ArticleBMC biotechnology2012
Construction and transformation of a Thermotoga-E. coli shuttle vector.
Article in BMC biotechnology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Occurrence of Capnophilic Lactic Fermentation in the Hyperthermophilic Anaerobic BacteriumInternational journal of molecular sciences · 2022Article
- Adapted laboratory evolution of Thermotoga sp. strain RQ7 under carbon starvation.BMC research notes · 2022Article
- Improvement of COMicroorganisms · 2021Article
- Physiological, metabolic and biotechnological features of extremely thermophilic microorganisms.Wiley interdisciplinary reviews. Systems biology and medicine · 2017Review
- Development of a pyrE-based selective system for Thermotoga sp. strain RQ7.Extremophiles : life under extreme conditions · 2017Article
- Contribution of Pentose Catabolism to Molecular Hydrogen Formation by Targeted Disruption of Arabinose Isomerase (araA) in the Hyperthermophilic Bacterium Thermotoga maritima.Applied and environmental microbiology · 2017Article
- Complete genome sequence ofStandards in genomic sciences · 2017Article
- A Highly Thermostable Kanamycin Resistance Marker Expands the Tool Kit for Genetic Manipulation of Caldicellulosiruptor bescii.Applied and environmental microbiology · 2016Article
- Extremely thermophilic microorganisms as metabolic engineering platforms for production of fuels and industrial chemicals.Frontiers in microbiology · 2015Review
- Expression of Heterologous Cellulases in Thermotoga sp. Strain RQ2.BioMed research international · 2015Article
- A pipeline for completing bacterial genomes using in silico and wet lab approaches.BMC genomics · 2015Article
- Natural transformation of Thermotoga sp. strain RQ7.BMC biotechnology · 2014Article
- Thermophilic biohydrogen production: how far are we?Applied microbiology and biotechnology · 2013Review
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Authors and funding
3 authors.
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No grant is acknowledged in the PubMed record.
Abstract
backgroundThermotoga spp. are attractive candidates for producing biohydrogen, green chemicals, and thermostable enzymes. They may also serve as model systems for understanding life sustainability under hyperthermophilic conditions. A lack of genetic tools has hampered the investigation and application of these organisms. This study aims to develop a genetic transfer system for Thermotoga spp.
resultsMethods for preparing and handling Thermotoga solid cultures under aerobic conditions were optimized. A plating efficiency of ~50% was achieved when the bacterial cells were embedded in 0.3% Gelrite. A Thermotoga-E. coli shuttle vector pDH10 was constructed using pRQ7, a cryptic mini-plasmid found in T. sp. RQ7. Plasmid pDH10 was introduced to T. maritima and T. sp. RQ7 by electroporation and liposome-mediated transformation. Transformants were isolated, and the transformed kanamycin resistance gene (kan) was detected from the plasmid DNA extracts of the recombinant strains by PCR and was confirmed by restriction digestions. The transformed DNA was stably maintained in both Thermotoga and E. coli even without the selective pressure.
conclusionsThermotoga are transformable by multiple means. Recombinant Thermotoga strains have been isolated for the first time. A heterologous kan gene is functionally expressed and stably maintained in Thermotoga.
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