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[Preprint]. 2023 May 8:2023.05.07.539761. [Version 1] doi: 10.1101/2023.05.07.539761

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Figure 5. — The induction of alx expression in alkaline pH and its dependence on [Mn²⁺]. (A) β-galactosidase activity (Miller units) as a reporter of alx translation (P_alx-PRE-alx’-locZ, pRA54) was measured in mid-log phase grown cultures of Δalx::Kan derivative (RAS31) bearing an empty vector (pHYD5001) or pHYD5001 expressing Alx from a P_trc promoter (pRA27). The cultures were cultivated in LBK media with pHs 6.8 and 8.4, supplemented with appropriate concentration of ampicillin, MnCh and IPTG. (B) β-galactosidase activity (Miller units) as a reporter of alx translation (P_alx-PRE-alx’-locZ, pRA54) was measured in mid-log phase grown cultures of alx⁺ strain (MC4100) in LBK media with pHs 6.8 and 8.4. (C) β-galactosidase activity (Miller units) as a reporter of alx translation (P_alx-PRE-alx’-lacZ, pRA54) was measured in mid-log phase grown cultures of Δalx::Kan derivative (RAS31) in LBK media with pHs 6.8 and 8.4 supplemented with 10 mM sodium citrate. (D) β-galactosidase activity (Miller units) as a reporter of alx translation (P_alx-PRE-alx’-lacZ, pRA54) was measured in mid-log phase grown cultures of Δalx mutant (RAS40) and its ΔmntH::Kan derivative (RAS93) in LBK media with pH 6.8 and 8.4. The error shown is standard deviation of three repeats of the experiment.