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Development of the Static and Dynamic Gene Expression Regulation Toolkit in Pseudomonas chlororaphisYue, Sheng-Jie; Zhou, Zheng; Huang, Peng; Wei, Yi-Chen; Zhan, Sheng-Xuan; Feng, Tong-Tong; Liu, Ji-Rui; Sun, Hao-Cheng; Han, Wei-Shang; Xue, Zhao-Long; Yan, Zi-Xin; Wang, Wei; Zhang, Xue-Hong; Hu, Hong-Bo
ACS synthetic biology, 03/2024, Letnik: 13, Številka: 3Journal Article
The advancement of metabolic engineering and synthetic biology has promoted in-depth research on the nonmodel microbial metabolism, and the potential of nonmodel organisms in industrial biotechnology is becoming increasingly evident. The nonmodel organism Pseudomonas chlororaphis is a safe plant growth promoting bacterium for the production of phenazine compounds; however, its application is seriously hindered due to the lack of an effective gene expression precise regulation toolkit. In this study, we constructed a library of 108 promoter-5′-UTR (PUTR) and characterized them through fluorescent protein detection. Then, 6 PUTRs with stable low, intermediate, and high intensities were further characterized by report genes lacZ encoding β-galactosidase from Escherichia coli K12 and phzO encoding PCA monooxygenase from P. chlororaphis GP72 and thus developed as a static gene expression regulation system. Furthermore, the stable and high-intensity expressed P MOK_RS0128085 UTR was fused with the LacO operator to construct an IPTG-induced plasmid, and a self-induced plasmid was constructed employing the high-intensity P MOK_RS0116635 UTR regulated by cell density, resulting in a dynamic gene expression regulation system. In summary, this study established two sets of static and dynamic regulatory systems for P. chlororaphis, providing an effective toolkit for fine-tuning gene expression and reprograming the metabolism flux.
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JCR | SNIP | JCR | SNIP | JCR | SNIP | JCR | SNIP |
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