219084pFC-MEKK plasmid positive control plasmid recommended for use withpNF-κB-Luc, pNF-κB-hrGFP, pAP-1-Luc,pAP-1-hrGFP, and pSRE-Luc plasmids 1 μg/μl 10 μg 219059pFC-PKA plasmid positive control plasmid recommended for use withpCRE-Luc, pCRE-hrGFP, and pSRF-Luc plasmids 1 μg/μl 10 μg 219071pCIS-CK plasmid negative control plasmid recommended for usewith all cis-reporting plasmids 1 μg/μl 50 μg 219090

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