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Characterization and implications of prokaryotic ribosome-binding sites across species

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Abstract

The ribosome-binding site (RBS) in the 5′ untranslated region is recognized by 16S rRNA to start translation and is an essential element of the gene expression system. RBSs have been widely applied in regulating gene expression in various scenarios, including Gram-negative or Gram-positive bacteria. Here, we first rationally designed and constructed an RBS mutant library containing 66 RBSs. The strength of these RBSs in E. coli and C. glutamicum was characterized individually. The RBS strength spanned about 200 and 15 times in the two species, respectively. The strength of RBSs in C. glutamicum was generally lower than that of in E. coli. A total of 18 RBSs showed similar strength (within twofold differences) between the species in our study, and the correlation analysis of the strength of RBSs between E. coli and C. glutamicum (R2 = 0.7483) revealed that these RBSs can be used across species. The sequence analysis revealed that the RBS region with two Ts stated was beneficial for RBS to function cross-species. The RBS characterized here can be used to precisely regulate gene expression in both hosts, and the characteristics of cross-species RBSs provide basic information for RBS rational design.

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Acknowledgements

This work was financially supported by the National Key Research and Development Program of China (2018YFA0900300) and the National Natural science foundation of China (32171421).

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Authors and Affiliations

  1. Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China

    Yanting Duan, Xiaojuan Zhang, Weiji Zhai, Jinpeng Zhang, Jiawei Ren, Guoqiang Xu & Zhenghong Xu

  2. National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi, China

    Yanting Duan, Xiaojuan Zhang, Weiji Zhai, Jinpeng Zhang, Jiawei Ren, Guoqiang Xu & Zhenghong Xu

  3. School of Life Science and Health Engineering, Jiangnan University, Wuxi, 214122, China

    Xiaomei Zhang & Jinsong Shi

  4. Jiangsu Engineering Research Center for Bioactive Products Processing Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China

    Xiaomei Zhang & Jinsong Shi

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  1. Yanting Duan
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  2. Xiaojuan Zhang
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  3. Weiji Zhai
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  4. Jinpeng Zhang
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  8. Jinsong Shi
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Correspondence to Xiaojuan Zhang or Zhenghong Xu.

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Duan, Y., Zhang, X., Zhai, W. et al. Characterization and implications of prokaryotic ribosome-binding sites across species. Syst Microbiol and Biomanuf 2, 676–684 (2022). https://doi.org/10.1007/s43393-022-00094-8

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  • DOI: https://doi.org/10.1007/s43393-022-00094-8

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