期刊
METABOLIC ENGINEERING
卷 19, 期 -, 页码 98-106出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2013.07.006
关键词
Synthetic promoter; Actinomycetes; GusA; RppA; Transcriptomic analysis; RNA-Seq; Erme promoter
资金
- ERC [281623]
- European Research Council (ERC) [281623] Funding Source: European Research Council (ERC)
We developed a synthetic promoter library for actinomycetes based on the 10 and 35 consensus sequences of the constitutive and widely used ermEp1 promoter. The sequences located upstream, in between and downstream of these consensus sequences were randomised using degenerate primers and cloned into an integrative plasmid upstream of the gusA reporter gene. Using this system, we created promoters with strengths ranging from 2% to 319% compared with ermEp1. The strongest synthetic promoter was used in a proof-of-principle approach to achieve the overexpression of a natural type III polyketide synthase. We observed high correlation between the number of gusA reporter gene RNA-Seq reads and the GusA reporter protein activity, indicating that GusA is indeed a transcription-level reporter system. (C) 2013 Elsevier Inc. All rights reserved.
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