The strain H. volcanii H26 was obtained from Thorsten Allers (Nottingham, United Kingdom), it is a pyrE deletion strain lacking the plasmid pHV2. The deletion of the dhfr (dihydrofolate reductase) gene HVO_1279 has been described previously (Maurer et al., 2018 (link)). The deletion strains of genes encoding translation initiation factors have been generated in a previous study (Gäbel et al., 2013 (link)). Multi cycle PCRs were used to confirm that all mutants were still homozygous. The sequences of oligonucleotides are listed in Supplementary Table S1 . In a few cases the mutants could not be regrown from permanent cultures, therefore, they were regenerated as described (Gäbel et al., 2013 (link)) using the oligonucleotides listed in Supplementary Table S1 . All overproduction strains have been generated in this study (see below).
Haloferax volcanii strains were grown in complex medium with 50μg/ml uracil as previously described (Dambeck and Soppa, 2008 (link)). The cultures were grown in Erlenmeyer flasks at 42°C with shaking at 250rpm. Growth was either measured spectroscopically at 600nm or cells were counted using a Neubauer counting chamber.
The E. coli strain XL1-blue MRF’ (Agilent Technologies, Waldbronn, Germany) was used for cloning. It was grown in complex SOB medium under standard conditions (Hanahan, 1983 (link)).
Haloferax volcanii strains were grown in complex medium with 50μg/ml uracil as previously described (Dambeck and Soppa, 2008 (link)). The cultures were grown in Erlenmeyer flasks at 42°C with shaking at 250rpm. Growth was either measured spectroscopically at 600nm or cells were counted using a Neubauer counting chamber.
The E. coli strain XL1-blue MRF’ (Agilent Technologies, Waldbronn, Germany) was used for cloning. It was grown in complex SOB medium under standard conditions (Hanahan, 1983 (link)).
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