ArticlePloS one2013
Construction of a stable replicating shuttle vector for Caldicellulosiruptor species: use for extending genetic methodologies to other members of this genus.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 50 papers.
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Who cites it
50 citing papers in PubMed, 76 citations in OpenAlex.
- A robust enzymatic reporter system for the extremely thermophilic anaerobic bacteriumFrontiers in microbiology · 2026Article
- An Initial Genome Editing Toolset for Caldimonas thermodepolymerans, the First Model of Thermophilic Polyhydroxyalkanoates Producer.Microbial biotechnology · 2025Article
- Metabolic engineering of Caldicellulosiruptor bescii for hydrogen production.Applied microbiology and biotechnology · 2024Review
- Sugar transport in thermophiles: Bridging lignocellulose deconstruction and bioconversion.Journal of industrial microbiology & biotechnology · 2024Review
- Hydrogen Production from Barley Straw and Miscanthus by the Hyperthermophilic Bacterium,Journal of microbiology and biotechnology · 2023Article
- Simultaneous expression of an endogenous spermidine synthase and a butanol dehydrogenase from Thermoanaerobacter pseudethanolicus in Clostridium thermocellum results in increased resistance to acetic acid and furans, increased ethanol production and an increase in thermotolerance.Biotechnology for biofuels and bioproducts · 2023Article
- A novel SfaNI-like restriction-modification system in Caldicellulosiruptor extents the genetic engineering toolbox for this genus.PloS one · 2022Article
- Genome-Scale Metabolic Model ofmSystems · 2021Article
- Approaches to genetic tool development for rapid domestication of non-model microorganisms.Biotechnology for biofuels · 2021Review
- Engineering the cellulolytic extreme thermophile Caldicellulosiruptor bescii to reduce carboxylic acids to alcohols using plant biomass as the energy source.Journal of industrial microbiology & biotechnology · 2020Article
- Insights into Thermophilic Plant Biomass Hydrolysis fromMicroorganisms · 2020Review
- The biology and biotechnology of the genus Caldicellulosiruptor: recent developments in 'Caldi World'.Extremophiles : life under extreme conditions · 2020Review
- Gene targets for engineering osmotolerance inBiotechnology for biofuels · 2020Article
- Genomic and physiological analyses reveal that extremely thermophilic Caldicellulosiruptor changbaiensis deploys uncommon cellulose attachment mechanisms.Journal of industrial microbiology & biotechnology · 2019Article
- An update of the suicide plasmid-mediated genome editing system in Corynebacterium glutamicum.Microbial biotechnology · 2019Article
- The thermophilic biomass-degrading bacteriumThe Journal of biological chemistry · 2019Article
- Expression of benzoyl-CoA metabolism genes in the lignocellulolytic host Caldicellulosiruptor bescii.AMB Express · 2019Article
- Heterologous co-expression of two β-glucanases and a cellobiose phosphorylase resulted in a significant increase in the cellulolytic activity of the Caldicellulosiruptor bescii exoproteome.Journal of industrial microbiology & biotechnology · 2019Article
- Rex in Caldicellulosiruptor bescii: Novel regulon members and its effect on the production of ethanol and overflow metabolites.MicrobiologyOpen · 2019Article
- Engineering redox-balanced ethanol production in the cellulolytic and extremely thermophilic bacterium,Metabolic engineering communications · 2018Article
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
The recalcitrance of plant biomass is the most important barrier to its economic conversion by microbes to products of interest. Thermophiles have special advantages for biomass conversion and members of the genus Caldicellulosiruptor are the most thermophilic cellulolytic microbes known. In this study, we report the construction of a replicating shuttle vector for Caldicellulosiruptor species based on pBAS2, the smaller of two native C. bescii plasmids. The entire plasmid was cloned into an E. coli cloning vector containing a pSC101 origin of replication and an apramycin resistance cassette for selection in E. coli. The wild-type C. bescii pyrF locus was cloned under the transcriptional control of the regulatory region of the ribosomal protein S30EA (Cbes2105), and the resulting vector was transformed into a new spontaneous deletion mutant in the pyrFA locus of C. bescii that allowed complementation with the pyrF gene alone. Plasmid DNA was methylated in vitro with a recently described cognate methyltransferase, M.CbeI, and transformants were selected for uracil prototrophy. The plasmid was stably maintained in low copy with selection but rapidly lost without selection. There was no evidence of DNA rearrangement during transformation and replication in C. bescii. A similar approach was used to screen for transformability of other members of this genus using M.CbeI to overcome restriction as a barrier and was successful for transformation of C. hydrothermalis, an attractive species for many applications. Plasmids containing a carbohydrate binding domain (CBM) and linker region from the C. bescii celA gene were maintained with selection and were structurally stable through transformation and replication in C. bescii and E. coli.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.