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dc.contributor.authorGhazawi, I.en_US
dc.contributor.authorCutler, S.en_US
dc.contributor.authorKelly, Paulineen_US
dc.contributor.authorMellick, Alberten_US
dc.contributor.authorRalph, Stephenen_US
dc.contributor.editorGanes C. Sen and Thomas A. Hamiltonen_US
dc.date.accessioned2017-05-03T14:14:56Z
dc.date.available2017-05-03T14:14:56Z
dc.date.issued2005en_US
dc.identifier.issn10799907en_US
dc.identifier.doi10.1089/jir.2005.25.92en_US
dc.identifier.urihttp://hdl.handle.net/10072/4725
dc.description.abstractLuciferase reporter constructs are widely used for analysis of gene regulation when characterizing promoter and enhancer elements. We report that the recently developed codon-modified Renilla luciferase construct included as an internal standard for cotransfection must be used with great caution with respect to the amount of DNA transfected. Also, the dual-luciferase reporter vectors encoding Photinus pyralis firefly or Renilla reniformis luciferase showed a linear increase in dose-response with increasing amounts of transfected DNA, but at higher levels of transfected DNA, a reduction in expressed levels of luciferase activity resulted. In addition, treatment with type I interferon (IFN) was found to significantly reduce levels of P. pyralis firefly and Renilla luciferase activity. In contrast, cells transfected with a green fluorescent protein (GFP) reporter construct showed no significant IFN-associated change. The reduction in luciferase activity resulting from IFN treatment was not due to IFN-mediated cytotoxicity, as no change in cellular propidium iodide (PI) staining was observed by flow cytometry. IFN treatment did not alter the levels of firefly luciferase activity in cell culture supernatants or the luciferase mRNA levels determined by quantitative real-time RT-PCR analysis. Based on these results, it is probable that the IFN-induced reduction in levels of luciferase activity detected in reporter assays occurs via a posttranscriptional mechanism. Thus, it is important to be aware of these complications when using luciferase reporter systems in general or for analyzing cytokine-mediated responsive regulation of target genes, particularly by the type I IFNs.en_US
dc.description.peerreviewedYesen_US
dc.description.publicationstatusYesen_US
dc.languageEnglishen_US
dc.language.isoen_US
dc.publisherMary Ann Liebert, Incen_US
dc.publisher.placeUSAen_US
dc.relation.ispartofstudentpublicationNen_US
dc.relation.ispartofpagefrom92en_US
dc.relation.ispartofpageto102en_US
dc.relation.ispartofissue2en_US
dc.relation.ispartofjournalJournal of Interferon and Cytokine Researchen_US
dc.relation.ispartofvolume25en_US
dc.rights.retentionYen_US
dc.subject.fieldofresearchcode270199en_US
dc.titleInhibitory Effects Associated with Use of Modified Photinus pyralis and Renilla reniformis Luciferase Vectors in Dual Reporter Assays and Implications for Analysis of ISGs.en_US
dc.typeJournal articleen_US
dc.type.descriptionC1 - Peer Reviewed (HERDC)en_US
dc.type.codeC - Journal Articlesen_US
gro.date.issued2015-05-12T05:11:12Z
gro.hasfulltextNo Full Text


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