Archaeal genetics — the third way

Allers, Thorsten; Mevarech, Moshe

doi:10.1038/nrg1504

Cited by 224 publications

(215 citation statements)

References 151 publications

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“…Tightly regulated gene expression systems are among the most needed genetic tools in research with archaea (Allers andMevarech 2005, Rother et al 2005). Existing expression systems in methanoarchaea are based on fusions of the gene of interest to a catabolic promoter involved, e.g., in methanol, acetate utilization or assimilation of nitrogenous compounds (Apolinario et al 2005, Lei et al 2005.…”

Section: Discussionmentioning

confidence: 99%

New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species

Guss

Rother

Zhang

et al. 2008

Archaea

117

179

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Summary A highly efficient method for chromosomal integration of cloned DNA into Methanosarcina spp. was developed utilizing the site-specific recombination system from the Streptomyces phage φC31. Host strains expressing the φC31 integrase gene and carrying an appropriate recombination site can be transformed with non-replicating plasmids carrying the complementary recombination site at efficiencies similar to those obtained with self-replicating vectors. We have also constructed a series of hybrid promoters that combine the highly expressed M. barkeri PmcrB promoter with binding sites for the tetracycline-responsive, bacterial TetR protein. These promoters are tightly regulated by the presence or absence of tetracycline in strains that express the tetR gene. The hybrid promoters can be used in genetic experiments to test gene essentiality by placing a gene of interest under their control. Thus, growth of strains with tetR-regulated essential genes becomes tetracycline-dependent. A series of plasmid vectors that utilize the site-specific recombination system for construction of reporter gene fusions and for tetracycline regulated expression of cloned genes are reported. These vectors were used to test the efficiency of translation at a variety of start codons. Fusions using an ATG start site were the most active, whereas those using GTG and TTG were approximately one half or one fourth as active, respectively. The CTG fusion was 95% less active than the ATG fusion.

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Section: Discussionmentioning

confidence: 99%

New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species

Guss

Rother

Zhang

et al. 2008

Archaea

117

179

View full text Add to dashboard Cite

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“…The growing number of archaeal genome sequences revealed an increasing number of putative transcriptional regulators, some of which have been characterized at the molecular level (Allers and Mevarech, 2005;Bell, 2005).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the TrmB‐like protein, PF0124, a TGM‐recognizing global transcriptional regulator of the hyperthermophilic archaeon Pyrococcus furiosus

Lee

Surma

Seitz

et al. 2007

Molecular Microbiology

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“…Interestingly, although there are some known archaea that growth in mesophilic conditions, most of them are extremophiles. Two main phylogenetic groups can be found inside Archaea: Euryarchaeota and Crenarchaeota (Allers & Mevarech, 2005). Also, recently based in environmental samples, two more groups called Korarchaeota and Nanoarchaeota has been proposed (Allers & Mevarech, 2005).…”

Section: Glucose Degradation In the Members Of The Archaea Domainmentioning

confidence: 99%