2021
DOI: 10.1038/s41467-021-24971-3
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A fructose/H+ symporter controlled by a LacI-type regulator promotes survival of pandemic Vibrio cholerae in seawater

Abstract: The bacterium Vibrio cholerae can colonize the human intestine and cause cholera, but spends much of its life cycle in seawater. The pathogen must adapt to substantial environmental changes when moving between seawater and the human intestine, including different availability of carbon sources such as fructose. Here, we use in vitro experiments as well as mouse intestinal colonization assays to study the mechanisms used by pandemic V. cholerae to adapt to these environmental changes. We show that a LacI-type r… Show more

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Cited by 7 publications
(3 citation statements)
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“…To determine the growth curve of each strain, overnight cultures were washed with PBS 3 times and diluted (1:1000) in LB without antibiotics. A 200 μL aliquot was added to a 96-well flat-bottom microplate, and 200 μL LB medium was used as negative control and incubated at 37 °C with shaking at 180 rpm for 24 h, as previously described ( Liu et al., 2021 ). The absorbance at 600 nm was recorded.…”
Section: Methodsmentioning
confidence: 99%
“…To determine the growth curve of each strain, overnight cultures were washed with PBS 3 times and diluted (1:1000) in LB without antibiotics. A 200 μL aliquot was added to a 96-well flat-bottom microplate, and 200 μL LB medium was used as negative control and incubated at 37 °C with shaking at 180 rpm for 24 h, as previously described ( Liu et al., 2021 ). The absorbance at 600 nm was recorded.…”
Section: Methodsmentioning
confidence: 99%
“…PCR fragments encompassing the target regions were amplified with and without 6-FAM-labeled primers and gel-purified (Sparkjade; AE0101). In each case, 5 ng of each DNA probe was incubated with increasing concentrations of proteins in binding buffer (5 mM HEPES (pH 7.9), 40 mM KCl, 1 mM dithiothreitol (DTT), 0.1 mM EDTA and 5% glycerol, with or without 30 mM acetyl phosphate) 56 . For CarR gel mobility shift assays, 30 mM acetyl phosphate was added to the binding buffer for generating phosphorylated, active CarR.…”
Section: Methodsmentioning
confidence: 99%
“… 52 For aerobic condition, bacteria were grown at 37°C with shaking at 180 rpm. 53 For anaerobic condition, bacteria were grown at 37°C in an anaerobic incubator (YQX-II, Shanghai, China) and oxygen-free nitrogen was used as the carrier gas. 54 Antibiotics were used as following concentrations: polymyxin B, 40 μg/mL; ampicillin, 50 μg/mL; chloramphenicol, 25 μg/mL.…”
Section: Methodsmentioning
confidence: 99%