E. coli K‐12 strains, plasmids, and promoter fragments used in this study are listed in Table
Lennox broth
Manufactured by Merck Group
Sourced in United Kingdom
Lennox broth is a microbiological medium used for the cultivation and growth of various bacteria. It provides the necessary nutrients and growth conditions for bacterial cultures.
Automatically generated - may contain errors
Lab products found in correlation
Peptone, by Thermo Fisher Scientific (1 mentions)
Yeast extract, by Thermo Fisher Scientific (1 mentions)
Lysogeny broth, by Merck Group (1 mentions)
M9 minimal salts media, by Merck Group (1 mentions)
Lennox broth, by Thermo Fisher Scientific (1 mentions)
Antimicrobial disks, by Thermo Fisher Scientific (1 mentions)
Iso sensitest broth, by Thermo Fisher Scientific (1 mentions)
3 protocols using lennox broth
1
E. coli Strain and Media Preparation
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E. coli K‐12 strains, plasmids, and promoter fragments used in this study are listed in TableS1 and oligonucleotide primers are in Table S2 . Strains were grown in lysogeny broth (LB) (Sigma), Lennox broth (2% (w/v) peptone (Oxoid), 1% (w/v) yeast extract (Oxoid) and 170 mM NaCl), minimal salts medium (Squire et al., 2009 (link)) (minimal salts with 0.4% glycerol, 10% Lennox broth, 20 mM fumarate), or M9 minimal salts media (Sigma), supplemented with 0.36% glucose, 2 mM MgSO4, 0.1 mM CaCl2, all with appropriate antibiotic supplements (ampicillin 100 µg/ml and tetracycline 15 µg/ml).
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E. coli K‐12 strains, plasmids, and promoter fragments used in this study are listed in Table
2
Bacterial Growth and Antibiotic Evaluation
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Strains used in this study are listed in Table 7 . Bacterial strains were grown overnight at 37°C in Lennox broth (Sigma-Aldrich, UK) or Iso-Sensitest broth (Oxoid, UK). All chemicals and antibiotics were supplied by Sigma-Aldrich, UK. Antimicrobial disks were supplied by Oxoid, UK.
3
Protein Expression in E. coli Cultures
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All protein expressions took place at 37 °C in LB media (Lennox broth, Sigma Aldrich, St. Louis, MO) supplemented with either 100 μg/ml ampicillin or 100 μg/ml ampicillin and 25 μg/ml chloramphenicol for strains that carry an additional chloramphenicol selectable vector, namely pLysS, pGro7, RosettaGami, and SoluBL21-pLysS. Strains were cultivated overnight at 37 °C in 5 ml of LB media placed in 15 ml conical tubes at an angle of ∼55° with 225 rpm shaking. One milliliter of overnight cultures was used to inoculate 50 ml expression cultures placed in 250 ml Erlenmeyer flasks with 225 rpm shaking. Recombinant protein production was induced with 1 mM isopropyl-β-ᴅ-thiogalactoside (IPTG; Sigma Aldrich, St. Louis, MO) when the cultures reached an OD600 of 0.6–0.8, typically 2–3 h after inoculation. For experiments where titer and plasmid maintenance were quantified, expression cultures were harvested 4 h after induction. For experiments that generated 10.25-h growth curves, cells were put through the same procedure as above, but cultures were not harvested until 10.25 h after inoculation.
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