Cited by 92Bacterial Genetics and BiotechnologyCRISPR and Genetic EngineeringMicrobial Metabolic Engineering and Bioproduction

Engineering Clostridium strain to accept unmethylated DNA

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Dong, Hongjun, Zhang, Yanping, Dai, Zongjie, Li, Yin·2010·PLoS ONE

Abstract

It is difficult to genetically manipulate the medically and biotechnologically important genus Clostridium due to the existence of the restriction and modification (RM) systems. We identified and engineered the RM system of a model clostridial species, C. acetobutylicum, with the aim to allow the host to accept the unmethylated DNA efficiently. A gene CAC1502 putatively encoding the type II restriction endonuclease Cac824I was identified from the genome of C. acetobutylicum DSM1731, and disrupted using the ClosTron system based on group II intron insertion. The resulting strain SMB009 lost the type II restriction endonuclease activity, and can be transformed with unmethylated DNA as efficiently as with methylated DNA. The strategy reported here makes it easy to genetically modify the clostridial species using unmethylated DNA, which will help to advance the understanding of the clostridial physiology from the molecular level.

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  1. [30]Heap J Pennington O Cartman S Carter G Minton N 2007 The ClosTron: a universal gene knock-out system for the genus Clostridium. J Microbiol Meth 70 452 464 17658189

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Authors
Dong, H., Zhang, Y., Dai, Z., & Li, Y.
Affiliations
  • Chinese Academy of Sciences (CN)
  • University of Science and Technology of China (CN)
Corresponding author
Li, Yin
Venue
PLoS ONE
Published
2010-02-09
Volume
5
Issue
2
Pages
e9038
ISSN
1932-6203 (ISSN-L)
OpenAlex
W2031351113
Cited by
91
References
51
Citations by year
2026: 1 · 2025: 4 · 2024: 3 · 2023: 1 · 2022: 3 · 2021: 6 · 2020: 1 · 2019: 6 · 2018: 4 · 2017: 4 · 2016: 6 · 2015: 4 · 2014: 13 · 2013: 9 · 2012: 10
FWCI
6.29
Type
journal article
Language
en
Open access
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Chinese Academy of Sciences
  • National Key Research and Development Program of China (2007CB707803, 973 Project)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
Keywords
Clostridium acetobutylicum, restriction-modification systems, type II restriction endonuclease, DNA methylation, genome editing
MeSH terms
Bacterial Proteins, Clostridium, DNA, Genetic Engineering, Mutation, Plasmids, Transformation, Genetic, Deoxyribonucleases, Type II Site-Specific, DNA Restriction-Modification Enzymes, DNA Methylation, Clostridium acetobutylicum

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