Abstract
Development of strong promoters is of growing interest in the field of biotechnology and synthetic biology. Here we present a protocol for the construction of strong prokaryotic promoters that can be recognized by designated multiple sigma factors by interlocking their cognate binding motifs on DNA strands. Strong and stress responsive promoters for Escherichia coli and Bacillus subtilis have been created following the presented protocol. Customized promoters could be easily developed for fine-tuning gene expression or overproducing enzymes with prokaryotic cell factories.
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Acknowledgments
This work was financially supported by the National Natural Science Foundation of China (31970085, 32000058), the Jiangsu Province Natural Science Fund for Distinguished Young Scholars (BK20200025), the National Key R&D program of China (2018YFA0901400), a grant from the Key Technologies R&D Program of Jiangsu Province (BE2019630), and the National First-class Discipline Program of Light Industry Technology and Engineering (LITE2018-16).
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Zhang, Y., Wang, Y., Li, J., Wang, C., Du, G., Kang, Z. (2022). Construction of Strong Promoters by Assembling Sigma Factor Binding Motifs. In: Currin, A., Swainston, N. (eds) Directed Evolution. Methods in Molecular Biology, vol 2461. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2152-3_9
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DOI: https://doi.org/10.1007/978-1-0716-2152-3_9
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