A periplasmic cinched protein is required for siderophore secretion and virulence of Mycobacterium tuberculosis - Nature Communications

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A periplasmic cinched protein is required for siderophore secretion and virulence of Mycobacterium tuberculosis - Nature Communications
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A gene in tuberculosis bacteria is found essential for siderophore secretion and virulence NatureComms

reached ~1.0. Afterward, the cells were grown in self-made low-iron 7H9 medium containing the supplements for 5–7 days to deplete intracellular iron sources. The iron-depleted cells were inoculated into 10 mL of low-iron 7H9 medium at an initial ODof 0.01 with different iron sources: 10 μM hemin; 10 μM hemin plus 100 nM Fe- mycobactin; 10 μM hemin plus 1 μM Fe-carboxymycobactin, respectively. The ODSiderophore toxicity assays.

One hundred microliter volumes of each cell suspension were distributed to all wells receiving cells. Plates were incubated for 5–6 days at 37 °C with 230 rpm shaking on a microplate mixer. Forty μL of alamarBlue working solution was added and the plates were incubated at 37 °C with 230 rpm for 8 h. The fluorescence of each well was analyzed using a Biotek Synergy HT plate reader with a 530 nm excitation and a 590 nm emission filter.

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