期刊
PLANT JOURNAL
卷 82, 期 4, 页码 707-716出版社
WILEY
DOI: 10.1111/tpj.12843
关键词
synthetic promoters; transcription activator-like effectors; orthogonality; metabolic engineering; transient assays; Nicotiana benthamiana; terpenoid; synthetic biology; plants; technical advance
资金
- DFG from the ERA-CAPS programme [TI 800/2-1]
- Leibniz-Institute of Plant Biochemistry, Halle, Germany
A library of synthetic promoters containing the binding site of a single designer transcription activator-like effector (dTALE) was constructed. The promoters contain a constant sequence, consisting of an 18-base long dTALE-binding site and a TATA box, flanked by degenerate sequences of 49 bases downstream and 19 bases upstream. Forty-three of these promoters were sequenced and tested in transient assays in Nicotiana benthamiana using a GUS reporter gene. The strength of expression of the promoters ranged from around 5% to almost 100% of the viral 35S promoter activity. We then demonstrated the utility of these promoters for metabolic engineering by transiently expressing three genes for the production of a plant diterpenoid in N.benthamiana. The simplicity of the promoter structure shows great promise for the development of genetic circuits, with wide potential applications in plant synthetic biology and metabolic engineering.
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