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Development of a synthetic oxytetracycline-inducible expression system for streptomycetes using de novo characterized genetic parts
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Wang, Weishan, Yang, Tongjian, Li, Yihong, Li, Shanshan, Yin, Shouliang, Styles, Kathryn, Corre, Christophe and Yang, Keqian (2016) Development of a synthetic oxytetracycline-inducible expression system for streptomycetes using de novo characterized genetic parts. ACS Synthetic Biology, 5 (7). pp. 765-773. doi:10.1021/acssynbio.6b00087
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WRAP_0380711-lf-160516-2016_wangyang_acssynbio.pdf - Accepted Version - Requires a PDF viewer. Download (2622Kb) | Preview |
Official URL: http://dx.doi.org/10.1021/acssynbio.6b00087
Abstract
Precise control of gene expression using exogenous factors is of great significance. To develop ideal inducible expression systems for streptomycetes, new genetic parts, oxytetracycline responsive repressor OtrR, operator otrO, and promoter otrBp from Streptomyces rimosus, were selected de novo and characterized in vivo and in vitro. OtrR showed strong affinity to otrO (KD = 1.7 × 10–10 M) and oxytetracycline induced dissociation of the OtrR/DNA complex in a concentration-dependent manner. On the basis of these genetic parts, a synthetic inducible expression system Potr* was optimized. Induction of Potr* with 0.01–4 μM of oxytetracycline triggered a wide-range expression level of gfp reporter gene in different Streptomyces species. Benchmarking Potr* against the widely used constitutive promoters ermE* and kasOp* revealed greatly enhanced levels of expression when Potr* was fully induced. Finally, Potr* was used as a tool to activate and optimize the expression of the silent jadomycin biosynthetic gene cluster in Streptomyces venezuelae. Altogether, the synthetic Potr* presents a new versatile tool for fine-tuning gene expression in streptomycetes.
| Item Type: | Journal Article | ||||||||
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| Subjects: | Q Science > QH Natural history > QH426 Genetics R Medicine > RM Therapeutics. Pharmacology |
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| Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) |
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| Library of Congress Subject Headings (LCSH): | Streptomycin, Gene expression, Synthetic biology, Antibiotics | ||||||||
| Journal or Publication Title: | ACS Synthetic Biology | ||||||||
| Publisher: | American Chemical Society | ||||||||
| ISSN: | 2161-5063 | ||||||||
| Official Date: | 15 July 2016 | ||||||||
| Dates: |
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| Volume: | 5 | ||||||||
| Number: | 7 | ||||||||
| Page Range: | pp. 765-773 | ||||||||
| DOI: | 10.1021/acssynbio.6b00087 | ||||||||
| Status: | Peer Reviewed | ||||||||
| Publication Status: | Published | ||||||||
| Access rights to Published version: | Restricted or Subscription Access | ||||||||
| Funder: | Guo jia zi ran ke xue ji jin wei yuan hui (China) [National Natural Science Foundation of China] (NSFC), China. Guo jia ke xue ji shu bu [Ministry of Science and Technology], Beijing Natural Science Foundation (BNSF), Zhongguo ke xue yuan [Chinese Academy of Sciences] (CAS) | ||||||||
| Grant number: | 31400034, 31570031 (NSFC), 2013CB734001, 5154032 (BNSF), 2016087 (CAS) |
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