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KMID : 0364219960390040378
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Korean Journal of Zoology
1996 Volume.39 No. 4 p.378 ~ p.386
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Versatile Luciferase Reporter Plasmids for Transcription Studies in Diverse Eukaryotic Cells
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Cho Yong-Seok
Han Dong-Uck
Baek Gum-Hee
Park Seung-Pil
Yoon Sang-Soon
Lim Woon-Ki
Kim Chong-Rak
Kim Han-Do
Kang Ho-Sung
Han Kyu-Hyung
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Abstract
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We have developed a couple of new luciferase reporter plasmida with very low background reporter actlvltlea. One can be used to measure the promoter strength, after insertion of some promoter fragment into the reporter plasmid, and the other, with very low basal promoter actlvltlea, allis In studying eukaryotic transcriptional regulators. The latter reporter plasmid contains such cli elements as a 17 nucleotide long inftlator, Spl.blndIng sftes, GAL4 binding sltea, and bInding sitea for a certain Drosophila homeodomain proteins. In an attempt to construct an improved reporter plaimid by fadlltating transcriptional termination and minimizing any interference by cryptic promoters which may be preaent in the reporter pleamld DNA, we have inserted transsrlptional termination-related signals, a three tandem repeat of SV4O polyadenylatlon signal (AAA) and the putative transcrtptional termination signal (UMS) of the mouse c-mos gene, Into just upstream of the initIator, and the promoter actlvitiea were measured by a transIent expression assay employing the Drosophila Schneider line 2 cells. As expected, the basal promoter activitIes decreased maximally when both transcription termination related elements were inserted. Moreover, the reporter plasmld with the two elements allowed more sensitive measurement of transcriptional activation than the reporter piasmid without them. Theae reporter plasmids can be used for studying transcriptional regulators of higher organisms Including mammals as well as Droiophlla melanogaiter.
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KEYWORD
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Reporter, Luciferase, Transcription, Initator, Drosophila
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