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Abstract
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A novel circadian phenotype based on firefly luciferase expression in transgenic plants.

A J Millar, S R Short, N H Chua, S A Kay
A J Millar
Laboratory of Plant Molecular Biology, Rockefeller University, New York, New York 10021-6399.
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S R Short
Laboratory of Plant Molecular Biology, Rockefeller University, New York, New York 10021-6399.
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N H Chua
Laboratory of Plant Molecular Biology, Rockefeller University, New York, New York 10021-6399.
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S A Kay
Laboratory of Plant Molecular Biology, Rockefeller University, New York, New York 10021-6399.
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Published September 1992. DOI: https://doi.org/10.1105/tpc.4.9.1075

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Abstract

A 320-bp fragment of the Arabidopsis cab2 promoter is sufficient to mediate transcriptional regulation by both phytochrome and the circadian clock. We fused this promoter fragment to the firefly luciferase (Luc) gene to create a real-time reporter for regulated gene expression in intact plants. Cab2::Luc transcript accumulated in the expected patterns and luciferase activity was closely correlated to cab2::Luc mRNA abundance in both etiolated and green seedlings. The concentration of the bulk of luciferase protein did not reflect these patterns but maintained a relatively constant level, implying that a post-translational mechanism(s) leads to the high-amplitude regulation of luciferase activity. We used a low-light video imaging system to establish that luciferase bioluminescence in vivo accurately reports the temporal and spatial regulation of cab2 transcription in single seedlings. The unique qualities of the firefly luciferase system allowed us to monitor regulated gene expression in real time in individual multicellular organisms. This noninvasive marker for temporal regulation at the molecular level constitutes a circadian phenotype, which may be used to isolate mutants in the circadian clock.

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A novel circadian phenotype based on firefly luciferase expression in transgenic plants.
A J Millar, S R Short, N H Chua, S A Kay
The Plant Cell Sep 1992, 4 (9) 1075-1087; DOI: 10.1105/tpc.4.9.1075

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A novel circadian phenotype based on firefly luciferase expression in transgenic plants.
A J Millar, S R Short, N H Chua, S A Kay
The Plant Cell Sep 1992, 4 (9) 1075-1087; DOI: 10.1105/tpc.4.9.1075
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Vol. 4, Issue 9
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