ArticleApplied and environmental microbiology2010
Shuttle vector-based transformation system for Pyrococcus furiosus.
Article in Applied and environmental microbiology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 48 papers.
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Who cites it
48 citing papers in PubMed, 81 citations in OpenAlex.
- Promoter contacts by TFB and TFEα during archaeal transcription initiation.Transcription · 2026Article
- Removal and bypass of restriction modification systems increases transformation efficiency ofFrontiers in microbiology · 2026Article
- The Thermococcales as a model system: historical perspectives and emerging tools.Journal of bacteriology · 2025Review
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- Structural basis of archaeal RNA polymerase transcription elongation and Spt4/5 recruitment.Nucleic acids research · 2024Article
- Archaea as a Model System for Molecular Biology and Biotechnology.Biomolecules · 2023Review
- The transcriptional regulator EarA and intergenic terminator sequences modulate archaellation inFrontiers in microbiology · 2023Article
- Antibacterial activities of coumarin-3-carboxylic acid againstFrontiers in microbiology · 2023Article
- Hyper-stimulation of Pyrococcus furiosus CRISPR DNA uptake by a self-transmissible plasmid.Extremophiles : life under extreme conditions · 2022Article
- Thermophilic Chitinases: Structural, Functional and Engineering Attributes for Industrial Applications.Applied biochemistry and biotechnology · 2021Review
- Insights into gene expression changes under conditions that facilitate horizontal gene transfer (mating) of a model archaeon.Scientific reports · 2020Article
- Direct binding of TFEα opens DNA binding cleft of RNA polymerase.Nature communications · 2020Article
- CopR, a Global Regulator of Transcription to Maintain Copper Homeostasis inFrontiers in microbiology · 2020Article
- Inward-facing conformation of a multidrug resistance MATE family transporter.Proceedings of the National Academy of Sciences of the United States of America · 2019Article
- A Genetic System forFrontiers in microbiology · 2019Article
- Displacement of the transcription factor B reader domain during transcription initiation.Nucleic acids research · 2018Article
- Biotechnology of extremely thermophilic archaea.FEMS microbiology reviews · 2018Review
- Article
- The Transcriptional Regulator TFB-RF1 Activates Transcription of a Putative ABC Transporter inFrontiers in microbiology · 2018Article
- Role of free DNA ends and protospacer adjacent motifs for CRISPR DNA uptake in Pyrococcus furiosus.Nucleic acids research · 2017Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Pyrococcus furiosus is a model organism for analyses of molecular biology and biochemistry of archaea, but so far no useful genetic tools for this species have been described. We report here a genetic transformation system for P. furiosus based on the shuttle vector system pYS2 from Pyrococcus abyssi. In the redesigned vector, the pyrE gene from Sulfolobus was replaced as a selectable marker by the 3-hydroxy-3-methylglutaryl coenzyme A reductase gene (HMG-CoA) conferring resistance of transformants to the antibiotic simvastatin. Use of this modified plasmid resulted in the overexpression of the HMG-CoA reductase in P. furiosus, allowing the selection of strains by growth in the presence of simvastatin. The modified shuttle vector replicated in P. furiosus, but the copy number was only one to two per chromosome. This system was used for overexpression of His(6)-tagged subunit D of the RNA polymerase (RNAP) in Pyrococcus cells. Functional RNAP was purified from transformed cells in two steps by Ni-NTA and gel filtration chromatography. Our data provide evidence that expression of transformed genes can be controlled from a regulated gluconeogenetic promoter.
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