ArticleExtremophiles : life under extreme conditions2015
Transposon mutagenesis of the extremely thermophilic bacterium Thermus thermophilus HB27.
Article in Extremophiles : life under extreme conditions, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 10 citations in OpenAlex.
- Construction of primary chassis cells with efficient protein expression in Thermus thermophilus.Microbial cell factories · 2025Article
- Genetic Manipulation of Neisseria gonorrhoeae and Commensal Neisseria Species.Current protocols · 2024Article
- Frequent Transposition of Multiple Insertion Sequences inFrontiers in microbiology · 2021Article
- Random mutagenesis of a hyperthermophilic archaeon identified tRNA modifications associated with cellular hyperthermotolerance.Nucleic acids research · 2019Article
- Development of the Multiple Gene Knockout System with One-Step PCR in Thermoacidophilic CrenarchaeonArchaea (Vancouver, B.C.) · 2017Article
- A reporter gene system for the precise measurement of promoter activity in Thermus thermophilus HB27.Extremophiles : life under extreme conditions · 2015Article
- Engineering the genome of Thermus thermophilus using a counterselectable marker.Journal of bacteriology · 2015Article
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
Thermus thermophilus is an extremely thermophilic bacterium that grows between 50 and 80 °C and is an excellent model organism not only for understanding life at high temperature but also for its biotechnological and industrial applications. Multiple molecular capabilities are available including targeted gene inactivation and the use of shuttle plasmids that replicate in T. thermophilus and Escherichia coli; however, the ability to disrupt gene function randomly by transposon insertion has not been developed. Here we report a detailed method of transposon mutagenesis of T. thermophilus HB27 based on the EZ-Tn5 system from Epicentre Biotechnologies. We were able to generate insertion mutations throughout the chromosome by in vitro transposition and transformation with mutagenized genomic DNA. We also report that an additional step, one that fills in single stranded gaps in donor DNA generated by the transposition reaction, was essential for successful mutagenesis. We anticipate that our method of transposon mutagenesis will enable further genetic development of T. thermophilus and may also be valuable for similar endeavors with other under-developed organisms.
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Registered trials
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