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OtherMOLECULAR BIOLOGY AND GENE REGULATION
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In Vitro Analysis of Light-Induced Transcription in the Wheat psbD/C Gene Cluster Using Plastid Extracts from Dark-Grown and Short-Term-Illuminated Seedlings

T. Wada, Y. Tunoyama, T. Shiina, Y. Toyoshima
T. Wada
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Y. Tunoyama
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T. Shiina
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Y. Toyoshima
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Published April 1994. DOI: https://doi.org/10.1104/pp.104.4.1259

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Abstract

We describe a plastid in vitro transcription system that reflects characteristic features of the light-regulated transcription observed in vivo. Multiple transcripts of the wheat (Triticum aestivum) psbD/C gene cluster comprise six distinct 5[prime] ends including four transcription initiation sites designated as D/C-1 through D/C-4. Transcripts from one particular site, D/C-3, were found to be conspicuously enhanced in abundance after 4 h of illumination in vivo. The plastid extract prepared from 5-d-old dark-grown wheat seedlings was capable of transcribing from the D/C-2 and D/C-4 sites in vitro but had almost no transcription activity from the light-responsive D/C-3 site (the D/C-1 site was not examined). The plastid extract from 4-h-illuminated seedlings initiated transcription from the light-responsive site (D/C-3). Transcription from the D/C-2 and D/C-4 sites was not enhanced by using the extract from 4-h-illuminated seedlings, indicative of specific activation of the light-responsive promoter on the D/C-3 site by the extract from 4-h-illuminated seedlings. The plastid extract from 4-h-illuminated seedlings was divided into two fractions on a heparin-Sepharose column, into which the light-induced component(s) responsible for activation of the D/C-3 promoter and RNA polymerase were separated. The fraction containing the component(s) activating the D/C-3 promoter induced the transcription activity from the D/C-3 site in the plastid extract from dark-grown seedlings. It is concluded that the plastid extract from 4-h-illuminated seedlings contains some light-regulatory component(s) that activate specifically the light-responsive promoter.

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In Vitro Analysis of Light-Induced Transcription in the Wheat psbD/C Gene Cluster Using Plastid Extracts from Dark-Grown and Short-Term-Illuminated Seedlings
T. Wada, Y. Tunoyama, T. Shiina, Y. Toyoshima
Plant Physiology Apr 1994, 104 (4) 1259-1267; DOI: 10.1104/pp.104.4.1259

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In Vitro Analysis of Light-Induced Transcription in the Wheat psbD/C Gene Cluster Using Plastid Extracts from Dark-Grown and Short-Term-Illuminated Seedlings
T. Wada, Y. Tunoyama, T. Shiina, Y. Toyoshima
Plant Physiology Apr 1994, 104 (4) 1259-1267; DOI: 10.1104/pp.104.4.1259
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Plant Physiology
Vol. 104, Issue 4
Apr 1994
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  • Genes Galore: A Summary of Methods for Accessing Results from Large-Scale Partial Sequencing of Anonymous Arabidopsis cDNA Clones
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  • Molecular Genetics of the Maize (Zea mays L.) Aspartate Kinase-Homoserine Dehydrogenase Gene Family
Show more Molecular Biology and Gene Regulation

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