Open AccessLight-Dependent and Circadian Transcription Dynamics In Vivo Recorded with a Destabilized Luciferase Reporter in Neurospora
Author(s) -
François Cesbron,
Michael Brunner,
Axel Diernfellner
Publication year - 2013
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0083660
Subject(s) - luciferase , neurospora , circadian clock , transcription factor , transcription (linguistics) , biology , in vivo , circadian rhythm , firefly protocol , bioluminescence , microbiology and biotechnology , genetics , biochemistry , gene , evolutionary biology , neuroscience , transfection , linguistics , philosophy , mutant , neurospora crassa
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.