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An operator-based expression toolkit for Bacillus subtilis enables fine-tuning of gene expression and biosynthetic pathway regulation.

ABSTRACT: SignificanceA gene regulatory system is an important tool for the engineering of biosynthetic pathways of organisms. Here, we report the development of an inducible-ON/OFF regulatory system using a malO operator as a key element. We identified and modulated sequence, position, numbers, and spacing distance of malO operators, generating a series of activating or repressive promoters with tunable strength. The stringency and robustness are both guaranteed in this system, a maximal induction factor of 790-fold was achieved, and nine proteins from different organisms were expressed with high yields. This system can be utilized as a gene switch, promoter enhancer, or metabolic valve in synthetic biology applications. This operator-based engineering strategy can be employed for developing similar regulatory systems in different microorganisms.

SUBMITTER: Fu G 

PROVIDER: S-EPMC8931375 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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An operator-based expression toolkit for <i>Bacillus subtilis</i> enables fine-tuning of gene expression and biosynthetic pathway regulation.

Fu Gang G   Yue Jie J   Li Dandan D   Li Yixin Y   Lee Sang Yup SY   Zhang Dawei D  

Proceedings of the National Academy of Sciences of the United States of America 20220309 11

SignificanceA gene regulatory system is an important tool for the engineering of biosynthetic pathways of organisms. Here, we report the development of an inducible-ON/OFF regulatory system using a <i>malO</i> operator as a key element. We identified and modulated sequence, position, numbers, and spacing distance of <i>malO</i> operators, generating a series of activating or repressive promoters with tunable strength. The stringency and robustness are both guaranteed in this system, a maximal in  ...[more]