ArticleBiotechnology for biofuels2012
Dcm methylation is detrimental to plasmid transformation in Clostridium thermocellum.
Article in Biotechnology for biofuels, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed.
- Effects of heterologous expression of glycolysis enzymes on product formation inMetabolic engineering communications · 2026Article
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- Epigenetic regulation in prokaryotes: transcriptional and phenotypic outcomes of DNA methyltransferase activity.FEMS microbiology reviews · 2026Review
- EngineeringbioRxiv : the preprint server for biology · 2025Article
- Thermophilic site-specific recombination system for rapid insertion of heterologous DNA into the Clostridium thermocellum chromosome.Journal of industrial microbiology & biotechnology · 2024Article
- Pyrophosphate-free glycolysis in Clostridium thermocellum increases both thermodynamic driving force and ethanol titers.Biotechnology for biofuels and bioproducts · 2024Article
- Expression and characterization of monofunctional alcohol dehydrogenase enzymes inMetabolic engineering communications · 2024Article
- Generation of tools for expression and purification of the phage-encoded Type I restriction enzyme inhibitor, Ocr.Microbiology (Reading, England) · 2024Article
- Extremophiles in a changing world.Extremophiles : life under extreme conditions · 2024Review
- Methylome-dependent transformation ofmBio · 2023Article
- The Roles of Nicotinamide Adenine Dinucleotide Phosphate Reoxidation and Ammonium Assimilation in the Secretion of Amino Acids as Byproducts of Clostridium thermocellum.Applied and environmental microbiology · 2023Article
- Increasing the Thermodynamic Driving Force of the Phosphofructokinase Reaction inApplied and environmental microbiology · 2022Article
- Breaking the Restriction Barriers and Applying CRISPRi as a Gene Silencing Tool inMicroorganisms · 2022Article
- Thermophilic whole-cell degradation of polyethylene terephthalate using engineered Clostridium thermocellum.Microbial biotechnology · 2021Article
- Approaches to genetic tool development for rapid domestication of non-model microorganisms.Biotechnology for biofuels · 2021Review
- Rational development of transformation in Clostridium thermocellum ATCC 27405 via complete methylome analysis and evasion of native restriction-modification systems.Journal of industrial microbiology & biotechnology · 2019Article
- Article
- Construction of consolidated bio-saccharification biocatalyst and process optimization for highly efficient lignocellulose solubilization.Biotechnology for biofuels · 2019Article
- Single mutation at a highly conserved region of chloramphenicol acetyltransferase enables isobutyl acetate production directly from cellulose byBiotechnology for biofuels · 2019Article
- Development of a metabolic pathway transfer and genomic integration system for the syngas-fermenting bacteriumBiotechnology for biofuels · 2019Article
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4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
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backgroundIndustrial production of biofuels and other products by cellulolytic microorganisms is of interest but hindered by the nascent state of genetic tools. Although a genetic system for Clostridium thermocellum DSM1313 has recently been developed, available methods achieve relatively low efficiency and similar plasmids can transform C. thermocellum at dramatically different efficiencies.
resultsWe report an increase in transformation efficiency of C. thermocellum for a variety of plasmids by using DNA that has been methylated by Escherichia coli Dam but not Dcm methylases. When isolated from a dam+dcm+E. coli strain, pAMG206 transforms C. thermocellum 100-fold better than the similar plasmid pAMG205, which contains an additional Dcm methylation site in the pyrF gene. Upon removal of Dcm methylation, transformation with pAMG206 showed a four- to seven-fold increase in efficiency; however, transformation efficiency of pAMG205 increased 500-fold. Removal of the Dcm methylation site from the pAMG205 pyrF gene via silent mutation resulted in increased transformation efficiencies equivalent to that of pAMG206. Upon proper methylation, transformation efficiency of plasmids bearing the pMK3 and pB6A origins of replication increased ca. three orders of magnitude.
conclusionsE. coli Dcm methylation decreases transformation efficiency in C. thermocellum DSM1313. The use of properly methylated plasmid DNA should facilitate genetic manipulation of this industrially relevant bacterium.
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