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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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Official URL: http://dx.doi.org/10.1021/acssynbio.6b00087

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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
Subjects: Q Science > QH Natural history > QH426 Genetics
R Medicine > RM Therapeutics. Pharmacology
Divisions: Faculty of Science > Chemistry
Faculty of Science > Life Sciences (2010- )
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:
DateEvent
15 July 2016Published
21 April 2016Available
21 April 2016Accepted
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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