Genes and Genetic Manipulation in Thermus thermophilus

Oshima, Tairo

doi:10.1007/978-1-4615-1831-0_7

Cited by 3 publications

(2 citation statements)

References 73 publications

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“…Some of the current gaps in the technology are obvious: ‘plug-and-play’ laboratory expression hosts for the major groups of extremophiles (e.g., thermophilic bacteria and archaea, psychrophiles, acidophiles, halophiles), where intracellular expression is highly impacted by the extracellular environment (temperature, pH, salt), are only available for a limited range of organisms. Probably the first thermophilic organism to be genetically manipulated was Thermus thermophiles (Oshima 1995 ), and this organism has now been engineered as a thermophilic host for the over-expression of proteins that cannot be expressed in mesophilic systems (De Rose et al 2021a ). However, the very high G + C content of the DNA of this organism generally means that genes to be expressed in this host system have to be synthesized.…”

Section: Gene Discoverymentioning

confidence: 99%

Extremophiles in a changing world

Cowan,

Albers,

Antranikian

et al. 2024

Extremophiles

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Extremophiles and their products have been a major focus of research interest for over 40 years. Through this period, studies of these organisms have contributed hugely to many aspects of the fundamental and applied sciences, and to wider and more philosophical issues such as the origins of life and astrobiology. Our understanding of the cellular adaptations to extreme conditions (such as acid, temperature, pressure and more), of the mechanisms underpinning the stability of macromolecules, and of the subtleties, complexities and limits of fundamental biochemical processes has been informed by research on extremophiles. Extremophiles have also contributed numerous products and processes to the many fields of biotechnology, from diagnostics to bioremediation. Yet, after 40 years of dedicated research, there remains much to be discovered in this field. Fortunately, extremophiles remain an active and vibrant area of research. In the third decade of the twenty-first century, with decreasing global resources and a steadily increasing human population, the world’s attention has turned with increasing urgency to issues of sustainability. These global concerns were encapsulated and formalized by the United Nations with the adoption of the 2030 Agenda for Sustainable Development and the presentation of the seventeen Sustainable Development Goals (SDGs) in 2015. In the run-up to 2030, we consider the contributions that extremophiles have made, and will in the future make, to the SDGs.

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Section: Gene Discoverymentioning

confidence: 99%

Extremophiles in a changing world

Cowan,

Albers,

Antranikian

et al. 2024

Extremophiles

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“…We have, therefore, initiated a genetic dissection of ribosomes of the extreme thermophile Thermus thermophilus. Despite the promise of Thermus as a genetic system (16,21), the lack of selectable genetic markers such as transposons or antibiotic resistance mutations makes the mapping of mutations to ribosomal protein and RNA genes problematic. Here we report the isolation of a 23S rRNA mutation conferring resistance to the antibiotics erythromycin and lincomycin.…”

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confidence: 99%

Spontaneous Erythromycin Resistance Mutation in a 23S rRNA Gene, rrlA , of the Extreme Thermophile Thermus thermophilus IB-21

2001

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Spontaneous, erythromycin-resistant mutants of Thermus thermophilus IB-21 were isolated and found to carry the mutation A2058G in one of two 23S rRNA operons. The heterozygosity of these mutants indicates that A2058G confers a dominant or codominant phenotype in this organism. This mutation provides a valuable tool for the genetic manipulation of the 23S rRNA genes of Thermus.Genetic analysis of rRNAs and ribosomal proteins has been an essential tool in elucidating the mechanism of protein synthesis. While numerous ribosomal protein and rRNA mutants have been isolated, none has been examined structurally in atomic detail. This is primarily due to the inadequacy of ribosomes from genetically tractable organisms such as Escherichia coli for forming crystals that diffract X-rays at sufficient resolution. Ribosomes suitable for X-ray crystallographic analysis have thus far been obtained exclusively from thermophiles (4, 23, 33) or halophiles (1). Regrettably, only limited effort has been devoted to studying the ribosomes of these organisms using genetic approaches (see, however, reference 26). Determining the structures of mutant ribosomes of such an organism (an organism producing ribosomes suitable for X-ray crystallographic analysis) should define conformational perturbations that result in specific functional consequences. We have, therefore, initiated a genetic dissection of ribosomes of the extreme thermophile Thermus thermophilus. Despite the promise of Thermus as a genetic system (16, 21), the lack of selectable genetic markers such as transposons or antibiotic resistance mutations makes the mapping of mutations to ribosomal protein and RNA genes problematic. Here we report the isolation of a 23S rRNA mutation conferring resistance to the antibiotics erythromycin and lincomycin. This selectable genetic marker will facilitate the identification, by transformation, of 23S rRNA mutations affecting 50S subunit structure and function.Thermus spp. have an unusual rRNA gene composition of two 16S rRNA genes unlinked from two 23S rRNA-5S rRNAtRNA Gly operons (3, 12, 13). Thus, rRNA mutations are potentially obtainable by direct selection if they exhibit a dominant or codominant phenotype or if gene conversion occurs at a sufficiently high frequency. For these experiments we chose the halotolerant strain T. thermophilus IB-21 (ATCC 43615). Although the strain was originally described as Thermus sp. strain IB-21 (17), the designation T. thermophilus is indicated by DNA-DNA hybridization experiments (32) and our own DNA sequence analysis (S. Gregory, J. H. D. Cate, and A. E. Dahlberg, unpublished results).Isolation of spontaneous erythromycin-resistant mutants. We conducted a search for erythromycin resistance mutations in 23S rRNA which have been isolated for a number of organisms with multiple rRNA operons (reviewed in reference 30). In several instances, including in the cases of Streptomyces ambofaciens (22), Helicobacter pylori (29), and Streptococcus pneumoniae (27), heterozygous mutants indicate that this mutati...

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Isolation and Characterisation of the ylmE Homologue of Thermus thermophilus

et al. 2001

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Screening of a Thermus thermophilus genomic library led to the identification of a homologue of the ylmE gene. ylmE is highly conserved in widely divergent organisms from prokaryotes to mammals, suggesting an important, albeit currently unknown, cellular function. The 633 bp gene has a GC content of 69.2% overall and 90% in the third nucleotide position, while the gene product is predicted to be a soluble cytoplasmic protein of 23,441 Da. It belongs to a family of conserved proteins of unknown function and exhibits amino acid identities ranging from 45% to 28% to the Aquifex aeolicus and Saccharomyces cerevisiae family members, respectively. We speculate that the gene product may be involved in a cellular stress response in T. thermophilus.

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Genes and Genetic Manipulation in Thermus thermophilus

Cited by 3 publications

References 73 publications

Extremophiles in a changing world

Extremophiles in a changing world

Spontaneous Erythromycin Resistance Mutation in a 23S rRNA Gene, rrlA , of the Extreme Thermophile Thermus thermophilus IB-21

Isolation and Characterisation of the ylmE Homologue of Thermus thermophilus

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