Transcriptional regulatory networks (TRNs) in bacteria are crucial for elucidating the mechanisms that regulate gene expression and cellular responses to environmental stimuli. These networks delineate the interactions between transcription factors (TFs) and their target genes, thereby uncovering the regulatory processes that modulate gene expression under varying environmental conditions. Analyzing TRNs offers valuable insights into bacterial adaptation, stress responses, and metabolic optimization from an evolutionary standpoint.
Additionally, understanding TRNs can drive the development of novel antimicrobial therapies and the engineering of microbial strains for biofuel and bioproduct production. This protocol integrates advanced data analysis pipelines, including ChEAP, DEOCSU, and DESeq2, to analyze omics datasets that encompass genome-wide TF binding sites and transcriptome profiles derived from ChIP-exo and RNA-seq experiments. This approach minimizes both the time required and the risk of bias, making it accessible to non-expert users. Key steps in the protocol include preprocessing and peak calling from ChIP-exo data, differential expression analysis of RNA-seq data, and motif and regulon analysis. This method offers a comprehensive and efficient framework for TRN reconstruction across various bacterial strains, enhancing both the accuracy and reliability of the analysis while providing valuable insights for basic and applied research.
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These amino acids are quite different in their amino acid properties (polar, non-polar, hydrophobicity), but one commonality is that they are all relatively small. Larger amino acid substitutions such as histidine, methionine, and tyrosine were made site-directedly and showed to have no detectable DNA binding, further supporting the requirement of small amino acids at CRP position 183.
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the PE_PGRS45 protein. We found that overexpression of this cell wall-associated protein enhanced bacterial viability under
stress in vitro and cell survival in macrophages. MS_PE_PGRS45 decreased the secretion of pro-inflammatory cytokines such
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genome engineering method to efficiently obtain engineered phages in a model bacterial pathogen, Pseudomonas aeruginosa.
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the recombinant generation based on the Red recombination system of the coliphage λ (λRed). The producer (PD) cells of
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assays. This study aimed to optimize the PPIA by determining
a suitable reaction terminator and an optimal methanol
concentration in the sample. The most suitable reaction time
was 90 min, with the corresponding methanol concentration
in the sample being 15% or less. When p-nitrophenyl phosphate
(pNPP) was used as a substrate, copper chloride solution
was suitably used as a reaction terminator, and when 4-
methylumbelliferyl phosphate (MUP) was used, a glycine buffer
not only increased the measurement sensitivity of the reaction
product but also terminated the enzymatic reaction.
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respectively, the limit of quantitation for MC-leucine/
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pNPP was used as a substrate. The proposed method facilitated
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additional pretreatments, such as concentration or purification;
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process, rapid analysis time, and high-resolution phenotypic
data. Advanced statistical techniques (e.g., denoising
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diversity by processing the scatter data obtained from
flow cytometry. These phenotypic diversities were well correlated
with taxonomic-based diversity computed using nextgeneration
16S RNA gene sequencing. The protocol provided
in this paper should be a useful guide for a fast and reliable
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mediumTM; SC and SF, Spores formed in an agricultural byproduct
medium with 10 mM CaCl2 and with 10 mM FeSO4,
respectively). Stronger UV resistance was recorded for SF
with 1.8–2.3-fold greater survival than SC and SD under UV
treatment. The three spore types showed similar heat resistances
at 80°C, but survival rates of SC and SD were much
higher (~1,000 times) than those of SF at 90°C. However,
germination capacity of SF was 20% higher than those of
SD and SC on Luria-Bertani agar plates for 24 h. SF germinated
more rapidly in a liquid medium with high NaCl concentrations
than SC and SD, but became slower under alkaline
conditions. Raman spectroscopy was used to analyze the
heterogeneities in the three types of vegetative cells and their
spores under different nutritional conditions. Exponentially
grown-each vegetative cells had different overall Raman peak
values. Raman peaks of SC, SD, and SF also showed differences
in adenine and amide III compositions and nucleic acid
contents. Our data along with Raman spectroscopy provided
the evidence that spores formed under under different growth
conditions possess very different cellular components, which
affected their survival and germination rates.
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and can be found either in a culturable or a viable
but nonculturable (VBNC) state. Despite widespread concerns
as to how to define the occurrence and dynamics of
Vibrio populations by culture-independent approaches, further
physiological research and relevant biotechnological
developments will require the isolation and cultivation of the
microbes from various environments. The present work provides
data and perspectives on our understanding of culturable
Vibrio community structure and diversity in the Beibu
Gulf. Finally, we isolated 1,037 strains of Vibrio from 45 samples
and identified 18 different species. Vibrio alginolyticus,
V. cyclitrophicus, V. tasmaniensis, V. brasiliensis, and V. splendidus
were the dominant species that had regional distribution
characteristics. The correlation between the quantitative
distribution and community structure of culturable Vibrio and
environmental factors varied with the Vibrio species and geographical
locations. Among them, salinity, nitrogen, and phosphorus
were the main factors affecting the diversity of culturable
Vibrio. These results help to fill a knowledge gap on
Vibrio diversity and provide data for predicting and controlling
pathogenic Vibrio outbreaks in the Beibu Gulf.
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Although bacteria have diverse membrane proteins, the function
of many of them remains unknown or uncertain even
in Escherichia coli. In this study, to investigate the function
of hypothetical membrane proteins, genome-wide analysis
of phenotypes of hypothetical membrane proteins was performed
under various envelope stresses. Several genes responsible
for adaptation to envelope stresses were identified.
Among them, deletion of YhcB, a conserved inner membrane
protein of unknown function, caused high sensitivities to various
envelope stresses and increased membrane permeability,
and caused growth defect under normal growth conditions.
Furthermore, yhcB deletion resulted in morphological
aberration, such as branched shape, and cell division defects,
such as filamentous growth and the generation of chromosome-
less cells. The analysis of antibiotic susceptibility
showed that the yhcB mutant was highly susceptible to various
anti-folate antibiotics. Notably, all phenotypes of the yhcB
mutant were completely or significantly restored by YhcB
without the transmembrane domain, indicating that the localization
of YhcB on the inner membrane is dispensable for its
function. Taken together, our results demonstrate that YhcB
is involved in cell morphology and cell division in a membrane
localization-independent manner.
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We employed a stepwise selection model for investigating the
dynamics of antibiotic-resistant variants in Escherichia coli
K-12 treated with increasing concentrations of ciprofloxacin
(CIP). Firstly, we used Sanger sequencing to screen the variations
in the fluoquinolone target genes, then, employed Illumina
NGS sequencing for amplicons targeted regions with
variations. The results demonstrated that variations G81C in
gyrA and K276N and K277L in parC are standing resistance
variations (SRVs), while S83A and S83L in gyrA and G78C
in parC were emerging resistance variations (ERVs). The variants
containing SRVs and/or ERVs were selected successively
based on their sensitivities to CIP. Variant strain 1, containing
substitution G81C in gyrA, was immediately selected
following ciprofloxacin exposure, with obvious increases in
the parC SRV, and parC and gyrA ERV allele frequencies.
Variant strain 2, containing the SRVs, then dominated the
population following a 20× increase in ciprofloxacin concentration,
with other associated allele frequencies also elevated.
Variant strains 3 and 4, containing ERVs in gyrA and parC,
respectively, were then selected at 40× and 160× antibiotic
concentrations. Two variants, strains 5 and 6, generated in
the selection procedure, were lost because of higher fitness
costs or a lower level of resistance compared with variants 3
and 4. For the second induction, all variations/indels were
already present as SRVs and selected out step by step at different
passages. Whatever the first induction or second induction,
our results confirmed the soft selective sweep hypothesis
and provided critical information for guiding clinical
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on bacteria remain unknown. RNA-seq based transcriptome
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sulfur/taurine transporters were highly expressed as countermeasures
against the phytotoxins. In addition, our findings
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Antibiotic resistance is a global concern in public health.
Antibiotic-resistant clones can spread nationally, internationally,
and globally. This review considers representative
antibiotic-resistant Gram-negative bacterial clones–CTX-M-
15-producing ST131 in Escherichia coli, extended-spectrum
β-lactamase-producing ST11 and KPC-producing ST258 in
Klebsiella pneumoniae, IMP-6-producing, carbapenem-resistant
ST235 in Pseudomonas aeruginosa, and OXA-23-
producing global clone 2 in Acinetobacter baumannii–that
have disseminated worldwide, including in Korea. The findings
highlight the urgency for systematic monitoring and
international cooperation to suppress the emergence and
propagation of antibiotic resistance.
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Many studies have shown that hydrogen sulfide (H2S) is both
detrimental and beneficial to animals and plants, whereas its
effect on bacteria is not fully understood. Here, we report that
H2S, released by sodium hydrosulfide (NaHS), significantly
inhibits the growth of Escherichia coli in a dose-dependent
manner. Further studies have shown that H2S treatment stimulates
the production of reactive oxygen species (ROS) and
decreases glutathione (GSH) levels in E. coli, resulting in lipid
peroxidation and DNA damage. H2S also inhibits the antioxidative
enzyme activities of superoxide dismutase (SOD),
catalase (CAT) and glutathione reductase (GR) and induces
the response of the SoxRS and OxyR regulons in E. coli. Moreover,
pretreatment with the antioxidant ascorbic acid (AsA)
could effectively prevent H2S-induced toxicity in E. coli. Taken
together, our results indicate that H2S exhibits an antibacterial
effect on E. coli through oxidative damage and suggest
a possible application for H2S in water and food processing.
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