ArticleJournal of bacteriology2007
Disruption of a sugar transporter gene cluster in a hyperthermophilic archaeon using a host-marker system based on antibiotic resistance.
Article in Journal of bacteriology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 64 papers.
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Who cites it
64 citing papers in PubMed, 111 citations in OpenAlex.
- Removal and bypass of restriction modification systems increases transformation efficiency ofFrontiers in microbiology · 2026Article
- Pentose bisphosphate pathway can act in central metabolism for nucleoside-dependent growth ofApplied and environmental microbiology · 2025Article
- Distinct Patterns of Antibiotic Sensitivities in Ammonia-Oxidising Archaea.Environmental microbiology · 2025Article
- A self-transmissible plasmid from a hyperthermophile that facilitates genetic modification of diverse Archaea.Nature microbiology · 2023Article
- Biochemical and genetic examination of two aminotransferases from the hyperthermophilic archaeonFrontiers in microbiology · 2023Article
- A Novel NADP-Dependent Formate Dehydrogenase From the Hyperthermophilic ArchaeonFrontiers in microbiology · 2022Article
- Transformation Techniques for the Anaerobic Hyperthermophile Thermococcus kodakarensis.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Progress and Challenges in Archaeal Genetic Manipulation.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Structure and Function of Piezophilic HyperthermophilicInternational journal of molecular sciences · 2021Article
- High-efficiency transformation of archaea by direct PCR products with its application to directed evolution of a thermostable enzyme.Microbial biotechnology · 2021Article
- Thermococcus kodakarensis provides a versatile hyperthermophilic archaeal platform for protein expression.Methods in enzymology · 2021Article
- Thermophilic Chitinases: Structural, Functional and Engineering Attributes for Industrial Applications.Applied biochemistry and biotechnology · 2021Review
- A Structurally Novel Lipoyl Synthase in the Hyperthermophilic Archaeon Thermococcus kodakarensis.Applied and environmental microbiology · 2020Article
- Integration of large heterologous DNA fragments into the genome of Thermococcus kodakarensis.Extremophiles : life under extreme conditions · 2020Article
- An overview of 25 years of research on Thermococcus kodakarensis, a genetically versatile model organism for archaeal research.Folia microbiologica · 2020Review
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- A linear pathway for mevalonate production supports growth of Thermococcus kodakarensis.Extremophiles : life under extreme conditions · 2019Article
- Random mutagenesis of a hyperthermophilic archaeon identified tRNA modifications associated with cellular hyperthermotolerance.Nucleic acids research · 2019Article
- Induction of a Toxin-Antitoxin Gene Cassette under High Hydrostatic Pressure Enables Markerless Gene Disruption in the Hyperthermophilic ArchaeonApplied and environmental microbiology · 2019Article
4 more citing papers are in PubMed but not listed here.
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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
We have developed a gene disruption system in the hyperthermophilic archaeon Thermococcus kodakaraensis using the antibiotic simvastatin and a fusion gene designed to overexpress the 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase gene (hmg(Tk)) with the glutamate dehydrogenase promoter. With this system, we disrupted the T. kodakaraensis amylopullulanase gene (apu(Tk)) or a gene cluster which includes apu(Tk) and genes encoding components of a putative sugar transporter. Disruption plasmids were introduced into wild-type T. kodakaraensis KOD1 cells, and transformants exhibiting resistance to 4 microM simvastatin were isolated. The transformants exhibited growth in the presence of 20 microM simvastatin, and we observed a 30-fold increase in intracellular HMG-CoA reductase activity. The expected gene disruption via double-crossover recombination occurred at the target locus, but we also observed recombination events at the hmg(Tk) locus when the endogenous hmg(Tk) gene was used. This could be avoided by using the corresponding gene from Pyrococcus furiosus (hmg(Pf)) or by linearizing the plasmid prior to transformation. While both gene disruption strains displayed normal growth on amino acids or pyruvate, cells without the sugar transporter genes could not grow on maltooligosaccharides or polysaccharides, indicating that the gene cluster encodes the only sugar transporter involved in the uptake of these compounds. The Deltaapu(Tk) strain could not grow on pullulan and displayed only low levels of growth on amylose, suggesting that Apu(Tk) is a major polysaccharide-degrading enzyme in T. kodakaraensis.
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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.