1. Minimizing endogenous cryptic plasmids to construct antibiotic-free expression systems for Escherichia coli Nissle 1917
- Author
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Siyan Zhou, Linlin Zhao, Wenjie Zuo, Yilin Zheng, Ping Zhang, Yanan Sun, Yang Wang, Guocheng Du, and Zhen Kang
- Subjects
Cryptic plasmids ,Antibiotic-free ,Gene expression ,Plasmid stability ,E. coli Nissle 1917 ,Biotechnology ,TP248.13-248.65 ,Biology (General) ,QH301-705.5 - Abstract
The probiotic bacterium Escherichia coli Nissle 1917 (EcN) holds significant promise for use in clinical and biological industries. However, the reliance on antibiotics to maintain plasmid-borne genes has overshadowed its benefits. In this study, we addressed this issue by engineering the endogenous cryptic plasmids pMUT1 and pMUT2. The non-essential elements were removed to create more stable derivatives pMUT1NR△ and pMUT2HBC△. Synthetic promoters by integrating binding motifs on sigma factors were further constructed and applied for expression of Bacteroides thetaiotaomicron heparinase III and the biosynthesis of ectoine. Compared to traditional antibiotic-dependent expression systems, our newly constructed antibiotic-free expression systems offer considerable advantages for clinical and synthetic biology applications.
- Published
- 2024
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