Abstract
We show that firefly luciferase is a stable protein when expressed at 25 °C in Neurospora, which limits its use as transcription reporter. We created a short-lived luciferase by fusing a PEST signal to its C-terminus (LUC-PEST) and applied the LUC-PEST reporter system to record in vivo transcription dynamics associated with the Neurospora circadian clock and its blue-light photosensory system over the course of several days. We show that the tool is suitable to faithfully monitor rapid, but also subtle changes in transcription in a medium to high throughput format.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Circadian Rhythm / genetics*
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Fungal Proteins / genetics*
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Light*
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Luciferases / genetics
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Luciferases / metabolism*
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Luminescent Measurements
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Molecular Sequence Data
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Neurospora / genetics*
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Neurospora / metabolism
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Polymerase Chain Reaction
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Transcription, Genetic*
Substances
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FRQ protein, Neurospora crassa
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Fungal Proteins
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Luciferases
Grants and funding
This work was supported by grants from the Deutsche Forschungsgemeinschaft: DFG DI 1874/1-1 to AD (
http://www.dfg.de). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.