A modular system for Clostridium shuttle plasmids
Abstract
Despite their medical and industrial importance, our basic understanding of the biology of the genus Clostridium is rudimentary in comparison to their aerobic counterparts in the genus Bacillus. A major contributing factor has been the comparative lack of sophistication in the gene tools available to the clostridial molecular biologist, which are immature, and in clear need of development. The transfer and maintenance of recombinant, replicative plasmids into various species of Clostridium has been reported, and several elements suitable as shuttle plasmid components are known. However, these components have to-date only been available in disparate plasmid contexts, and their use has not been broadly explored. Here we describe the specification, design and construction of a standardized modular system for Clostridium-Escherichia coli shuttle plasmids. Existing replicons and selectable markers were incorporated, along with a novel clostridial replicon. The properties of these components were compared, and the data allow researchers to identify combinations of components potentially suitable for particular hosts and applications. The system has been extensively tested in our laboratory, where it is utilized in all ongoing recombinant work. We propose that adoption of this modular system as a standard would be of substantial benefit to the Clostridium research community, whom we invite to use and contribute to the system.
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Modular Clostridium-E. coli shuttle plasmids developed by Heap et al. contain customizable components such as different Gram-positive and Gram-negative replicons and the traJ gene, and have been adopted for genetic manipulation work in several acetogens.
“The modular Clostridium-E. coli shuttle plasmids developed by Heap et al. (2009) contain customizable components such as different Gram-positive and Gram-negative replicons as well as the traJ gene (for conjugation), and it has been adopted for genetic manipulation work in several acetogens (Köpke and Liew, 2011; Leang et al., 2013; Ueki et al., 2014; Hoffmeister et al., 2016).”
From Gas Fermentation—A Flexible Platform for Commercial Scale Production of Low-Carbon-Fuels and Chemicals from Waste and Renewable Feedstocks (2016) · cites this paper as Heap et al., 2009
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Self-citation
- Authors
- Heap, J. T., Pennington, O. J., Cartman, S. T., & Minton, N. P.
- Affiliations
- University of Nottingham (GB)
- Corresponding author
- Minton, Nigel P.
- Venue
- Journal of Microbiological Methods
- Publisher
- Elsevier BV
- Published
- 2009-07
- Volume
- 78
- Issue
- 1
- Pages
- 79-85
- ISSN
- 0167-7012 (ISSN-L)
- OpenAlex
- W2047821263
- Cited by
- 489
- References
- 39
- Citations by year
- 2026: 27 · 2025: 26 · 2024: 35 · 2023: 38 · 2022: 38 · 2021: 42 · 2020: 29 · 2019: 33 · 2018: 35 · 2017: 20 · 2016: 35 · 2015: 28 · 2014: 28 · 2013: 31 · 2012: 18
- FWCI
- 8.58
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- Journal of Microbiological Methods (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Biotechnology and Biological Sciences Research Council (BB/D522289/1, BB/D522797/1, BB/F003390/1, BD/D522289/1)
- Staatssekretariat für Bildung, Forschung und Innovation
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Keywords
- Clostridium, shuttle plasmid, plasmid replicons, selectable marker, gene transfer, plasmid maintenance
- MeSH terms
- Clostridium, Conjugation, Genetic, Escherichia coli, Genetic Vectors, Plasmids, Gene Transfer Techniques
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