Journal
PLANT SCIENCE
Volume 235, Issue -, Pages 14-24Publisher
ELSEVIER IRELAND LTD
DOI: 10.1016/j.plantsci.2015.02.014
Keywords
Potato (Solanum tuberosum L.); Promoter; Cold-inducible; C-repeat/dehydration-responsive element; Flanking sequence
Categories
Funding
- National Science Foundation of China [31171602]
- National High Technology Research and Development (863) Program of China [2007AA102188]
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Promoter activity is crucial for precise gene expression. Previously, a synthetic tuber-specific and cold-inducible promoter, pCL, containing a C-repeat/dehydration-responsive element (CRT/DRE) cassette and a tuber-specific fragment, was constructed in order to regulate cold-induced sweetening (CIS) in potatoes. However, the utility of pCL is limited due to its low activity. To improve its inducibility in response to low temperatures, we modified the CRT/DRE and flanking sequences. In particular, promoter activity was significantly improved by site-specific mutation of flanking sequences next to the core element (CCGAC) of CRT/DRE. We also inserted a modified CRT/DRE cassette into pCL; although this enhanced activity, it was not more effective than mutation of the flanking sequences. Indeed, up to 20-fold enhanced pCL activity could be achieved by replacing the CRT/DRE cassette in pCL with tandem repeats of two mutated CRT/DRE cassettes. This improvement was due to an enhanced affinity between the CRT/DRE cassette(s) and the StCBF1 transcription factor. Together, these data suggest that altering the structure of CRT/DRE can enhance CBF-related transcription complex formation and thus improve the activity of this cold-inducible promoter. (C) 2015 Elsevier Ireland Ltd. All rights reserved.
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