Journal
PLANT AND CELL PHYSIOLOGY
Volume 54, Issue 10, Pages 1724-1735Publisher
OXFORD UNIV PRESS
DOI: 10.1093/pcp/pct115
Keywords
Circadian clock; Cyanobacteria; Riboswitch; Regulation of gene expression; Synechococcus elongatus PCC 7942; Theophylline
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Funding
- Ministry of Education, Culture, Sports, Science and Technology, Japan [24570054, 24370020, 24119510]
- Grants-in-Aid for Scientific Research [24570054, 24119510] Funding Source: KAKEN
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The cyanobacterium Synechococcus elongatus PCC 7942 is a major model species for studies of photosynthesis. It is are also a potential cell factory for the production of renewable biofuels and valuable chemicals. We employed engineered riboswitches to control translational initiation of target genes in this cyanobacterium. A firefly luciferase reporter assay revealed that three theophylline riboswitches performed as expected in the cyanobacterium. Riboswitch-E* exhibited very low leaky expression of luciferase and superior and dose-dependent on/off regulation of protein expression by theophylline. The maximum magnitude of the induction vs. basal level was similar to 190-fold. Furthermore, the induction level was responsive to a wide range of theophylline concentrations in the medium, from 0 to 2 mM, facilitating the fine-tuning of luciferase expression. We adapted this riboswitch to another gene regulation system, in which expression of the circadian clock kaiC gene product is controlled by the theophylline concentration in the culture medium. The results demonstrated that the adequately adjusted expression level of KaiC restored complete circadian rhythm in the kaiC-deficient arrhythmic mutant. This theophylline-dependent riboswitch system has potential for various applications as a useful genetic tool in cyanobacteria.
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