m9 mucus broth  (Millipore)


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    Name:
    M9 Broth
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    Catalog Number:
    63011
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    Applications:
    M9 Broth is used as a microbiological growth media.
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    Structured Review

    Millipore m9 mucus broth
    Mucus-specific candidate fitness genes are important for growth on the LCFA oleate. Four genes were chosen from the Tn-seq mucus-specific candidate fitness gene list, disrupted, and competed against the WT during microaerobic growth in M9-glucose broth (A) or <t>M9-mucus</t> broth (B). WT strain F11 and each mutant strain were mixed 1:1 and grown microaerobically for 24 h in the indicated medium and then subcultured 1:100 and grown for another 24 h. The titers in the cultures were determined at both the 24- and 48-h time points. The log 10 CI was calculated by dividing the mutant titers by the WT titers (Δ/WT). *, the CI was significantly different from 0 by the one-sample t test ( P ≤ 0.05). (B) Bacteria were grown and titers were determined as described in the legend to panel A, except that the bacteria were grown in M9-mucus medium. *, the CI was significantly different from 0 by the one-sample t test ( P ≤ 0.05). (C) Mutant strains that exhibited defects in M9-mucus broth were complemented with plasmids that expressed the specified target genes under the control of their native promoters (F11 Δ fadL was complemented with pCWR37, the Δ fbp mutant was complemented with pCWR38, and the Δ glpG mutant was complemented with pCWR39). The graph shows the results from competitive assays between the WT strain carrying the empty vector pCWR40 and mutant strains that were either complemented or transformed with pCWR40. P values were determined by an unpaired Student's t test. **, P ≤ 0.01; ****, P ≤ 0.0001. The bars in panels A to C indicate mean values ± SEMs from three independent experiments performed in triplicate. (D and E) The Δ fadL , Δ fbp , and Δ glpG mutants carrying either an empty vector control or a complementation plasmid, as described in the legend to panel C, were streaked onto agar plates containing oleate (D) or glucose (E) as the sole carbon source.

    https://www.bioz.com/result/m9 mucus broth/product/Millipore
    Average 99 stars, based on 1 article reviews
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    1) Product Images from "The Rhomboid Protease GlpG Promotes the Persistence of Extraintestinal Pathogenic Escherichia coli within the Gut"

    Article Title: The Rhomboid Protease GlpG Promotes the Persistence of Extraintestinal Pathogenic Escherichia coli within the Gut

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00866-16

    Mucus-specific candidate fitness genes are important for growth on the LCFA oleate. Four genes were chosen from the Tn-seq mucus-specific candidate fitness gene list, disrupted, and competed against the WT during microaerobic growth in M9-glucose broth (A) or M9-mucus broth (B). WT strain F11 and each mutant strain were mixed 1:1 and grown microaerobically for 24 h in the indicated medium and then subcultured 1:100 and grown for another 24 h. The titers in the cultures were determined at both the 24- and 48-h time points. The log 10 CI was calculated by dividing the mutant titers by the WT titers (Δ/WT). *, the CI was significantly different from 0 by the one-sample t test ( P ≤ 0.05). (B) Bacteria were grown and titers were determined as described in the legend to panel A, except that the bacteria were grown in M9-mucus medium. *, the CI was significantly different from 0 by the one-sample t test ( P ≤ 0.05). (C) Mutant strains that exhibited defects in M9-mucus broth were complemented with plasmids that expressed the specified target genes under the control of their native promoters (F11 Δ fadL was complemented with pCWR37, the Δ fbp mutant was complemented with pCWR38, and the Δ glpG mutant was complemented with pCWR39). The graph shows the results from competitive assays between the WT strain carrying the empty vector pCWR40 and mutant strains that were either complemented or transformed with pCWR40. P values were determined by an unpaired Student's t test. **, P ≤ 0.01; ****, P ≤ 0.0001. The bars in panels A to C indicate mean values ± SEMs from three independent experiments performed in triplicate. (D and E) The Δ fadL , Δ fbp , and Δ glpG mutants carrying either an empty vector control or a complementation plasmid, as described in the legend to panel C, were streaked onto agar plates containing oleate (D) or glucose (E) as the sole carbon source.
    Figure Legend Snippet: Mucus-specific candidate fitness genes are important for growth on the LCFA oleate. Four genes were chosen from the Tn-seq mucus-specific candidate fitness gene list, disrupted, and competed against the WT during microaerobic growth in M9-glucose broth (A) or M9-mucus broth (B). WT strain F11 and each mutant strain were mixed 1:1 and grown microaerobically for 24 h in the indicated medium and then subcultured 1:100 and grown for another 24 h. The titers in the cultures were determined at both the 24- and 48-h time points. The log 10 CI was calculated by dividing the mutant titers by the WT titers (Δ/WT). *, the CI was significantly different from 0 by the one-sample t test ( P ≤ 0.05). (B) Bacteria were grown and titers were determined as described in the legend to panel A, except that the bacteria were grown in M9-mucus medium. *, the CI was significantly different from 0 by the one-sample t test ( P ≤ 0.05). (C) Mutant strains that exhibited defects in M9-mucus broth were complemented with plasmids that expressed the specified target genes under the control of their native promoters (F11 Δ fadL was complemented with pCWR37, the Δ fbp mutant was complemented with pCWR38, and the Δ glpG mutant was complemented with pCWR39). The graph shows the results from competitive assays between the WT strain carrying the empty vector pCWR40 and mutant strains that were either complemented or transformed with pCWR40. P values were determined by an unpaired Student's t test. **, P ≤ 0.01; ****, P ≤ 0.0001. The bars in panels A to C indicate mean values ± SEMs from three independent experiments performed in triplicate. (D and E) The Δ fadL , Δ fbp , and Δ glpG mutants carrying either an empty vector control or a complementation plasmid, as described in the legend to panel C, were streaked onto agar plates containing oleate (D) or glucose (E) as the sole carbon source.

    Techniques Used: Mutagenesis, Plasmid Preparation, Transformation Assay

    The Δ glpG mutant exhibits a defect in the downstream gene glpR . (A) A schematic of the glpEGR operon, with promoter locations indicated by black arrows. Below the operon are listed the fitness scores (FS; log 2 scale) and P values for each gene in mucus broth determined by Tn-seq. (B) Each glp mutant carrying the empty vector control (pCWR40), a plasmid expressing glpEGR (pCWR39), or a plasmid expressing glpEG (pCWR50) was grown in competition with WT strain F11 carrying the control plasmid. The bacteria were grown in M9-mucus broth for 24 h microaerobically and then subcultured into fresh medium and grown for an additional 24 h. After 48 h of growth, the titer of each culture was determined to enumerate WT and mutant levels. P values were determined by an unpaired Student's t test. *, P ≤ 0.05; ****, P ≤ 0.0001; ns, not significant. Bars indicate mean values ± SEMs from three independent experiments performed in triplicate. (C and D) The WT and Δ glpEGR mutant strains carrying either the empty vector control or the glpEGR or glpEG expression construct, as indicated, were grown on minimal medium agar containing oleate (C) or glucose (D) as the sole source of carbon.
    Figure Legend Snippet: The Δ glpG mutant exhibits a defect in the downstream gene glpR . (A) A schematic of the glpEGR operon, with promoter locations indicated by black arrows. Below the operon are listed the fitness scores (FS; log 2 scale) and P values for each gene in mucus broth determined by Tn-seq. (B) Each glp mutant carrying the empty vector control (pCWR40), a plasmid expressing glpEGR (pCWR39), or a plasmid expressing glpEG (pCWR50) was grown in competition with WT strain F11 carrying the control plasmid. The bacteria were grown in M9-mucus broth for 24 h microaerobically and then subcultured into fresh medium and grown for an additional 24 h. After 48 h of growth, the titer of each culture was determined to enumerate WT and mutant levels. P values were determined by an unpaired Student's t test. *, P ≤ 0.05; ****, P ≤ 0.0001; ns, not significant. Bars indicate mean values ± SEMs from three independent experiments performed in triplicate. (C and D) The WT and Δ glpEGR mutant strains carrying either the empty vector control or the glpEGR or glpEG expression construct, as indicated, were grown on minimal medium agar containing oleate (C) or glucose (D) as the sole source of carbon.

    Techniques Used: Mutagenesis, Plasmid Preparation, Expressing, Construct

    Related Articles

    In Vivo:

    Article Title: Identification of a Membrane-Bound Transcriptional Regulator That Links Chitin and Natural Competence in Vibrio cholerae
    Article Snippet: Libraries were generated by in vivo transposon mutagenesis using a Tn10 vector (pDL1098), which conferred spectinomycin resistance. .. The libraries for this study were prepared in a complex medium we have termed “spectacular broth” (SB): 1× M9 salts (Sigma-Aldrich), 1.2% peptone (Bacto), 2.4% yeast extract (Fisher Scientific), 5 mM MgSO4 , 200 µM CaCl2 , and 0.4% glucose.

    Selection:

    Article Title: Escherichia coli attachment to model particulates: The effects of bacterial cell characteristics and particulate properties
    Article Snippet: Paragraph title: E . coli strain selection and preparation ... One sediment strain (No. 122) did not grow sufficiently in M9 minimal broth (Sigma-Aldrich, St. Louis, MO), so further analyses were based on 44 sediment strains and 33 water strains.

    Article Title: Nutrient-responsive regulation determines biodiversity in a colicin-mediated bacterial community
    Article Snippet: TurboRFP and mCherry labelled colicin E2 resistant non-producer strains were obtained by plating an overnight culture of sensitive strains (BN171 and BN1073) on colicin E2 selective plates to obtain strains BN1175 and BN1075 (LB plates containing 100 μ L crude colicin E2 obtained and filter sterilized from an overnight culture of producer cells were used for selection). .. M9 broth (Sigma-Aldrich) and Lysogeny broth (LB, Sigma-Aldrich) were prepared according to the manufacturer’s instructions.

    In Vitro:

    Article Title: Behavioral heterogeneity in quorum sensing can stabilize social cooperation in microbial populations
    Article Snippet: .. In vitro evolution assay WT PAO1 cells from a single colony were first inoculated into LB broth and cultured at 37 °C with shaking (220 rpm) for 12 h. Bacterial cells were then harvested, and about 1.0–2.0 × 107 cells were separately inoculated into 1.0 ml of QSM [M9–0.5% (w /v ) casein (Sigma)], blank M9 broth, LB broth, 4-fold diluted LB broth (1/4-LB), M9 broth supplemented with 0.5% or 0.1% casamino acids (CAA, Sigma) and cultured at 37 °C for 24 h with shaking (220 rpm). ..

    Article Title: The Rhomboid Protease GlpG Promotes the Persistence of Extraintestinal Pathogenic Escherichia coli within the Gut
    Article Snippet: For in vitro growth experiments in M9-glucose or M9-mucus broth, the above-described recipe was used, except that Casamino Acids were excluded. .. In addition, for the M9-mucus broth, 0.5% porcine gastric mucus (catalog number M1778; Sigma) was used in place of glucose.

    Mutagenesis:

    Article Title: Identification of a Membrane-Bound Transcriptional Regulator That Links Chitin and Natural Competence in Vibrio cholerae
    Article Snippet: Libraries were generated by in vivo transposon mutagenesis using a Tn10 vector (pDL1098), which conferred spectinomycin resistance. .. The libraries for this study were prepared in a complex medium we have termed “spectacular broth” (SB): 1× M9 salts (Sigma-Aldrich), 1.2% peptone (Bacto), 2.4% yeast extract (Fisher Scientific), 5 mM MgSO4 , 200 µM CaCl2 , and 0.4% glucose.

    Article Title: Behavioral heterogeneity in quorum sensing can stabilize social cooperation in microbial populations
    Article Snippet: In vitro evolution assay WT PAO1 cells from a single colony were first inoculated into LB broth and cultured at 37 °C with shaking (220 rpm) for 12 h. Bacterial cells were then harvested, and about 1.0–2.0 × 107 cells were separately inoculated into 1.0 ml of QSM [M9–0.5% (w /v ) casein (Sigma)], blank M9 broth, LB broth, 4-fold diluted LB broth (1/4-LB), M9 broth supplemented with 0.5% or 0.1% casamino acids (CAA, Sigma) and cultured at 37 °C for 24 h with shaking (220 rpm). .. Moreover, 1.0–2.0 × 107 CFUs of rhlI mutant (signal deficient strain in the rhl QS system) cells were also repeatedly subcultured in 1.0 ml of QSM with/without exogenous C4HSL (20 μM) signal molecule (Sigma).

    Isolation:

    Article Title: Identification of a Membrane-Bound Transcriptional Regulator That Links Chitin and Natural Competence in Vibrio cholerae
    Article Snippet: The libraries for this study were prepared in a complex medium we have termed “spectacular broth” (SB): 1× M9 salts (Sigma-Aldrich), 1.2% peptone (Bacto), 2.4% yeast extract (Fisher Scientific), 5 mM MgSO4 , 200 µM CaCl2 , and 0.4% glucose. .. Genomic DNA from this library was isolated and served as the “input.” Then, this library was either diluted back into SB to allow for 13 generations of growth or placed onto chitin flakes for transformation assays.

    Article Title: Escherichia coli attachment to model particulates: The effects of bacterial cell characteristics and particulate properties
    Article Snippet: After isolation, the strains were inoculated in Luria-Bertani liquid media (BD Biosciences; San Jose, CA), grown to the stationary phase, and stored at -80°C in 15% glycerol. .. One sediment strain (No. 122) did not grow sufficiently in M9 minimal broth (Sigma-Aldrich, St. Louis, MO), so further analyses were based on 44 sediment strains and 33 water strains.

    Cell Culture:

    Article Title: Behavioral heterogeneity in quorum sensing can stabilize social cooperation in microbial populations
    Article Snippet: .. In vitro evolution assay WT PAO1 cells from a single colony were first inoculated into LB broth and cultured at 37 °C with shaking (220 rpm) for 12 h. Bacterial cells were then harvested, and about 1.0–2.0 × 107 cells were separately inoculated into 1.0 ml of QSM [M9–0.5% (w /v ) casein (Sigma)], blank M9 broth, LB broth, 4-fold diluted LB broth (1/4-LB), M9 broth supplemented with 0.5% or 0.1% casamino acids (CAA, Sigma) and cultured at 37 °C for 24 h with shaking (220 rpm). ..

    Incubation:

    Article Title: Symmetry and scale orient Min protein patterns in shaped bacterial sculptures
    Article Snippet: Cell-sculpting technique An overnight E. coli bacterial culture incubated at 30°C was back diluted to OD600 = 0.02 in fresh M9 medium supplemented with 4 μM A22 (Merk Chemicals Ltd.), and incubated for 3.5 hours at 30°C. .. The droplet was then immediately covered by a 4.8% agarose pad supplemented with M9 broth, 0.4% glucose, 0.25% protein hydrolysate amicase, 4 μM A22 and 25 μg/ml cephalexin (Sigma-Aldrich).

    Article Title: Occurrence of Virulence-Associated Properties in Enterobacter cloacae
    Article Snippet: Briefly, E. coli indicator strain LG1522 cells ( ) were grown to 5 × 107 bacteria/ml in 3 ml of M9 broth supplemented with 0.4% glucose, 50 μg of thiamine per ml, and 200 μM α,α′-dipyridyl (all from Sigma Chemical Company, St. Louis, Mo.). .. Testing strains grown in the same broth medium were spotted onto the indicator strain, and plates were incubated for 48 h at 37°C.

    Article Title: Characteristics of Dogs with Biofilm‐Forming Escherichia Coli Urinary Tract Infections. Characteristics of dogs with biofilm‐forming Escherichia coli urinary tract infections
    Article Snippet: After enrichment, isolates were adjusted to 0.5 McFarland (approximately 1.5 × 108 CFU/mL) and 10 µL were transferred into 90 µL of M9 broth (Ingredients from Sigma Aldrich, St. Louis, Missouri) in a 96‐well flat‐bottom polystyrene tissue culture treated plate (MBEC 96‐well biofilm inoculator, Innovotech, Edmonton, AB, Canada). .. After overnight incubation, the bacterial suspension was removed and the wells were washed with 200 µL of phosphate buffered saline (PBS) 3 times to remove any remaining, unattached bacteria.

    Article Title: Escherichia coli attachment to model particulates: The effects of bacterial cell characteristics and particulate properties
    Article Snippet: One sediment strain (No. 122) did not grow sufficiently in M9 minimal broth (Sigma-Aldrich, St. Louis, MO), so further analyses were based on 44 sediment strains and 33 water strains. .. E . coli isolates were inoculated in 10 mL of M9 broth and incubated to early stationary phase (1.0 < OD600 < 1.5).

    Article Title: Nutrient-responsive regulation determines biodiversity in a colicin-mediated bacterial community
    Article Snippet: After overnight incubation at 37 °C on colicin E2 selective plates, resistant colonies were selected. .. M9 broth (Sigma-Aldrich) and Lysogeny broth (LB, Sigma-Aldrich) were prepared according to the manufacturer’s instructions.

    Microscopy:

    Article Title: Symmetry and scale orient Min protein patterns in shaped bacterial sculptures
    Article Snippet: The droplet was then immediately covered by a 4.8% agarose pad supplemented with M9 broth, 0.4% glucose, 0.25% protein hydrolysate amicase, 4 μM A22 and 25 μg/ml cephalexin (Sigma-Aldrich). .. The base-plate was subsequently mounted onto the microscope stage and incubated at 26 °C during the course of the experiments.

    Sequencing:

    Article Title: Escherichia coli attachment to model particulates: The effects of bacterial cell characteristics and particulate properties
    Article Snippet: Using primer (GTG)5 with the sequence of 5’-GTGGTGGTGGTGGTG-3’ [ ], we differentiated the 400 E . coli strains by rep-PCR. .. One sediment strain (No. 122) did not grow sufficiently in M9 minimal broth (Sigma-Aldrich, St. Louis, MO), so further analyses were based on 44 sediment strains and 33 water strains.

    Article Title: Nutrient-responsive regulation determines biodiversity in a colicin-mediated bacterial community
    Article Snippet: Sequencing of the btuB region of all colicin E2-resistant strains revealed the presence of IS2 elements. .. M9 broth (Sigma-Aldrich) and Lysogeny broth (LB, Sigma-Aldrich) were prepared according to the manufacturer’s instructions.

    Generated:

    Article Title: Identification of a Membrane-Bound Transcriptional Regulator That Links Chitin and Natural Competence in Vibrio cholerae
    Article Snippet: Libraries were generated by in vivo transposon mutagenesis using a Tn10 vector (pDL1098), which conferred spectinomycin resistance. .. The libraries for this study were prepared in a complex medium we have termed “spectacular broth” (SB): 1× M9 salts (Sigma-Aldrich), 1.2% peptone (Bacto), 2.4% yeast extract (Fisher Scientific), 5 mM MgSO4 , 200 µM CaCl2 , and 0.4% glucose.

    other:

    Article Title: Tanzawaic Acids, a Chemically Novel Set of Bacterial Conjugation Inhibitors
    Article Snippet: M9 broth (Sigma-Aldrich) were used to resuspend bacteria after mating and perform serial dilutions.

    Article Title: Slow unloading leads to DNA-bound β2-sliding clamp accumulation in live Escherichia coli cells
    Article Snippet: One litre of M9 growth medium contains 10.5 g l−1 of autoclaved M9 broth (Sigma-Aldrich); 0.1 mM of autoclaved CaCl2 (Sigma-Aldrich); 0.1 mM of autoclaved MgSO4 (J.T.Baker); 0.3% of filter-sterilized glycerol (Sigma-Aldrich) as carbon source; 0.1 g l−1 of filter-sterilized five amino acids, namely L -threonine, L -leucine, L -proline, L -histidine and L -arginine (all from Sigma-Aldrich) and 10 μl of 0.5% filter-sterilized thiamine (Sigma-Aldrich).

    Crystal Violet Assay:

    Article Title: Characteristics of Dogs with Biofilm‐Forming Escherichia Coli Urinary Tract Infections. Characteristics of dogs with biofilm‐forming Escherichia coli urinary tract infections
    Article Snippet: Paragraph title: 2.5. Crystal violet assay to assess biofilm‐forming capability ... After enrichment, isolates were adjusted to 0.5 McFarland (approximately 1.5 × 108 CFU/mL) and 10 µL were transferred into 90 µL of M9 broth (Ingredients from Sigma Aldrich, St. Louis, Missouri) in a 96‐well flat‐bottom polystyrene tissue culture treated plate (MBEC 96‐well biofilm inoculator, Innovotech, Edmonton, AB, Canada).

    Knock-Out:

    Article Title: The Rhomboid Protease GlpG Promotes the Persistence of Extraintestinal Pathogenic Escherichia coli within the Gut
    Article Snippet: While creating knockout, knock-in, and complemented strains, bacteria were grown in Luria-Bertani (LB) broth. .. In addition, for the M9-mucus broth, 0.5% porcine gastric mucus (catalog number M1778; Sigma) was used in place of glucose.

    Modification:

    Article Title: The Rhomboid Protease GlpG Promotes the Persistence of Extraintestinal Pathogenic Escherichia coli within the Gut
    Article Snippet: When growing bacteria for mouse inoculation experiments, we utilized a modified M9 medium containing MgSO4 ·7H2 O (1 mM), CaCl2 ·2H2 O (0.1 mM), d -(+)-glucose (0.1%), nicotinic acid (0.00125%), thiamine HCl (0.00165%), Casamino Acids (0.2%), Na2 HPO4 (6 g/liter), KH2 PO4 (3 g/liter), NH4 Cl (1 g/liter), and NaCl (0.5 g/liter), all of which were dissolved in water. .. In addition, for the M9-mucus broth, 0.5% porcine gastric mucus (catalog number M1778; Sigma) was used in place of glucose.

    Staining:

    Article Title: Characteristics of Dogs with Biofilm‐Forming Escherichia Coli Urinary Tract Infections. Characteristics of dogs with biofilm‐forming Escherichia coli urinary tract infections
    Article Snippet: After enrichment, isolates were adjusted to 0.5 McFarland (approximately 1.5 × 108 CFU/mL) and 10 µL were transferred into 90 µL of M9 broth (Ingredients from Sigma Aldrich, St. Louis, Missouri) in a 96‐well flat‐bottom polystyrene tissue culture treated plate (MBEC 96‐well biofilm inoculator, Innovotech, Edmonton, AB, Canada). .. The remaining biofilm was stained with 0.1% crystal violet solution and allowed to sit at room temperature for 15 minutes, after which excess stain was removed by washing the plate 3 times with PBS.

    Transformation Assay:

    Article Title: Identification of a Membrane-Bound Transcriptional Regulator That Links Chitin and Natural Competence in Vibrio cholerae
    Article Snippet: The libraries for this study were prepared in a complex medium we have termed “spectacular broth” (SB): 1× M9 salts (Sigma-Aldrich), 1.2% peptone (Bacto), 2.4% yeast extract (Fisher Scientific), 5 mM MgSO4 , 200 µM CaCl2 , and 0.4% glucose. .. Genomic DNA from this library was isolated and served as the “input.” Then, this library was either diluted back into SB to allow for 13 generations of growth or placed onto chitin flakes for transformation assays.

    Spectrophotometry:

    Article Title: Escherichia coli attachment to model particulates: The effects of bacterial cell characteristics and particulate properties
    Article Snippet: One sediment strain (No. 122) did not grow sufficiently in M9 minimal broth (Sigma-Aldrich, St. Louis, MO), so further analyses were based on 44 sediment strains and 33 water strains. .. Time to early stationary phase of the isolates was determined by absorbance at 600 nm (spectrophotometer, HACH, Loveland, CO).

    Cellular Antioxidant Activity Assay:

    Article Title: Behavioral heterogeneity in quorum sensing can stabilize social cooperation in microbial populations
    Article Snippet: .. In vitro evolution assay WT PAO1 cells from a single colony were first inoculated into LB broth and cultured at 37 °C with shaking (220 rpm) for 12 h. Bacterial cells were then harvested, and about 1.0–2.0 × 107 cells were separately inoculated into 1.0 ml of QSM [M9–0.5% (w /v ) casein (Sigma)], blank M9 broth, LB broth, 4-fold diluted LB broth (1/4-LB), M9 broth supplemented with 0.5% or 0.1% casamino acids (CAA, Sigma) and cultured at 37 °C for 24 h with shaking (220 rpm). ..

    Genetically Modified:

    Article Title: Escherichia coli attachment to model particulates: The effects of bacterial cell characteristics and particulate properties
    Article Snippet: A pathogenic strain was also investigated: ATCCTM 43888, a genetically modified version of E . coli O157:H7, with the genes that produce Shiga-like toxins I and II removed. .. One sediment strain (No. 122) did not grow sufficiently in M9 minimal broth (Sigma-Aldrich, St. Louis, MO), so further analyses were based on 44 sediment strains and 33 water strains.

    Derivative Assay:

    Article Title: Nutrient-responsive regulation determines biodiversity in a colicin-mediated bacterial community
    Article Snippet: CFP and YFP labelled resistant strains (BN1056 and BN1085, respectively) were directly derived from BN1011 by inserting fragments coding for the fluorescent proteins in the lacIZ region. .. M9 broth (Sigma-Aldrich) and Lysogeny broth (LB, Sigma-Aldrich) were prepared according to the manufacturer’s instructions.

    Knock-In:

    Article Title: The Rhomboid Protease GlpG Promotes the Persistence of Extraintestinal Pathogenic Escherichia coli within the Gut
    Article Snippet: While creating knockout, knock-in, and complemented strains, bacteria were grown in Luria-Bertani (LB) broth. .. In addition, for the M9-mucus broth, 0.5% porcine gastric mucus (catalog number M1778; Sigma) was used in place of glucose.

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    Millipore m9 minimal medium
    Relative nitric oxide resistance of wild-type and mutant S . Typhimurium strains. Growth of the wild type and the cyaY , yggX , and cyaY yggX mutant strains was monitored in the presence of the NO· donor Sper/NO. Strains were inoculated at a density of 10 6 CFU ml −1 in <t>M9</t> minimal medium plus 0.2% gluconate with (open symbols) or without (closed symbols) 750 μM Sper/NO. Growth was monitored in a Bioscreen C microplate reader with agitation at 37°C. Data shown are means ± 1 standard deviation from at least three independent experiments.
    M9 Minimal Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m9 minimal medium/product/Millipore
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    95
    Millipore m9 casein medium
    Fitness outcomes from single-growth-cycle competitions between different V. harveyi genotypes. In pairwise competitions between different V. harveyi strains, the starting competitor frequency was altered and competition experiments were completed in <t>M9-casein</t> medium for 24 h. Each point corresponds to a single competition outcome, and fit lines consist of regression fits (solid lines) bounded by 95% confidence intervals (dashed lines) for each competition pairing, according to a linear model. For each pairing, the strain whose relative fitness is reported on the y axis and starting percentage on the x axis is underlined.
    M9 Casein Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m9 casein medium/product/Millipore
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    99
    Millipore integrin specific antibodies
    Immobilized ISM promotes EC adhesion through α v β 5 <t>integrin.</t> ( A ) ECs attach to ISM-coated surface in a dose-dependent and saturable manner. ( B ) Similar to VN, ISM supports EC adhesion and spreading, induces integrin clustering and actin stress fiber formation. Cell spreading in the absence or presence of EDTA are shown in phase contrast micrographs (panels A – D , scale bar: 40 μ m). Actin stress fibers (red) and β 5 integrin clustering (green) are shown in panels E–H (scale bar: 20 μ m). ( C ) ISM mediates EC adhesion through α v β 5 integrin. ( D ) Knockdown of β 3 and β 5 integrin suppresses EC adhesion to VN. ( E ) Knockdown of β 5 integrin suppresses EC adhesion to ISM
    Integrin Specific Antibodies, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Relative nitric oxide resistance of wild-type and mutant S . Typhimurium strains. Growth of the wild type and the cyaY , yggX , and cyaY yggX mutant strains was monitored in the presence of the NO· donor Sper/NO. Strains were inoculated at a density of 10 6 CFU ml −1 in M9 minimal medium plus 0.2% gluconate with (open symbols) or without (closed symbols) 750 μM Sper/NO. Growth was monitored in a Bioscreen C microplate reader with agitation at 37°C. Data shown are means ± 1 standard deviation from at least three independent experiments.

    Journal: Infection and Immunity

    Article Title: Distinct Roles of the Salmonella enterica Serovar Typhimurium CyaY and YggX Proteins in the Biosynthesis and Repair of Iron-Sulfur Clusters

    doi: 10.1128/IAI.01022-13

    Figure Lengend Snippet: Relative nitric oxide resistance of wild-type and mutant S . Typhimurium strains. Growth of the wild type and the cyaY , yggX , and cyaY yggX mutant strains was monitored in the presence of the NO· donor Sper/NO. Strains were inoculated at a density of 10 6 CFU ml −1 in M9 minimal medium plus 0.2% gluconate with (open symbols) or without (closed symbols) 750 μM Sper/NO. Growth was monitored in a Bioscreen C microplate reader with agitation at 37°C. Data shown are means ± 1 standard deviation from at least three independent experiments.

    Article Snippet: To measure growth in the presence of nitric oxide (NO· ), overnight cultures were diluted to a final OD600 of ∼0.001 in M9 minimal medium with 0.2% gluconate containing 750 μM spermine NONOate (Sper/NO; EMD Chemicals, Billerica, MA).

    Techniques: Mutagenesis, Standard Deviation

    Fitness outcomes from single-growth-cycle competitions between different V. harveyi genotypes. In pairwise competitions between different V. harveyi strains, the starting competitor frequency was altered and competition experiments were completed in M9-casein medium for 24 h. Each point corresponds to a single competition outcome, and fit lines consist of regression fits (solid lines) bounded by 95% confidence intervals (dashed lines) for each competition pairing, according to a linear model. For each pairing, the strain whose relative fitness is reported on the y axis and starting percentage on the x axis is underlined.

    Journal: Applied and Environmental Microbiology

    Article Title: Bacterial Quorum Sensing Stabilizes Cooperation by Optimizing Growth Strategies

    doi: 10.1128/AEM.01945-16

    Figure Lengend Snippet: Fitness outcomes from single-growth-cycle competitions between different V. harveyi genotypes. In pairwise competitions between different V. harveyi strains, the starting competitor frequency was altered and competition experiments were completed in M9-casein medium for 24 h. Each point corresponds to a single competition outcome, and fit lines consist of regression fits (solid lines) bounded by 95% confidence intervals (dashed lines) for each competition pairing, according to a linear model. For each pairing, the strain whose relative fitness is reported on the y axis and starting percentage on the x axis is underlined.

    Article Snippet: Supernatants were obtained from cultures grown in M9-casein medium for 24 h. Cells were removed by first pelleting at 10,000 × g and then filtered with 0.22-μm filters (Millipore).

    Techniques:

    Invasion of the Δ luxOU strain but not the WT strain by the Δ luxR defector. Populations contained one of the two cooperator strains, i.e., the WT strain or the Δ luxOU strain, and the Δ luxR defector at a 99:1 starting mixture, or vice versa. Cultures were grown for 24 h in M9-casein medium, diluted 1,000-fold, and repeated over multiple growth cycles. (A) Defector frequency (determined as nonluminescent colonies) plotted versus generations of growth. (B) Population productivity (measured as CFU per milliliter) plotted versus generations of growth. Error bars, 95% confidence intervals ( n = 3 biological replicates per treatment).

    Journal: Applied and Environmental Microbiology

    Article Title: Bacterial Quorum Sensing Stabilizes Cooperation by Optimizing Growth Strategies

    doi: 10.1128/AEM.01945-16

    Figure Lengend Snippet: Invasion of the Δ luxOU strain but not the WT strain by the Δ luxR defector. Populations contained one of the two cooperator strains, i.e., the WT strain or the Δ luxOU strain, and the Δ luxR defector at a 99:1 starting mixture, or vice versa. Cultures were grown for 24 h in M9-casein medium, diluted 1,000-fold, and repeated over multiple growth cycles. (A) Defector frequency (determined as nonluminescent colonies) plotted versus generations of growth. (B) Population productivity (measured as CFU per milliliter) plotted versus generations of growth. Error bars, 95% confidence intervals ( n = 3 biological replicates per treatment).

    Article Snippet: Supernatants were obtained from cultures grown in M9-casein medium for 24 h. Cells were removed by first pelleting at 10,000 × g and then filtered with 0.22-μm filters (Millipore).

    Techniques:

    QS regulation of extracellular protease activity in V. harveyi . (A) Protease levels for cooperator and defector genotypes grown in M9-casein medium, plotted versus growth (measured as CFU per milliliter) ( n = 3). Error bars, standard errors of the mean. (B) Effects of supernatant supplementation. Cultures of the Δ luxR strain were supplemented with supernatants from the strains indicated, at the indicated concentrations ( n = 4). Error bars, 95% confidence intervals. ***, P

    Journal: Applied and Environmental Microbiology

    Article Title: Bacterial Quorum Sensing Stabilizes Cooperation by Optimizing Growth Strategies

    doi: 10.1128/AEM.01945-16

    Figure Lengend Snippet: QS regulation of extracellular protease activity in V. harveyi . (A) Protease levels for cooperator and defector genotypes grown in M9-casein medium, plotted versus growth (measured as CFU per milliliter) ( n = 3). Error bars, standard errors of the mean. (B) Effects of supernatant supplementation. Cultures of the Δ luxR strain were supplemented with supernatants from the strains indicated, at the indicated concentrations ( n = 4). Error bars, 95% confidence intervals. ***, P

    Article Snippet: Supernatants were obtained from cultures grown in M9-casein medium for 24 h. Cells were removed by first pelleting at 10,000 × g and then filtered with 0.22-μm filters (Millipore).

    Techniques: Activity Assay

    QS requirement for maximal growth of V. harveyi in M9-casein medium. Six strains of V. harveyi were grown for 24 h in M9 medium with different sole carbon sources and were enumerated through viable cell counting (CFU per milliliter). (A) Bar graph showing genotype productivity. Bars, productivity of a given genotype at the culmination of a single growth cycle (24 h) ( n = 4 separate biological replicates); error bars, 95% confidence intervals for the mean estimates. WT, wild-type strain; SN, signal-negative strain; D47E, luxO D47E strain. Differences that were statistically significant, compared with the WT strain in the given medium type, are reported. *, P

    Journal: Applied and Environmental Microbiology

    Article Title: Bacterial Quorum Sensing Stabilizes Cooperation by Optimizing Growth Strategies

    doi: 10.1128/AEM.01945-16

    Figure Lengend Snippet: QS requirement for maximal growth of V. harveyi in M9-casein medium. Six strains of V. harveyi were grown for 24 h in M9 medium with different sole carbon sources and were enumerated through viable cell counting (CFU per milliliter). (A) Bar graph showing genotype productivity. Bars, productivity of a given genotype at the culmination of a single growth cycle (24 h) ( n = 4 separate biological replicates); error bars, 95% confidence intervals for the mean estimates. WT, wild-type strain; SN, signal-negative strain; D47E, luxO D47E strain. Differences that were statistically significant, compared with the WT strain in the given medium type, are reported. *, P

    Article Snippet: Supernatants were obtained from cultures grown in M9-casein medium for 24 h. Cells were removed by first pelleting at 10,000 × g and then filtered with 0.22-μm filters (Millipore).

    Techniques: Cell Counting

    Immobilized ISM promotes EC adhesion through α v β 5 integrin. ( A ) ECs attach to ISM-coated surface in a dose-dependent and saturable manner. ( B ) Similar to VN, ISM supports EC adhesion and spreading, induces integrin clustering and actin stress fiber formation. Cell spreading in the absence or presence of EDTA are shown in phase contrast micrographs (panels A – D , scale bar: 40 μ m). Actin stress fibers (red) and β 5 integrin clustering (green) are shown in panels E–H (scale bar: 20 μ m). ( C ) ISM mediates EC adhesion through α v β 5 integrin. ( D ) Knockdown of β 3 and β 5 integrin suppresses EC adhesion to VN. ( E ) Knockdown of β 5 integrin suppresses EC adhesion to ISM

    Journal: Cell Death & Disease

    Article Title: Isthmin exerts pro-survival and death-promoting effect on endothelial cells through alphavbeta5 integrin depending on its physical state

    doi: 10.1038/cddis.2011.37

    Figure Lengend Snippet: Immobilized ISM promotes EC adhesion through α v β 5 integrin. ( A ) ECs attach to ISM-coated surface in a dose-dependent and saturable manner. ( B ) Similar to VN, ISM supports EC adhesion and spreading, induces integrin clustering and actin stress fiber formation. Cell spreading in the absence or presence of EDTA are shown in phase contrast micrographs (panels A – D , scale bar: 40 μ m). Actin stress fibers (red) and β 5 integrin clustering (green) are shown in panels E–H (scale bar: 20 μ m). ( C ) ISM mediates EC adhesion through α v β 5 integrin. ( D ) Knockdown of β 3 and β 5 integrin suppresses EC adhesion to VN. ( E ) Knockdown of β 5 integrin suppresses EC adhesion to ISM

    Article Snippet: In some experiments, ECs were pre-incubated for 30 min with integrin-specific antibodies (α v antibody (clone M9), β 1 antibody (clone P4C10), β 3 antibody (clone 25E11), β 5 antibody (clone E19), α vβ 3 antibody (clone LM609), α vβ 5 antibody (clone P1F6)) (Millipore, Billerica, MA, USA) or control IgG prior to their addition to the wells.

    Techniques:

    ISM induces EC apoptosis through α v β 5 integrin. ( a ) Knockdown of β 5, but not β 3 integrin, suppressed ISM-induced apoptosis. ( b ) Knockdown of β 5 integrin rescued ISM-suppressed proliferation. ( c ) More free α v β 5 integrins are available in ECs growing on FN-coated surface. VN, vitronectin; ISM, isthmin; FN, fibronectin. ( d ) ISM induces more extensive apoptosis in ECs grown on FN-coated surface

    Journal: Cell Death & Disease

    Article Title: Isthmin exerts pro-survival and death-promoting effect on endothelial cells through alphavbeta5 integrin depending on its physical state

    doi: 10.1038/cddis.2011.37

    Figure Lengend Snippet: ISM induces EC apoptosis through α v β 5 integrin. ( a ) Knockdown of β 5, but not β 3 integrin, suppressed ISM-induced apoptosis. ( b ) Knockdown of β 5 integrin rescued ISM-suppressed proliferation. ( c ) More free α v β 5 integrins are available in ECs growing on FN-coated surface. VN, vitronectin; ISM, isthmin; FN, fibronectin. ( d ) ISM induces more extensive apoptosis in ECs grown on FN-coated surface

    Article Snippet: In some experiments, ECs were pre-incubated for 30 min with integrin-specific antibodies (α v antibody (clone M9), β 1 antibody (clone P4C10), β 3 antibody (clone 25E11), β 5 antibody (clone E19), α vβ 3 antibody (clone LM609), α vβ 5 antibody (clone P1F6)) (Millipore, Billerica, MA, USA) or control IgG prior to their addition to the wells.

    Techniques:

    Identification of the α v β 5 integrin-binding site in ISM. ( a ) Schematic diagrams showing the truncated ISM protein fragments produced as recombinant proteins in E. coli . Numbers refer to the amino acid position in the full-length secreted ISM protein. The positon of KD316AA mutation is indicated by the arrowhead. The black triangle at the right end of each truncated protein represent His-tag. ( b ) EC adhesion to various ISM protein fragments compared with 0.2% gelatin. 3% BSA was used as the negative control. ( c ) KD316AA mutation abolished the anti-tube formation activity of ISM

    Journal: Cell Death & Disease

    Article Title: Isthmin exerts pro-survival and death-promoting effect on endothelial cells through alphavbeta5 integrin depending on its physical state

    doi: 10.1038/cddis.2011.37

    Figure Lengend Snippet: Identification of the α v β 5 integrin-binding site in ISM. ( a ) Schematic diagrams showing the truncated ISM protein fragments produced as recombinant proteins in E. coli . Numbers refer to the amino acid position in the full-length secreted ISM protein. The positon of KD316AA mutation is indicated by the arrowhead. The black triangle at the right end of each truncated protein represent His-tag. ( b ) EC adhesion to various ISM protein fragments compared with 0.2% gelatin. 3% BSA was used as the negative control. ( c ) KD316AA mutation abolished the anti-tube formation activity of ISM

    Article Snippet: In some experiments, ECs were pre-incubated for 30 min with integrin-specific antibodies (α v antibody (clone M9), β 1 antibody (clone P4C10), β 3 antibody (clone 25E11), β 5 antibody (clone E19), α vβ 3 antibody (clone LM609), α vβ 5 antibody (clone P1F6)) (Millipore, Billerica, MA, USA) or control IgG prior to their addition to the wells.

    Techniques: Binding Assay, Produced, Recombinant, Mutagenesis, Negative Control, Activity Assay

    Schematic illustration of the dual functions of ISM. ( a ) Immobilized ISM serves as integrin agonist to mediate cell survival/migration by activating FAK. ( b ) Soluble ISM induces cell apoptosis through IMD (direct activation of caspase-8). ‘?' indicates the unresolved question if additional molecules are involved in the integrin–caspase-8 complex formation

    Journal: Cell Death & Disease

    Article Title: Isthmin exerts pro-survival and death-promoting effect on endothelial cells through alphavbeta5 integrin depending on its physical state

    doi: 10.1038/cddis.2011.37

    Figure Lengend Snippet: Schematic illustration of the dual functions of ISM. ( a ) Immobilized ISM serves as integrin agonist to mediate cell survival/migration by activating FAK. ( b ) Soluble ISM induces cell apoptosis through IMD (direct activation of caspase-8). ‘?' indicates the unresolved question if additional molecules are involved in the integrin–caspase-8 complex formation

    Article Snippet: In some experiments, ECs were pre-incubated for 30 min with integrin-specific antibodies (α v antibody (clone M9), β 1 antibody (clone P4C10), β 3 antibody (clone 25E11), β 5 antibody (clone E19), α vβ 3 antibody (clone LM609), α vβ 5 antibody (clone P1F6)) (Millipore, Billerica, MA, USA) or control IgG prior to their addition to the wells.

    Techniques: Migration, Radial Immuno Diffusion, Activation Assay

    ISM induces apoptosis through IMD. ( A ) ISM induced apoptosis through caspase-8-mediated extrinsic pathway, but not caspase-9-mediated intrinsic pathway. ( B ) Kinetics of ISM-induced apoptosis. Activated caspase-8 can first be detected 4 h after ISM treatment. ( C ) Knockdown of β 5-integrin subunit by siRNA suppressed caspase-8 and caspase-3 activation, as well as cleavage of PARP. ( D ) ISM induces caspase-8 association with α v β 5 integrin and activation. ( E ) Soluble ISM induces α v β 5-integrin clustering. Co-localization of β 5 integrin (red) with ISM (green) is indicated by yellow signals. DAPI (blue) stains the nucleus. Scale bar: 20 μ m

    Journal: Cell Death & Disease

    Article Title: Isthmin exerts pro-survival and death-promoting effect on endothelial cells through alphavbeta5 integrin depending on its physical state

    doi: 10.1038/cddis.2011.37

    Figure Lengend Snippet: ISM induces apoptosis through IMD. ( A ) ISM induced apoptosis through caspase-8-mediated extrinsic pathway, but not caspase-9-mediated intrinsic pathway. ( B ) Kinetics of ISM-induced apoptosis. Activated caspase-8 can first be detected 4 h after ISM treatment. ( C ) Knockdown of β 5-integrin subunit by siRNA suppressed caspase-8 and caspase-3 activation, as well as cleavage of PARP. ( D ) ISM induces caspase-8 association with α v β 5 integrin and activation. ( E ) Soluble ISM induces α v β 5-integrin clustering. Co-localization of β 5 integrin (red) with ISM (green) is indicated by yellow signals. DAPI (blue) stains the nucleus. Scale bar: 20 μ m

    Article Snippet: In some experiments, ECs were pre-incubated for 30 min with integrin-specific antibodies (α v antibody (clone M9), β 1 antibody (clone P4C10), β 3 antibody (clone 25E11), β 5 antibody (clone E19), α vβ 3 antibody (clone LM609), α vβ 5 antibody (clone P1F6)) (Millipore, Billerica, MA, USA) or control IgG prior to their addition to the wells.

    Techniques: Radial Immuno Diffusion, Activation Assay

    ISM inhibits in vitro angiogenesis through α v β 5 integrin. ( a ) Transient transfection of β 3 and β 5 siRNA effectively knocked down their expression in ECs. Same amount of corresponding scrambled siRNA are transfected as control. ( b ) Only knockdown of β 5-integrin expression abolished the anti-tube formation activity of ISM

    Journal: Cell Death & Disease

    Article Title: Isthmin exerts pro-survival and death-promoting effect on endothelial cells through alphavbeta5 integrin depending on its physical state

    doi: 10.1038/cddis.2011.37

    Figure Lengend Snippet: ISM inhibits in vitro angiogenesis through α v β 5 integrin. ( a ) Transient transfection of β 3 and β 5 siRNA effectively knocked down their expression in ECs. Same amount of corresponding scrambled siRNA are transfected as control. ( b ) Only knockdown of β 5-integrin expression abolished the anti-tube formation activity of ISM

    Article Snippet: In some experiments, ECs were pre-incubated for 30 min with integrin-specific antibodies (α v antibody (clone M9), β 1 antibody (clone P4C10), β 3 antibody (clone 25E11), β 5 antibody (clone E19), α vβ 3 antibody (clone LM609), α vβ 5 antibody (clone P1F6)) (Millipore, Billerica, MA, USA) or control IgG prior to their addition to the wells.

    Techniques: In Vitro, Transfection, Expressing, Activity Assay

    Immobilized ISM activates FAK and promotes EC haptotactic migration. ( a ) Immobilized ISM activates FAK Y397 phosphorylation in similar fashion as immobilized VN. S, cells in suspension. ( b ) ISM dose-dependently promotes EC haptotactic migration with similar potency as FN. ( c ) ISM promotes EC haptotactic migration through α v β 5 integrin

    Journal: Cell Death & Disease

    Article Title: Isthmin exerts pro-survival and death-promoting effect on endothelial cells through alphavbeta5 integrin depending on its physical state

    doi: 10.1038/cddis.2011.37

    Figure Lengend Snippet: Immobilized ISM activates FAK and promotes EC haptotactic migration. ( a ) Immobilized ISM activates FAK Y397 phosphorylation in similar fashion as immobilized VN. S, cells in suspension. ( b ) ISM dose-dependently promotes EC haptotactic migration with similar potency as FN. ( c ) ISM promotes EC haptotactic migration through α v β 5 integrin

    Article Snippet: In some experiments, ECs were pre-incubated for 30 min with integrin-specific antibodies (α v antibody (clone M9), β 1 antibody (clone P4C10), β 3 antibody (clone 25E11), β 5 antibody (clone E19), α vβ 3 antibody (clone LM609), α vβ 5 antibody (clone P1F6)) (Millipore, Billerica, MA, USA) or control IgG prior to their addition to the wells.

    Techniques: Migration