7h9 medium Search Results


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  • 99
    Thermo Fisher middlebrook 7h9 medium
    Disruption of pknG or removal of the phosphorylation motif of garA caused a nutrient-dependent growth defect in M . smegmatis . ( A ) All strains grew at the same rate on standard <t>Middlebrook</t> <t>7H9</t> medium. ( B ) Δ garA Ms grew slower than wild type on minimal Sauton’s medium containing 20 mM propionate, 20 mM NH 4 Cl plus 0.05% tyloxapol, and this growth defect could be fully complemented by GarA lacking phosphorylation sites (truncated “trunc.” garA ). ( C + D ) Δ pknG Ms grew slower than wild type and formed clumps (inset photo) on medium containing glutamate as sole carbon ( C ) or nitrogen source ( D ) (minimal Sauton’s with either 30 mM glutamate plus tyloxapol, or 1% glycerol, 10 mM glutamate plus tyloxapol). Data plotted are the mean and standard deviation of at least 3 independent experiments. ( E ) Δ pknG Ms formed clumps when glutamate was the sole carbon or nitrogen source. The photograph shows a microplate from growth curve ( D ) imaged at 60 hours. Growth of Δ garA Ms complemented with phosphoablative GarA (EAAS) was equivalent to that of Δ pknG Ms complemented with truncated GarA in all tested conditions so only the dataset for truncated GarA is shown for clarity.
    Middlebrook 7h9 Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore 7h9 broth medium
    Disruption of pknG or removal of the phosphorylation motif of garA caused a nutrient-dependent growth defect in M . smegmatis . ( A ) All strains grew at the same rate on standard <t>Middlebrook</t> <t>7H9</t> medium. ( B ) Δ garA Ms grew slower than wild type on minimal Sauton’s medium containing 20 mM propionate, 20 mM NH 4 Cl plus 0.05% tyloxapol, and this growth defect could be fully complemented by GarA lacking phosphorylation sites (truncated “trunc.” garA ). ( C + D ) Δ pknG Ms grew slower than wild type and formed clumps (inset photo) on medium containing glutamate as sole carbon ( C ) or nitrogen source ( D ) (minimal Sauton’s with either 30 mM glutamate plus tyloxapol, or 1% glycerol, 10 mM glutamate plus tyloxapol). Data plotted are the mean and standard deviation of at least 3 independent experiments. ( E ) Δ pknG Ms formed clumps when glutamate was the sole carbon or nitrogen source. The photograph shows a microplate from growth curve ( D ) imaged at 60 hours. Growth of Δ garA Ms complemented with phosphoablative GarA (EAAS) was equivalent to that of Δ pknG Ms complemented with truncated GarA in all tested conditions so only the dataset for truncated GarA is shown for clarity.
    7h9 Broth Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Difco 7h9 medium
    Macroscopic morphology of M. smegmatis mc 2 155 strain spreading on the surface of a motility agar plate. mc 2 155 was grown in 7H10, and a single colony was transferred with a toothpick to the center of a 0.3% agar plate containing <t>7H9</t> basal medium without any added carbon source. The plate was sealed with parafilm and incubated at 37°C for 2 weeks.
    7h9 Medium, supplied by Difco, used in various techniques. Bioz Stars score: 95/100, based on 251 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Becton Dickinson 7h9 medium
    In vitro stability of pBP10 in M. smegmatis and Mtb in the absence of antibiotic selection. ( a ) Total number of M. smegmatis -pBP10, accounting for serial dilutions to show ongoing culture expansion. ( b ) Frequency of M. smegmatis -pBP10 plasmid-containing bacteria. ( c ) Frequency of plasmid carriage in M. smegmatis -pBP10 for log-phase cultures versus generations (calculated as (log(OD 600 t / OD 600 ( t − 1)) / log(2)). ( d ) Total number of M. tuberculosis -pBP10. ( e ) Frequency of M. tuberculosis -pBP10 plasmid-containing bacteria. ( f ) Frequency of plasmid carriage in M. tuberculosis -pBP10 for log-phase cultures versus generations. Data (means ± s.d.) are shown for log-phase cultures in <t>7H9,</t> 1:1 7H9 to water, 1:3 7H9 to water, 7H9 without shaking and 7H9 in 2% O 2 . Representative data are shown for two to five experiments performed in triplicate, except for the 2% O 2 experiment, which was performed once.
    7h9 Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 95/100, based on 207 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    HiMedia Laboratories 7h9 medium
    Survival of Erdman strain of M. tb in <t>7H9</t> media. M. tb grown in 7H9 media containing no additives (A) , M. tb grown in 7H9 media containing NAC (B) , M. tb grown in 7H9 media containing INH and INH+ NAC (C) , M. tb grown in 7H9 media containing RIF and RIF + NAC (D) , and M. tb grown in 7H9 media containing EMB and EMB+ NAC (E) . There was a significant increase in bacterial numbers at 15 days when M. tb was grown in 7H9 in the absence of any additives (A) . There was a significant reduction in the bacterial numbers at 15 days when M. tb was grown in 7H9 in the presence of NAC (B) , INH and INH+NAC (C) , RIF and RIF+NAC (D) and EMB and EMB+NAC (E) . Data represent means ±SE from experiments performed in triplicate. * p
    7h9 Medium, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 91/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore 7h9 medium
    MAb 24c5 ELISA analysis and binding to glycogen. (A) Splenic fusion resulted in MAb 24c5, an IgG1 able to recognize M. tuberculosis GC. Each data point represents the average of three measurements. (B) MAb 24c5 binding to dilute concentrations of GC in the M. tuberculosis GC ELISA. Each data point represents the average of three measurements. (C) MAb 24c5 binding to M. tuberculosis Erdman day-20 <t>7H9</t> medium (MTB), cell culture supernatant extract (INTPHSE), GC, AM, lipoarabinomannan (LAM), phosphatidylinositol mannoside (PIM), fast-growing mycobacterial lipomannan (LM), mycolyl-arabinogalactan-peptidoglycan complex (mAGP), and total lipid fraction (TLF). Each bar represents the average of two measurements. (D) Effects of proteinase K on binding of MAb 24c5 to M. tuberculosis Erdman day-20 7H9 medium. (E and F) MAb 24c5 assayed for reactivity to type VIII (slipper limpet) (viii) (E) and type IX (ix) (bovine liver) (F) glycogens. Each data point represents the average of three measurements. Error bars show the standard deviations of the means.
    7h9 Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 163 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    MiddleBrook Pharmaceuticals 7h9 medium
    Survival of Δ cpsA under in vitro stress conditions. ( A ) Growth curve of H37Rv, Δ cpsA , and Δ cpsA::cpsA grown in <t>7H9</t> medium. ( B ) Survival of H37Rv, Δ cpsA , and Δ katG in the presence of 5 mM H 2 O 2 . Samples were plated after 30, 60, 90, 120, and 180 min. No colonies were observed for Δ katG after 30 min (#). *** P = 0.0005. ( C ) Survival of H37Rv and Δ cpsA after the addition of 200 μM diethylenetriamine/nitric oxide (DETA-NO) every 24 h for 3 d. ns, not significant; Student’s t test. ( D ) In vitro growth curves of H37Rv and Δ cpsA in 7H9 medium under different pHs (range 5.0–7.5). Data are representative of two experiments. ( E ) Sensitivity of H37Rv or Δ cpsA in the presence of lysozyme. The indicated concentration of lysozyme was added to 7H11 plates. Data are representative of two independent experiments. * P ≤ 0.05, *** P ≤ 0.0005; Student’s t test. ( F ) Sensitivity of H37Rv and Δ cpsA to SDS. The indicated bacterial numbers were spotted on 7H11 agar plates with SDS (0.005%, 0.01%) or without SDS. Pictures were taken 15 d after spotting. Data represent the results of three independent experiments.
    7h9 Medium, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 95/100, based on 144 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Difco 7h9 liquid medium
    Growth of M. smegmatis in various nitrogen sources. Wild-type SMR5 and glnR deletion strain MH1 were grown in the presence of the indicated substances (10 mM final concentration) as sole nitrogen source. Standard <t>7H9</t> medium was used as positive control; 7H9 lacking any nitrogen source as negative control (7H9-N). (A) Putative nitrogen sources deduced from microarray results, (B) putative nitrogen sources used by closely related species.
    7h9 Liquid Medium, supplied by Difco, used in various techniques. Bioz Stars score: 94/100, based on 236 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Difco middlebrook 7h9 medium
    Kinetics and inducibility of fructose, glucose, and glycerol uptake by M. smegmatis . (A) M. smegmatis mc 2 155 was grown in <t>Middlebrook</t> <t>7H9</t> medium in the presence of 2% glycerol (open circles) or 2% fructose (closed circles). Accumulation
    Middlebrook 7h9 Medium, supplied by Difco, used in various techniques. Bioz Stars score: 95/100, based on 837 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Becton Dickinson middlebrook 7h9 medium
    Cell surface of Msmeg-PE11 is highly hydrophobic. ( a ) Pellicle formation of Msmeg-pVV or Msmeg-PE11 was monitored by growing standing cultures of the strains without shaking in <t>Middlebrook</t> <t>7H9</t> medium in absence of Tween 80 at 37 °C for various time points. ( b ) Biofilm formation was quantified by crystal violet staining for which Msmeg-pVV or Msmeg-PE11 cells were washed, stained with 1% crystal violet and ethanol extract was spectrophotometrically measured at 570 nm. ( c ) Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 medium with (upper panel) or without (lower panel) 0.05% Tween-80 at 37 °C with shaking for 24 h and 48 h. Cultures were then allowed to settle at room temperature for 30 min. ( d ) In another experiment, Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 broth with congo red (100 μg/ml) and 0.05% Tween 80 for 24 h, 48 h and 72 h at 37 °C. Cells were next washed and re-suspended in acetone. Congo red in the supernatant was spectrophotometrically measured at 490 nm. Data are representative of mean ± SD of three different experiments.
    Middlebrook 7h9 Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 97/100, based on 261 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Merck & Co middlebrook 7h9 medium
    Cell surface of Msmeg-PE11 is highly hydrophobic. ( a ) Pellicle formation of Msmeg-pVV or Msmeg-PE11 was monitored by growing standing cultures of the strains without shaking in <t>Middlebrook</t> <t>7H9</t> medium in absence of Tween 80 at 37 °C for various time points. ( b ) Biofilm formation was quantified by crystal violet staining for which Msmeg-pVV or Msmeg-PE11 cells were washed, stained with 1% crystal violet and ethanol extract was spectrophotometrically measured at 570 nm. ( c ) Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 medium with (upper panel) or without (lower panel) 0.05% Tween-80 at 37 °C with shaking for 24 h and 48 h. Cultures were then allowed to settle at room temperature for 30 min. ( d ) In another experiment, Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 broth with congo red (100 μg/ml) and 0.05% Tween 80 for 24 h, 48 h and 72 h at 37 °C. Cells were next washed and re-suspended in acetone. Congo red in the supernatant was spectrophotometrically measured at 490 nm. Data are representative of mean ± SD of three different experiments.
    Middlebrook 7h9 Medium, supplied by Merck & Co, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Difco 7h9 broth medium
    Genetic organization, growth analysis, bacterial survival, and Mtb Δ esxL mutant construction. A , schematic representation of e sxL in the M. tuberculosis H37Rv genome. RAW 264.7 ( B ) and THP-1 ( C ) were infected with M. smegmatis ( Msm ) pSMT3 and recombinant M. smegmatis esxL strains. The cells were lysed, and intracellular survival was determined 1, 8, and 24 h post-infection by a cfu assay. D , in vitro growth curve of the M. smegmatis WT, M. smegmatis pSMT3, and recombinant M. smegmatis esxL was determined by growing bacteria in <t>7H9</t> medium and measuring OD ( O.D 600 nm ) . E , extracellular expression of the esxL transcript was measured by qRT-PCR after growing M. smegmatis esxL in vitro for 4, 12, and 24 h. RNA was isolated at the respective time points. cDNA was synthesized, and the expression of esxL was determined using qRT-PCR. Transcript levels are represented relative to mRNA -fold change of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. F , intracellular expression of esxL transcript was measured by qRT-PCR. RNA was isolated from M. smegmatis esxL -infected macrophages at different time points. cDNA was synthesized, and the expression of esxL was determined using qRT PCR. Transcript levels are represented relative to mRNA level of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. G , schematic representation of construction of Mtb Δ esxL mutant by homologous recombination. The location of primers used for the confirmation of deletion mutant generation is depicted. H , confirmation of Mtb Δ esxL mutant generation. F1 and R2 primers were designed beyond the flanks, whereas R1 and F2 primers anneal to sacB-hyg r cassette. PCR using F1 and R1 is expected to give no product with the M. tuberculosis ( lane 1 ) and ∼1.3 kb with the Mtb Δ esxL ( lane 2 ); F2-R2 primer sets were expected to give no product with M. tuberculosis and ∼1.5 kb in Mtb Δ esxL mutant. Amplification of udgB with gene-specific primers was performed as a control. The experiments were performed in triplicate ( n = 3). Results are shown as mean ± S.D. ( error bars ). *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ns , not significant.
    7h9 Broth Medium, supplied by Difco, used in various techniques. Bioz Stars score: 96/100, based on 95 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher 7h9 liquid medium
    Genetic organization, growth analysis, bacterial survival, and Mtb Δ esxL mutant construction. A , schematic representation of e sxL in the M. tuberculosis H37Rv genome. RAW 264.7 ( B ) and THP-1 ( C ) were infected with M. smegmatis ( Msm ) pSMT3 and recombinant M. smegmatis esxL strains. The cells were lysed, and intracellular survival was determined 1, 8, and 24 h post-infection by a cfu assay. D , in vitro growth curve of the M. smegmatis WT, M. smegmatis pSMT3, and recombinant M. smegmatis esxL was determined by growing bacteria in <t>7H9</t> medium and measuring OD ( O.D 600 nm ) . E , extracellular expression of the esxL transcript was measured by qRT-PCR after growing M. smegmatis esxL in vitro for 4, 12, and 24 h. RNA was isolated at the respective time points. cDNA was synthesized, and the expression of esxL was determined using qRT-PCR. Transcript levels are represented relative to mRNA -fold change of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. F , intracellular expression of esxL transcript was measured by qRT-PCR. RNA was isolated from M. smegmatis esxL -infected macrophages at different time points. cDNA was synthesized, and the expression of esxL was determined using qRT PCR. Transcript levels are represented relative to mRNA level of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. G , schematic representation of construction of Mtb Δ esxL mutant by homologous recombination. The location of primers used for the confirmation of deletion mutant generation is depicted. H , confirmation of Mtb Δ esxL mutant generation. F1 and R2 primers were designed beyond the flanks, whereas R1 and F2 primers anneal to sacB-hyg r cassette. PCR using F1 and R1 is expected to give no product with the M. tuberculosis ( lane 1 ) and ∼1.3 kb with the Mtb Δ esxL ( lane 2 ); F2-R2 primer sets were expected to give no product with M. tuberculosis and ∼1.5 kb in Mtb Δ esxL mutant. Amplification of udgB with gene-specific primers was performed as a control. The experiments were performed in triplicate ( n = 3). Results are shown as mean ± S.D. ( error bars ). *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ns , not significant.
    7h9 Liquid Medium, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Difco 7h9 medium powder
    Genetic organization, growth analysis, bacterial survival, and Mtb Δ esxL mutant construction. A , schematic representation of e sxL in the M. tuberculosis H37Rv genome. RAW 264.7 ( B ) and THP-1 ( C ) were infected with M. smegmatis ( Msm ) pSMT3 and recombinant M. smegmatis esxL strains. The cells were lysed, and intracellular survival was determined 1, 8, and 24 h post-infection by a cfu assay. D , in vitro growth curve of the M. smegmatis WT, M. smegmatis pSMT3, and recombinant M. smegmatis esxL was determined by growing bacteria in <t>7H9</t> medium and measuring OD ( O.D 600 nm ) . E , extracellular expression of the esxL transcript was measured by qRT-PCR after growing M. smegmatis esxL in vitro for 4, 12, and 24 h. RNA was isolated at the respective time points. cDNA was synthesized, and the expression of esxL was determined using qRT-PCR. Transcript levels are represented relative to mRNA -fold change of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. F , intracellular expression of esxL transcript was measured by qRT-PCR. RNA was isolated from M. smegmatis esxL -infected macrophages at different time points. cDNA was synthesized, and the expression of esxL was determined using qRT PCR. Transcript levels are represented relative to mRNA level of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. G , schematic representation of construction of Mtb Δ esxL mutant by homologous recombination. The location of primers used for the confirmation of deletion mutant generation is depicted. H , confirmation of Mtb Δ esxL mutant generation. F1 and R2 primers were designed beyond the flanks, whereas R1 and F2 primers anneal to sacB-hyg r cassette. PCR using F1 and R1 is expected to give no product with the M. tuberculosis ( lane 1 ) and ∼1.3 kb with the Mtb Δ esxL ( lane 2 ); F2-R2 primer sets were expected to give no product with M. tuberculosis and ∼1.5 kb in Mtb Δ esxL mutant. Amplification of udgB with gene-specific primers was performed as a control. The experiments were performed in triplicate ( n = 3). Results are shown as mean ± S.D. ( error bars ). *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ns , not significant.
    7h9 Medium Powder, supplied by Difco, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    BioLife Solutions middlebrook 7h9 medium
    Genetic organization, growth analysis, bacterial survival, and Mtb Δ esxL mutant construction. A , schematic representation of e sxL in the M. tuberculosis H37Rv genome. RAW 264.7 ( B ) and THP-1 ( C ) were infected with M. smegmatis ( Msm ) pSMT3 and recombinant M. smegmatis esxL strains. The cells were lysed, and intracellular survival was determined 1, 8, and 24 h post-infection by a cfu assay. D , in vitro growth curve of the M. smegmatis WT, M. smegmatis pSMT3, and recombinant M. smegmatis esxL was determined by growing bacteria in <t>7H9</t> medium and measuring OD ( O.D 600 nm ) . E , extracellular expression of the esxL transcript was measured by qRT-PCR after growing M. smegmatis esxL in vitro for 4, 12, and 24 h. RNA was isolated at the respective time points. cDNA was synthesized, and the expression of esxL was determined using qRT-PCR. Transcript levels are represented relative to mRNA -fold change of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. F , intracellular expression of esxL transcript was measured by qRT-PCR. RNA was isolated from M. smegmatis esxL -infected macrophages at different time points. cDNA was synthesized, and the expression of esxL was determined using qRT PCR. Transcript levels are represented relative to mRNA level of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. G , schematic representation of construction of Mtb Δ esxL mutant by homologous recombination. The location of primers used for the confirmation of deletion mutant generation is depicted. H , confirmation of Mtb Δ esxL mutant generation. F1 and R2 primers were designed beyond the flanks, whereas R1 and F2 primers anneal to sacB-hyg r cassette. PCR using F1 and R1 is expected to give no product with the M. tuberculosis ( lane 1 ) and ∼1.3 kb with the Mtb Δ esxL ( lane 2 ); F2-R2 primer sets were expected to give no product with M. tuberculosis and ∼1.5 kb in Mtb Δ esxL mutant. Amplification of udgB with gene-specific primers was performed as a control. The experiments were performed in triplicate ( n = 3). Results are shown as mean ± S.D. ( error bars ). *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ns , not significant.
    Middlebrook 7h9 Medium, supplied by BioLife Solutions, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson 7h9 liquid medium
    Growth curves of the WT, ΔMycP 3 mutant and the two complementation strains, ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms under Fe-free <t>7H9</t> ( a ), Fe-free Sauton’s media ( b ). The growth curves were done in triplicate, error bars show standard deviation
    7h9 Liquid Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 111 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Difco 7h9 modified medium
    Anti- Mycobacterium bovis capacity of rHBD3 protein to A549 cells. a M. bovis growth curve after incubation with different concentrations of HBD-3. Negative control is M. bovis with Middle Brook <t>7H9</t> broth. Positive control is M. bovis with streptomycin (1000 U/ml). b Mycobacteria invasion tests. Yellow spots in the figure are Mycobacteria . c Cell apoptosis analysis of A549 cells infected by M. bovis with different treatments. d The data of cell apoptosis and death ratio analysis of A549 cells infected by M. bovis with different treatments. All the experiments were replicated three times and the changes are presented as mean ± SEM. P values
    7h9 Modified Medium, supplied by Difco, used in various techniques. Bioz Stars score: 94/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Becton Dickinson 7h9 oadc medium
    Anti- Mycobacterium bovis capacity of rHBD3 protein to A549 cells. a M. bovis growth curve after incubation with different concentrations of HBD-3. Negative control is M. bovis with Middle Brook <t>7H9</t> broth. Positive control is M. bovis with streptomycin (1000 U/ml). b Mycobacteria invasion tests. Yellow spots in the figure are Mycobacteria . c Cell apoptosis analysis of A549 cells infected by M. bovis with different treatments. d The data of cell apoptosis and death ratio analysis of A549 cells infected by M. bovis with different treatments. All the experiments were replicated three times and the changes are presented as mean ± SEM. P values
    7h9 Oadc Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 95/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    HiMedia Laboratories middelbrook 7h9 medium
    Anti- Mycobacterium bovis capacity of rHBD3 protein to A549 cells. a M. bovis growth curve after incubation with different concentrations of HBD-3. Negative control is M. bovis with Middle Brook <t>7H9</t> broth. Positive control is M. bovis with streptomycin (1000 U/ml). b Mycobacteria invasion tests. Yellow spots in the figure are Mycobacteria . c Cell apoptosis analysis of A549 cells infected by M. bovis with different treatments. d The data of cell apoptosis and death ratio analysis of A549 cells infected by M. bovis with different treatments. All the experiments were replicated three times and the changes are presented as mean ± SEM. P values
    Middelbrook 7h9 Medium, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore 7h9 liquid medium
    Growth of RGM within A. polyphaga trophozoites. M. gilvum (A), M. rhodesiae (B), and M. thermoresistibile (C) were cocultured with the free-living amoeba A. polyphaga (black bars), cultivated in PAS medium (gray bars), and cultivated in <t>7H9</t> complete medium
    7h9 Liquid Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore middlebrook 7h9 medium
    Growth of RGM within A. polyphaga trophozoites. M. gilvum (A), M. rhodesiae (B), and M. thermoresistibile (C) were cocultured with the free-living amoeba A. polyphaga (black bars), cultivated in PAS medium (gray bars), and cultivated in <t>7H9</t> complete medium
    Middlebrook 7h9 Medium, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson 7h9 broth medium
    Growth of RGM within A. polyphaga trophozoites. M. gilvum (A), M. rhodesiae (B), and M. thermoresistibile (C) were cocultured with the free-living amoeba A. polyphaga (black bars), cultivated in PAS medium (gray bars), and cultivated in <t>7H9</t> complete medium
    7h9 Broth Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson 7h9 mycobacterial medium
    Growth of RGM within A. polyphaga trophozoites. M. gilvum (A), M. rhodesiae (B), and M. thermoresistibile (C) were cocultured with the free-living amoeba A. polyphaga (black bars), cultivated in PAS medium (gray bars), and cultivated in <t>7H9</t> complete medium
    7h9 Mycobacterial Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco 7h9 mycobacterial medium
    Growth of RGM within A. polyphaga trophozoites. M. gilvum (A), M. rhodesiae (B), and M. thermoresistibile (C) were cocultured with the free-living amoeba A. polyphaga (black bars), cultivated in PAS medium (gray bars), and cultivated in <t>7H9</t> complete medium
    7h9 Mycobacterial Medium, supplied by Difco, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MiddleBrook Pharmaceuticals 7h9 liquid medium
    Growth of M. tuberculosis strains in liquid and solid media. (A) Wild-type M. tuberculosis H37Rv, Δ cuvA Mt , and complemented Δ cuvA (Δ cuvA Mt /C) strains were grown in <t>7H9-ADC-Tw</t> liquid medium with 0.2% glucose, and the OD 600 was measured daily. (B) Growth of the same strains on agar plates containing MM plus 0.2% glucose or MM plus 0.01% cholesterol. Serial 10-fold dilutions were spotted and incubated at 37°C, and photographs were taken after 1 month of incubation. In both panels, data shown are from one experiment that was repeated at least twice with similar results. Glc, 0.2% glucose; Chol., 0.01% cholesterol.
    7h9 Liquid Medium, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 99/100, based on 616 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MiddleBrook Pharmaceuticals middlebrook 7h9 medium
    FAs of the K10 (Type C) and 2349/06-1 (Type S) isolates of Map that showed statistically significant differences in abundance between the two isolates . BOMAC and MOCL-4 cells were infected with the K10 and 2349/06-1 isolates of Map at MOI of 1:10. At 4 h p.i., the medium containing the extracellular bacteria was collected, centrifuged at 2000 g for 15 min and the FAMEs of the bacterial pellets extracted and analyzed by GC. FAs of the K10 and 2349/06-1 isolates of Map grown in <t>Middlebrook</t> <t>7H9</t> medium that showed statistically significant differences in abundance are also included in the figure. Relative amount of each FA for each experimental condition (7H9 medium or extracellular) is presented as the percentage of the total FAs content. Statistically significant differences are indicated with an asterisk.
    Middlebrook 7h9 Medium, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 99/100, based on 2844 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco 7h9 solid medium
    (A) Resistance of M. smegmatis to CuOOH is dependent in part upon Ohr. Cells were cultured to late stationary phase (17 days) in Middlebrook <t>7H9</t> medium. Cells were then suspended in the same medium at an A 600 of 0.1 and incubated with CuOOH for 2 h at
    7h9 Solid Medium, supplied by Difco, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco 7h9 basal medium
    (A) Resistance of M. smegmatis to CuOOH is dependent in part upon Ohr. Cells were cultured to late stationary phase (17 days) in Middlebrook <t>7H9</t> medium. Cells were then suspended in the same medium at an A 600 of 0.1 and incubated with CuOOH for 2 h at
    7h9 Basal Medium, supplied by Difco, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson l 7h9 medium
    (A) Resistance of M. smegmatis to CuOOH is dependent in part upon Ohr. Cells were cultured to late stationary phase (17 days) in Middlebrook <t>7H9</t> medium. Cells were then suspended in the same medium at an A 600 of 0.1 and incubated with CuOOH for 2 h at
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    Becton Dickinson 2×7h9 broth medium
    (A) Resistance of M. smegmatis to CuOOH is dependent in part upon Ohr. Cells were cultured to late stationary phase (17 days) in Middlebrook <t>7H9</t> medium. Cells were then suspended in the same medium at an A 600 of 0.1 and incubated with CuOOH for 2 h at
    2×7h9 Broth Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Disruption of pknG or removal of the phosphorylation motif of garA caused a nutrient-dependent growth defect in M . smegmatis . ( A ) All strains grew at the same rate on standard Middlebrook 7H9 medium. ( B ) Δ garA Ms grew slower than wild type on minimal Sauton’s medium containing 20 mM propionate, 20 mM NH 4 Cl plus 0.05% tyloxapol, and this growth defect could be fully complemented by GarA lacking phosphorylation sites (truncated “trunc.” garA ). ( C + D ) Δ pknG Ms grew slower than wild type and formed clumps (inset photo) on medium containing glutamate as sole carbon ( C ) or nitrogen source ( D ) (minimal Sauton’s with either 30 mM glutamate plus tyloxapol, or 1% glycerol, 10 mM glutamate plus tyloxapol). Data plotted are the mean and standard deviation of at least 3 independent experiments. ( E ) Δ pknG Ms formed clumps when glutamate was the sole carbon or nitrogen source. The photograph shows a microplate from growth curve ( D ) imaged at 60 hours. Growth of Δ garA Ms complemented with phosphoablative GarA (EAAS) was equivalent to that of Δ pknG Ms complemented with truncated GarA in all tested conditions so only the dataset for truncated GarA is shown for clarity.

    Journal: PLoS Pathogens

    Article Title: PknG senses amino acid availability to control metabolism and virulence of Mycobacterium tuberculosis

    doi: 10.1371/journal.ppat.1006399

    Figure Lengend Snippet: Disruption of pknG or removal of the phosphorylation motif of garA caused a nutrient-dependent growth defect in M . smegmatis . ( A ) All strains grew at the same rate on standard Middlebrook 7H9 medium. ( B ) Δ garA Ms grew slower than wild type on minimal Sauton’s medium containing 20 mM propionate, 20 mM NH 4 Cl plus 0.05% tyloxapol, and this growth defect could be fully complemented by GarA lacking phosphorylation sites (truncated “trunc.” garA ). ( C + D ) Δ pknG Ms grew slower than wild type and formed clumps (inset photo) on medium containing glutamate as sole carbon ( C ) or nitrogen source ( D ) (minimal Sauton’s with either 30 mM glutamate plus tyloxapol, or 1% glycerol, 10 mM glutamate plus tyloxapol). Data plotted are the mean and standard deviation of at least 3 independent experiments. ( E ) Δ pknG Ms formed clumps when glutamate was the sole carbon or nitrogen source. The photograph shows a microplate from growth curve ( D ) imaged at 60 hours. Growth of Δ garA Ms complemented with phosphoablative GarA (EAAS) was equivalent to that of Δ pknG Ms complemented with truncated GarA in all tested conditions so only the dataset for truncated GarA is shown for clarity.

    Article Snippet: Bacterial strains, media, and culture M . tuberculosis H37Rv and M . smegmatis mc2 155 were routinely cultured on Middlebrook 7H10 agar (Oxoid) with 10% ADN (0.5% bovine serum albumin, 0.2% dextrose, 0.085% NaCl) and Middlebrook 7H9 medium (Oxoid) with 10% ADN and 0.05% Tween 80.

    Techniques: Mass Spectrometry, Standard Deviation

    garA is required for growth of M . tuberculosis in vitro , survival in macrophages, and virulence in mice. ( A ) M . tuberculosis lacking garA was unable to grow on standard 7H10 medium unless supplemented with asparagine. Plasmid-borne garA restored the defect, but variants of garA with mutations at threonine 21 in the phosphorylation motif (ETTS) gave only partial complementation. Strains were grown in Middlebrook 7H9 plus 30 mM asparagine then washed and diluted in standard 7H9 and spotted onto standard 7H10 with or without 10 mM asparagine. Photographs are representative of at least 3 independent experiments. ( B ) M . tuberculosis lacking garA (red squares) had a defect in growth and survival in differentiated THP-1 cells compared to parental M . tuberculosis H37Rv (black circles). Re-introduction of GarA (black triangles) or variants of GarA lacking a single phosphorylation site (grey crosses and squares) restored growth but variant GarA lacking both phosphorylation sites (green triangles) did not. Data points show the mean and standard deviation from four replicates and are representative of two independent experiments. ( C ) M . tuberculosis lacking garA was avirulent in mice as it was eliminated from the lungs. BALB/C mice were infected intranasally with 10 5 bacilli and bacterial burden was measured on days 1, 7, 21 and 28. Data points show the bacterial burden in individual animals. The bacterial burden of mice infected with Δ garA Mt (red squares) was significantly lower than those infected with M . tuberculosis H37Rv (black circles), or complemented Δ garA Mt (black triangles) at all time points from day 7 (p

    Journal: PLoS Pathogens

    Article Title: PknG senses amino acid availability to control metabolism and virulence of Mycobacterium tuberculosis

    doi: 10.1371/journal.ppat.1006399

    Figure Lengend Snippet: garA is required for growth of M . tuberculosis in vitro , survival in macrophages, and virulence in mice. ( A ) M . tuberculosis lacking garA was unable to grow on standard 7H10 medium unless supplemented with asparagine. Plasmid-borne garA restored the defect, but variants of garA with mutations at threonine 21 in the phosphorylation motif (ETTS) gave only partial complementation. Strains were grown in Middlebrook 7H9 plus 30 mM asparagine then washed and diluted in standard 7H9 and spotted onto standard 7H10 with or without 10 mM asparagine. Photographs are representative of at least 3 independent experiments. ( B ) M . tuberculosis lacking garA (red squares) had a defect in growth and survival in differentiated THP-1 cells compared to parental M . tuberculosis H37Rv (black circles). Re-introduction of GarA (black triangles) or variants of GarA lacking a single phosphorylation site (grey crosses and squares) restored growth but variant GarA lacking both phosphorylation sites (green triangles) did not. Data points show the mean and standard deviation from four replicates and are representative of two independent experiments. ( C ) M . tuberculosis lacking garA was avirulent in mice as it was eliminated from the lungs. BALB/C mice were infected intranasally with 10 5 bacilli and bacterial burden was measured on days 1, 7, 21 and 28. Data points show the bacterial burden in individual animals. The bacterial burden of mice infected with Δ garA Mt (red squares) was significantly lower than those infected with M . tuberculosis H37Rv (black circles), or complemented Δ garA Mt (black triangles) at all time points from day 7 (p

    Article Snippet: Bacterial strains, media, and culture M . tuberculosis H37Rv and M . smegmatis mc2 155 were routinely cultured on Middlebrook 7H10 agar (Oxoid) with 10% ADN (0.5% bovine serum albumin, 0.2% dextrose, 0.085% NaCl) and Middlebrook 7H9 medium (Oxoid) with 10% ADN and 0.05% Tween 80.

    Techniques: In Vitro, Mouse Assay, Plasmid Preparation, Variant Assay, Standard Deviation, Infection

    Macroscopic morphology of M. smegmatis mc 2 155 strain spreading on the surface of a motility agar plate. mc 2 155 was grown in 7H10, and a single colony was transferred with a toothpick to the center of a 0.3% agar plate containing 7H9 basal medium without any added carbon source. The plate was sealed with parafilm and incubated at 37°C for 2 weeks.

    Journal: Journal of Bacteriology

    Article Title: Sliding Motility in Mycobacteria

    doi:

    Figure Lengend Snippet: Macroscopic morphology of M. smegmatis mc 2 155 strain spreading on the surface of a motility agar plate. mc 2 155 was grown in 7H10, and a single colony was transferred with a toothpick to the center of a 0.3% agar plate containing 7H9 basal medium without any added carbon source. The plate was sealed with parafilm and incubated at 37°C for 2 weeks.

    Article Snippet: M63 or 7H9 medium supplemented as indicated were solidified with 0.3% agar (Difco) or 0.1 to 0.8% ultrapure SeaKem LE agarose (FMC Bioproducts).

    Techniques: Incubation

    Growth accompanies mycobacterial spreading. A 1:100 mix of GFP-labeled (light) and unlabeled (dark) mc 2 155 cells grown as described in Materials and Methods were plated on the surface of 0.3% M63 salts– (A) and 7H9 (with no added carbon source)– (B) agarose plates. Photographs were taken after 2 days of incubation at 37°C. Phase-contrast images showing the continuous spreading halo are on the left, and fluorescent micrographs of the same fields showing the locations of GFP-labeled cells are on the right. Bars, 25 μm.

    Journal: Journal of Bacteriology

    Article Title: Sliding Motility in Mycobacteria

    doi:

    Figure Lengend Snippet: Growth accompanies mycobacterial spreading. A 1:100 mix of GFP-labeled (light) and unlabeled (dark) mc 2 155 cells grown as described in Materials and Methods were plated on the surface of 0.3% M63 salts– (A) and 7H9 (with no added carbon source)– (B) agarose plates. Photographs were taken after 2 days of incubation at 37°C. Phase-contrast images showing the continuous spreading halo are on the left, and fluorescent micrographs of the same fields showing the locations of GFP-labeled cells are on the right. Bars, 25 μm.

    Article Snippet: M63 or 7H9 medium supplemented as indicated were solidified with 0.3% agar (Difco) or 0.1 to 0.8% ultrapure SeaKem LE agarose (FMC Bioproducts).

    Techniques: Labeling, Incubation

    Spreading phenotype of M. avium colony morphology variants 2151-SmD, 2151-SmT, Rg-O, and Rg-4 on 7H9–ADC–0.3% agarose plates. Photographs were taken 3 weeks after inoculation.

    Journal: Journal of Bacteriology

    Article Title: Sliding Motility in Mycobacteria

    doi:

    Figure Lengend Snippet: Spreading phenotype of M. avium colony morphology variants 2151-SmD, 2151-SmT, Rg-O, and Rg-4 on 7H9–ADC–0.3% agarose plates. Photographs were taken 3 weeks after inoculation.

    Article Snippet: M63 or 7H9 medium supplemented as indicated were solidified with 0.3% agar (Difco) or 0.1 to 0.8% ultrapure SeaKem LE agarose (FMC Bioproducts).

    Techniques:

    Pattern formation in a spreading Sm-1 colony. A 25-μl aliquot of a saturated Sm-1 culture was plated onto a 7H9–ADC–0.3% agarose plate. (A) Pictures of the spreading colony taken 1, 2, and 3 days after inoculation. (B and C) Electron micrographs of cells taken at day 3 from the transparent periphery (B) and opaque interior (C) of a spreading colony. Cells were negatively stained with 2% phosphotungstic acid. Bar, 1 μm. Arrows mark the structures discussed in the text.

    Journal: Journal of Bacteriology

    Article Title: Sliding Motility in Mycobacteria

    doi:

    Figure Lengend Snippet: Pattern formation in a spreading Sm-1 colony. A 25-μl aliquot of a saturated Sm-1 culture was plated onto a 7H9–ADC–0.3% agarose plate. (A) Pictures of the spreading colony taken 1, 2, and 3 days after inoculation. (B and C) Electron micrographs of cells taken at day 3 from the transparent periphery (B) and opaque interior (C) of a spreading colony. Cells were negatively stained with 2% phosphotungstic acid. Bar, 1 μm. Arrows mark the structures discussed in the text.

    Article Snippet: M63 or 7H9 medium supplemented as indicated were solidified with 0.3% agar (Difco) or 0.1 to 0.8% ultrapure SeaKem LE agarose (FMC Bioproducts).

    Techniques: Staining

    CtpC Is Involved in Zinc Detoxification and Contributes to the Intracellular Survival of M. tuberculosis (A) Differential sensitivity of M. tuberculosis wild-type and the ctpC null mutant to free zinc. M. tuberculosis wild-type (GC1237), a ctpC null mutant ( ctpC ::Kan R ), or the cosmid-complemented strain (I437) was allowed to grow in 7H9-ADC medium containing 0.1 mM ZnSO 4 or without zinc supplementation (Control). Bacterial growth was monitored by turbidity measurement (McFarland units). The data are representative of three independent experiments. (B) Differential sensitivity of M. tuberculosis wild-type and the ctpC null mutant to free zinc. M. tuberculosis wild-type (GC1237), a ctpC null mutant ( ctpC ::Kan R ), or the cosmid-complemented strain (I437) were allowed to grow in 7H9-ADC medium containing 0.5 mM ZnSO 4 or without zinc supplementation (Control). Bacterial growth was monitored by plating on agar and counting CFU. The data are representative of two independent experiments. (C) Differential ability of M. tuberculosis wild-type and the ctpC null mutant to replicate in human macrophages. M. tuberculosis wild-type (GC1237), a ctpC null mutant ( ctpC ::Kan R ), or a plasmid-complemented strain (CP) was used to infect human macrophages at a multiplicity of infection of one mycobacteria per ten cells. After 4 hr, cells were washed and incubated in fresh medium for 5 days. The data shown are means ±SD of intracellular CFU counts in an experiment carried out in triplicate and analyzed with Student's t test. ∗ p

    Journal: Cell Host & Microbe

    Article Title: Mycobacterial P1-Type ATPases Mediate Resistance to Zinc Poisoning in Human Macrophages

    doi: 10.1016/j.chom.2011.08.006

    Figure Lengend Snippet: CtpC Is Involved in Zinc Detoxification and Contributes to the Intracellular Survival of M. tuberculosis (A) Differential sensitivity of M. tuberculosis wild-type and the ctpC null mutant to free zinc. M. tuberculosis wild-type (GC1237), a ctpC null mutant ( ctpC ::Kan R ), or the cosmid-complemented strain (I437) was allowed to grow in 7H9-ADC medium containing 0.1 mM ZnSO 4 or without zinc supplementation (Control). Bacterial growth was monitored by turbidity measurement (McFarland units). The data are representative of three independent experiments. (B) Differential sensitivity of M. tuberculosis wild-type and the ctpC null mutant to free zinc. M. tuberculosis wild-type (GC1237), a ctpC null mutant ( ctpC ::Kan R ), or the cosmid-complemented strain (I437) were allowed to grow in 7H9-ADC medium containing 0.5 mM ZnSO 4 or without zinc supplementation (Control). Bacterial growth was monitored by plating on agar and counting CFU. The data are representative of two independent experiments. (C) Differential ability of M. tuberculosis wild-type and the ctpC null mutant to replicate in human macrophages. M. tuberculosis wild-type (GC1237), a ctpC null mutant ( ctpC ::Kan R ), or a plasmid-complemented strain (CP) was used to infect human macrophages at a multiplicity of infection of one mycobacteria per ten cells. After 4 hr, cells were washed and incubated in fresh medium for 5 days. The data shown are means ±SD of intracellular CFU counts in an experiment carried out in triplicate and analyzed with Student's t test. ∗ p

    Article Snippet: Mycobacteria were grown in Middlebrook 7H9 culture medium (Difco) supplemented with 10% albumin-dextrose-catalase (ADC, Difco), 0.05% Tween-80 (Sigma) in Sauton's medium supplemented with 0.05% Tween 80, or on Middlebrook 7H11 agar (Difco) supplemented with 10% oleic acid-ADC (OADC, Difco).

    Techniques: Mutagenesis, Plasmid Preparation, Infection, Incubation

    In vitro stability of pBP10 in M. smegmatis and Mtb in the absence of antibiotic selection. ( a ) Total number of M. smegmatis -pBP10, accounting for serial dilutions to show ongoing culture expansion. ( b ) Frequency of M. smegmatis -pBP10 plasmid-containing bacteria. ( c ) Frequency of plasmid carriage in M. smegmatis -pBP10 for log-phase cultures versus generations (calculated as (log(OD 600 t / OD 600 ( t − 1)) / log(2)). ( d ) Total number of M. tuberculosis -pBP10. ( e ) Frequency of M. tuberculosis -pBP10 plasmid-containing bacteria. ( f ) Frequency of plasmid carriage in M. tuberculosis -pBP10 for log-phase cultures versus generations. Data (means ± s.d.) are shown for log-phase cultures in 7H9, 1:1 7H9 to water, 1:3 7H9 to water, 7H9 without shaking and 7H9 in 2% O 2 . Representative data are shown for two to five experiments performed in triplicate, except for the 2% O 2 experiment, which was performed once.

    Journal: Nature medicine

    Article Title: A replication clock for Mycobacterium tuberculosis

    doi: 10.1038/nm.1915

    Figure Lengend Snippet: In vitro stability of pBP10 in M. smegmatis and Mtb in the absence of antibiotic selection. ( a ) Total number of M. smegmatis -pBP10, accounting for serial dilutions to show ongoing culture expansion. ( b ) Frequency of M. smegmatis -pBP10 plasmid-containing bacteria. ( c ) Frequency of plasmid carriage in M. smegmatis -pBP10 for log-phase cultures versus generations (calculated as (log(OD 600 t / OD 600 ( t − 1)) / log(2)). ( d ) Total number of M. tuberculosis -pBP10. ( e ) Frequency of M. tuberculosis -pBP10 plasmid-containing bacteria. ( f ) Frequency of plasmid carriage in M. tuberculosis -pBP10 for log-phase cultures versus generations. Data (means ± s.d.) are shown for log-phase cultures in 7H9, 1:1 7H9 to water, 1:3 7H9 to water, 7H9 without shaking and 7H9 in 2% O 2 . Representative data are shown for two to five experiments performed in triplicate, except for the 2% O 2 experiment, which was performed once.

    Article Snippet: For the hypoxic experiments, we maintained Mtb in 7H9 medium in 96-well plates placed inside airtight bags with a Gaspak EZ Anaerobe Container System Sachet (Becton Dickinson) and a Gaspak Dry Anaerobic Indicator Strip (Becton Dickinson).

    Techniques: In Vitro, Selection, Plasmid Preparation

    Survival of Erdman strain of M. tb in 7H9 media. M. tb grown in 7H9 media containing no additives (A) , M. tb grown in 7H9 media containing NAC (B) , M. tb grown in 7H9 media containing INH and INH+ NAC (C) , M. tb grown in 7H9 media containing RIF and RIF + NAC (D) , and M. tb grown in 7H9 media containing EMB and EMB+ NAC (E) . There was a significant increase in bacterial numbers at 15 days when M. tb was grown in 7H9 in the absence of any additives (A) . There was a significant reduction in the bacterial numbers at 15 days when M. tb was grown in 7H9 in the presence of NAC (B) , INH and INH+NAC (C) , RIF and RIF+NAC (D) and EMB and EMB+NAC (E) . Data represent means ±SE from experiments performed in triplicate. * p

    Journal: Frontiers in Immunology

    Article Title: The Synergistic Effects of the Glutathione Precursor, NAC and First-Line Antibiotics in the Granulomatous Response Against Mycobacterium tuberculosis

    doi: 10.3389/fimmu.2018.02069

    Figure Lengend Snippet: Survival of Erdman strain of M. tb in 7H9 media. M. tb grown in 7H9 media containing no additives (A) , M. tb grown in 7H9 media containing NAC (B) , M. tb grown in 7H9 media containing INH and INH+ NAC (C) , M. tb grown in 7H9 media containing RIF and RIF + NAC (D) , and M. tb grown in 7H9 media containing EMB and EMB+ NAC (E) . There was a significant increase in bacterial numbers at 15 days when M. tb was grown in 7H9 in the absence of any additives (A) . There was a significant reduction in the bacterial numbers at 15 days when M. tb was grown in 7H9 in the presence of NAC (B) , INH and INH+NAC (C) , RIF and RIF+NAC (D) and EMB and EMB+NAC (E) . Data represent means ±SE from experiments performed in triplicate. * p

    Article Snippet: M. tb was cultured in 7H9 media medium (Hi Media, Santa Maria, CA, USA) supplemented with albumin dextrose complex (ADC) (GEMINI, USA) and incubated at 37°C until the bacteria reached logarithmic phase of growth.

    Techniques:

    MAb 24c5 ELISA analysis and binding to glycogen. (A) Splenic fusion resulted in MAb 24c5, an IgG1 able to recognize M. tuberculosis GC. Each data point represents the average of three measurements. (B) MAb 24c5 binding to dilute concentrations of GC in the M. tuberculosis GC ELISA. Each data point represents the average of three measurements. (C) MAb 24c5 binding to M. tuberculosis Erdman day-20 7H9 medium (MTB), cell culture supernatant extract (INTPHSE), GC, AM, lipoarabinomannan (LAM), phosphatidylinositol mannoside (PIM), fast-growing mycobacterial lipomannan (LM), mycolyl-arabinogalactan-peptidoglycan complex (mAGP), and total lipid fraction (TLF). Each bar represents the average of two measurements. (D) Effects of proteinase K on binding of MAb 24c5 to M. tuberculosis Erdman day-20 7H9 medium. (E and F) MAb 24c5 assayed for reactivity to type VIII (slipper limpet) (viii) (E) and type IX (ix) (bovine liver) (F) glycogens. Each data point represents the average of three measurements. Error bars show the standard deviations of the means.

    Journal: Infection and Immunity

    Article Title: Glucan Is a Component of the Mycobacterium tuberculosis Surface That Is Expressed In Vitro and In Vivo

    doi: 10.1128/IAI.70.5.2566-2575.2002

    Figure Lengend Snippet: MAb 24c5 ELISA analysis and binding to glycogen. (A) Splenic fusion resulted in MAb 24c5, an IgG1 able to recognize M. tuberculosis GC. Each data point represents the average of three measurements. (B) MAb 24c5 binding to dilute concentrations of GC in the M. tuberculosis GC ELISA. Each data point represents the average of three measurements. (C) MAb 24c5 binding to M. tuberculosis Erdman day-20 7H9 medium (MTB), cell culture supernatant extract (INTPHSE), GC, AM, lipoarabinomannan (LAM), phosphatidylinositol mannoside (PIM), fast-growing mycobacterial lipomannan (LM), mycolyl-arabinogalactan-peptidoglycan complex (mAGP), and total lipid fraction (TLF). Each bar represents the average of two measurements. (D) Effects of proteinase K on binding of MAb 24c5 to M. tuberculosis Erdman day-20 7H9 medium. (E and F) MAb 24c5 assayed for reactivity to type VIII (slipper limpet) (viii) (E) and type IX (ix) (bovine liver) (F) glycogens. Each data point represents the average of three measurements. Error bars show the standard deviations of the means.

    Article Snippet: Mycobacterial cultures were grown with (7H9-T medium) or without (7H9 medium) 0.05% Tween 80 (Sigma) for 11, 20, and 25 days in 490-cm2 roller bottles (Corning Inc., Corning, N.Y.) in a 5% CO2 incubator at 37°C at pathogen level 3, as described by Schwebach et al. ( ).

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Cell Culture, Laser Capture Microdissection

    Survival of Δ cpsA under in vitro stress conditions. ( A ) Growth curve of H37Rv, Δ cpsA , and Δ cpsA::cpsA grown in 7H9 medium. ( B ) Survival of H37Rv, Δ cpsA , and Δ katG in the presence of 5 mM H 2 O 2 . Samples were plated after 30, 60, 90, 120, and 180 min. No colonies were observed for Δ katG after 30 min (#). *** P = 0.0005. ( C ) Survival of H37Rv and Δ cpsA after the addition of 200 μM diethylenetriamine/nitric oxide (DETA-NO) every 24 h for 3 d. ns, not significant; Student’s t test. ( D ) In vitro growth curves of H37Rv and Δ cpsA in 7H9 medium under different pHs (range 5.0–7.5). Data are representative of two experiments. ( E ) Sensitivity of H37Rv or Δ cpsA in the presence of lysozyme. The indicated concentration of lysozyme was added to 7H11 plates. Data are representative of two independent experiments. * P ≤ 0.05, *** P ≤ 0.0005; Student’s t test. ( F ) Sensitivity of H37Rv and Δ cpsA to SDS. The indicated bacterial numbers were spotted on 7H11 agar plates with SDS (0.005%, 0.01%) or without SDS. Pictures were taken 15 d after spotting. Data represent the results of three independent experiments.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Mycobacterium tuberculosis is protected from NADPH oxidase and LC3-associated phagocytosis by the LCP protein CpsA

    doi: 10.1073/pnas.1707792114

    Figure Lengend Snippet: Survival of Δ cpsA under in vitro stress conditions. ( A ) Growth curve of H37Rv, Δ cpsA , and Δ cpsA::cpsA grown in 7H9 medium. ( B ) Survival of H37Rv, Δ cpsA , and Δ katG in the presence of 5 mM H 2 O 2 . Samples were plated after 30, 60, 90, 120, and 180 min. No colonies were observed for Δ katG after 30 min (#). *** P = 0.0005. ( C ) Survival of H37Rv and Δ cpsA after the addition of 200 μM diethylenetriamine/nitric oxide (DETA-NO) every 24 h for 3 d. ns, not significant; Student’s t test. ( D ) In vitro growth curves of H37Rv and Δ cpsA in 7H9 medium under different pHs (range 5.0–7.5). Data are representative of two experiments. ( E ) Sensitivity of H37Rv or Δ cpsA in the presence of lysozyme. The indicated concentration of lysozyme was added to 7H11 plates. Data are representative of two independent experiments. * P ≤ 0.05, *** P ≤ 0.0005; Student’s t test. ( F ) Sensitivity of H37Rv and Δ cpsA to SDS. The indicated bacterial numbers were spotted on 7H11 agar plates with SDS (0.005%, 0.01%) or without SDS. Pictures were taken 15 d after spotting. Data represent the results of three independent experiments.

    Article Snippet: Mtb strains were grown at 37 °C in 7H9 medium (Middlebrook 7H9 broth; Difco) supplemented with 0.05% Tween 80 (Sigma), BBL Middlebrook OADC Enrichment, and 0.2% glycerol (Sigma).

    Techniques: In Vitro, Concentration Assay

    The Δ cpsA mutant has normal growth characteristics and antibiotic susceptibility. ( A ) H37Rv and Δ cpsA were grown on 7H11 plates for 14 and 19 d. ( B ) H37Rv and Δ cpsA growing in 7H9 medium are shown immediately after shaking and after being held static. ( C ) Equivalent optical densities of H37Rv and Δ cpsA were plated on 7H11 medium, and the number of bacteria was enumerated by cfu. ( D ) H37Rv and Δ cpsA were grown under biofilm-promoting conditions. ( E ) The fluorescence associated with ethidium bromide uptake was measured over time in H37Rv and Δ cpsA . ( F ) Growth of H37Rv and Δ cpsA in the presence of the indicated concentrations of antibiotics.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Mycobacterium tuberculosis is protected from NADPH oxidase and LC3-associated phagocytosis by the LCP protein CpsA

    doi: 10.1073/pnas.1707792114

    Figure Lengend Snippet: The Δ cpsA mutant has normal growth characteristics and antibiotic susceptibility. ( A ) H37Rv and Δ cpsA were grown on 7H11 plates for 14 and 19 d. ( B ) H37Rv and Δ cpsA growing in 7H9 medium are shown immediately after shaking and after being held static. ( C ) Equivalent optical densities of H37Rv and Δ cpsA were plated on 7H11 medium, and the number of bacteria was enumerated by cfu. ( D ) H37Rv and Δ cpsA were grown under biofilm-promoting conditions. ( E ) The fluorescence associated with ethidium bromide uptake was measured over time in H37Rv and Δ cpsA . ( F ) Growth of H37Rv and Δ cpsA in the presence of the indicated concentrations of antibiotics.

    Article Snippet: Mtb strains were grown at 37 °C in 7H9 medium (Middlebrook 7H9 broth; Difco) supplemented with 0.05% Tween 80 (Sigma), BBL Middlebrook OADC Enrichment, and 0.2% glycerol (Sigma).

    Techniques: Mutagenesis, Fluorescence

    Growth analysis of M. smegmatis mc 2 155 and knockout mutants (two single mutants [Δms33 and Δms32] and one double mutant [Δms33/32]). The strains were inoculated (McFarland number 0.5) into Middlebrook 7H9 medium (supplemented with OADC enrichment and appropriate antibiotics), and the CFU per milliliter were counted every 3 h for 42 h. The graph was plotted on semilogarithmic base 2 scales ( y axis). The values are means ± SDs from three independent experiments. The growth rate and doubling time were calculated for each replicate using the linear part of the log phase.

    Journal: Journal of Bacteriology

    Article Title: Two dd-Carboxypeptidases from Mycobacterium smegmatis Affect Cell Surface Properties through Regulation of Peptidoglycan Cross-Linking and Glycopeptidolipids

    doi: 10.1128/JB.00760-17

    Figure Lengend Snippet: Growth analysis of M. smegmatis mc 2 155 and knockout mutants (two single mutants [Δms33 and Δms32] and one double mutant [Δms33/32]). The strains were inoculated (McFarland number 0.5) into Middlebrook 7H9 medium (supplemented with OADC enrichment and appropriate antibiotics), and the CFU per milliliter were counted every 3 h for 42 h. The graph was plotted on semilogarithmic base 2 scales ( y axis). The values are means ± SDs from three independent experiments. The growth rate and doubling time were calculated for each replicate using the linear part of the log phase.

    Article Snippet: Mycobacterial strains ( ) were grown in Middlebrook (MB) 7H9 medium supplemented with 10% (vol/vol) oleic acid-albumin-dextrose-catalase (OADC; Difco, MD), 0.05% (vol/vol) Tween 80, and glycerol.

    Techniques: Knock-Out, Mutagenesis

    Loss of ICD2 results in glutamate auxotrophy and impaired viability. ( a ) Growth (OD 600 ) of wild-type, Δ icd and Δ icd attB ::P np icd -complemented strains of M. smegmatis in Middlebrook 7H9 medium. Solid lines, culture density (OD 600 ). Dashed lines, glutamate concentration in culture medium. ( b ) Growth (OD 600 ) of wild-type, Δ icd and Δ icd attB ::P np icd -complemented strains of M. smegmatis in minimal medium supplemented with glucose and devoid of glutamate. Data are means±s.d. ( n =3 independent experiments). ( c ) Intracellular metabolites in wild-type, Δ icd and Δ icd attB ::P np icd -complemented strains of M. smegmatis 24 h after transferring cells into minimal medium supplemented with glucose and devoid of glutamate. Data are means±s.d. ( n =3 independent experiments). CIT/ICT, citrate/isocitrate; α-KG, alpha-ketoglutarate; GLU, glutamate. ( d , e ) Growth (OD 600 ) of wild-type, Δ icd1 , Δ icd2 and Δ icd2 attB ::P np icd2 -complemented strains of M. bovis BCG in minimal medium supplemented with glucose and glutamate ( d ) or without glutamate ( e ). ( f ) Survival (CFU) of wild-type, Δ icd and Δ icd attB ::P np icd -complemented strains of M. smegmatis in minimal medium supplemented with glucose and devoid of glutamate. Data are means±s.d. ( n =3 independent experiments each performed in triplicate). ( g ) Survival (CFU) of wild-type, Δ icd2 and Δ icd2 attB ::P np icd2 -complemented strains of M. bovis BCG in minimal medium supplemented with glucose and devoid of glutamate. Data are means±s.d. ( n =3 independent experiments each performed in triplicate).

    Journal: Nature Communications

    Article Title: A rheostat mechanism governs the bifurcation of carbon flux in mycobacteria

    doi: 10.1038/ncomms12527

    Figure Lengend Snippet: Loss of ICD2 results in glutamate auxotrophy and impaired viability. ( a ) Growth (OD 600 ) of wild-type, Δ icd and Δ icd attB ::P np icd -complemented strains of M. smegmatis in Middlebrook 7H9 medium. Solid lines, culture density (OD 600 ). Dashed lines, glutamate concentration in culture medium. ( b ) Growth (OD 600 ) of wild-type, Δ icd and Δ icd attB ::P np icd -complemented strains of M. smegmatis in minimal medium supplemented with glucose and devoid of glutamate. Data are means±s.d. ( n =3 independent experiments). ( c ) Intracellular metabolites in wild-type, Δ icd and Δ icd attB ::P np icd -complemented strains of M. smegmatis 24 h after transferring cells into minimal medium supplemented with glucose and devoid of glutamate. Data are means±s.d. ( n =3 independent experiments). CIT/ICT, citrate/isocitrate; α-KG, alpha-ketoglutarate; GLU, glutamate. ( d , e ) Growth (OD 600 ) of wild-type, Δ icd1 , Δ icd2 and Δ icd2 attB ::P np icd2 -complemented strains of M. bovis BCG in minimal medium supplemented with glucose and glutamate ( d ) or without glutamate ( e ). ( f ) Survival (CFU) of wild-type, Δ icd and Δ icd attB ::P np icd -complemented strains of M. smegmatis in minimal medium supplemented with glucose and devoid of glutamate. Data are means±s.d. ( n =3 independent experiments each performed in triplicate). ( g ) Survival (CFU) of wild-type, Δ icd2 and Δ icd2 attB ::P np icd2 -complemented strains of M. bovis BCG in minimal medium supplemented with glucose and devoid of glutamate. Data are means±s.d. ( n =3 independent experiments each performed in triplicate).

    Article Snippet: Viability of M. smegmatis and M. bovis BCG cultures was determined by plating serially diluted aliquots of cultures on LB solid medium for M. smegmatis or Middlebrook 7H9 solid medium supplemented with 0.5% glycerol and 10% oleic acid–albumin–dextrose–catalase (0.5 g l−1 oleic acid, 50 g l−1 bovine serum albumin fraction V, 20 g l−1 glucose, 40 mg l−1 catalase, 8.5 g l−1 NaCl) for M. bovis BCG.

    Techniques: Concentration Assay, Transferring

    51 V NMR (78.9 MHz) spectra are shown of solution of decavanadate (10 mM V 10 , 100 mM V-atoms). The samples are from the bottom up diluted V 10 stock solution (100 mM V-atom) at pH 3.1; 10 mM V 10 in the presence of 0.48 mM and 0.97 mM citrate at pH 2.8 and 2.2, respectively; 10 mM V 10 in the presence of 24 mM P i at pH 6.9; and finally 10 mM V 10 in the presence of Middlebrook 7H9 broth medium supplemented with 10% ADC enrichment (5% BSA, 2% dextrose, 5% catalase), glycerol (0.2%, v/v) and Tween 80 (0.05%, v/v) recorded both in the absence and the presence of a capillary reference of 100 mM Na 3 VO 4 .

    Journal: Frontiers in Chemistry

    Article Title: Decavanadate Inhibits Mycobacterial Growth More Potently Than Other Oxovanadates

    doi: 10.3389/fchem.2018.00519

    Figure Lengend Snippet: 51 V NMR (78.9 MHz) spectra are shown of solution of decavanadate (10 mM V 10 , 100 mM V-atoms). The samples are from the bottom up diluted V 10 stock solution (100 mM V-atom) at pH 3.1; 10 mM V 10 in the presence of 0.48 mM and 0.97 mM citrate at pH 2.8 and 2.2, respectively; 10 mM V 10 in the presence of 24 mM P i at pH 6.9; and finally 10 mM V 10 in the presence of Middlebrook 7H9 broth medium supplemented with 10% ADC enrichment (5% BSA, 2% dextrose, 5% catalase), glycerol (0.2%, v/v) and Tween 80 (0.05%, v/v) recorded both in the absence and the presence of a capillary reference of 100 mM Na 3 VO 4 .

    Article Snippet: The bacteria were grown in 7H9 Middlebrook medium with the addition of D-pantothenate (24 mg/L) at 37°C to an optical density at 600 nm (OD600nm ) of 0.6–0.8.

    Techniques: Nuclear Magnetic Resonance

    51 V NMR (78.9 MHz) spectra are shown of solution of colorless oxovanadate (40 mM V 1 , 40 mM V-atoms). The samples are from the bottom up diluted V 1 stock solution (40 mM V-atom) at pH 8.3; 10 mM V 10 in the presence of 24 mM P i at pH 8.1; 10 mM V 10 in the presence of 0.48 mM citrate at pH 6.3, and finally 10 mM V 10 in the presence of Middlebrook 7H9 broth medium supplemented with 10% ADC enrichment (5% BSA, 2% dextrose, 5% catalase), glycerol (0.2%, v/v) and Tween 80 (0.05%, v/v) recorded both in the absence and the presence of a capillary reference of 100 mM Na 3 VO 4 . The spectrum labeled Reference is of the capillary reference alone (top spectrum). The key to the signals: V-oligomers, V 1 monomer; V 2 , dimer; V 4 , tetramer; V 5 , pentamer; VCit, V-citrate complex; PV, vanadate-phosphate complex.

    Journal: Frontiers in Chemistry

    Article Title: Decavanadate Inhibits Mycobacterial Growth More Potently Than Other Oxovanadates

    doi: 10.3389/fchem.2018.00519

    Figure Lengend Snippet: 51 V NMR (78.9 MHz) spectra are shown of solution of colorless oxovanadate (40 mM V 1 , 40 mM V-atoms). The samples are from the bottom up diluted V 1 stock solution (40 mM V-atom) at pH 8.3; 10 mM V 10 in the presence of 24 mM P i at pH 8.1; 10 mM V 10 in the presence of 0.48 mM citrate at pH 6.3, and finally 10 mM V 10 in the presence of Middlebrook 7H9 broth medium supplemented with 10% ADC enrichment (5% BSA, 2% dextrose, 5% catalase), glycerol (0.2%, v/v) and Tween 80 (0.05%, v/v) recorded both in the absence and the presence of a capillary reference of 100 mM Na 3 VO 4 . The spectrum labeled Reference is of the capillary reference alone (top spectrum). The key to the signals: V-oligomers, V 1 monomer; V 2 , dimer; V 4 , tetramer; V 5 , pentamer; VCit, V-citrate complex; PV, vanadate-phosphate complex.

    Article Snippet: The bacteria were grown in 7H9 Middlebrook medium with the addition of D-pantothenate (24 mg/L) at 37°C to an optical density at 600 nm (OD600nm ) of 0.6–0.8.

    Techniques: Nuclear Magnetic Resonance, Labeling

    Loss of mce operons increases clumping. Aggregation of M. smegmatis mc 2 155 and its Δ6 mce ] Panel A: MH; Panel B: Middlebrook 7H9-ADS-glycerol. Panel C: determination of the

    Journal: Microbes and infection / Institut Pasteur

    Article Title: Impact of the deletion of the six mce operons in Mycobacterium smegmatis

    doi: 10.1016/j.micinf.2012.01.007

    Figure Lengend Snippet: Loss of mce operons increases clumping. Aggregation of M. smegmatis mc 2 155 and its Δ6 mce ] Panel A: MH; Panel B: Middlebrook 7H9-ADS-glycerol. Panel C: determination of the

    Article Snippet: The first examination of the deletion mutants by naked eye failed to reveal any noticeable phenotype affecting size or morphology of the colonies on Middlebrook 7H9 agar medium (data not shown).

    Techniques:

    SufR TB protein is essential for Mtb to survive under stress conditions. ( A and B ) Survival of wild-type, mutant, and complemented strains in 7H9 enriched medium under oxidative (5 mM H 2 O 2 for 6 hours) and nitrosative stress (200 μM DETA-NO for 48 hours), respectively. ( C) Growth of the ∆ sufR TB strain relative to the wild-type strain in mouse bone marrow-derived macrophages. Macrophages were infected at an MOI of 1, and a relative growth difference was estimated by counting colony-forming units at day 0 and day 7 after plating. ( D ) Biofilm formation was observed in wild-type and mutant strains in Sauton’s medium over a period of 4 weeks. Significant differences observed in the groups are marked (unpaired two-tailed t test, *P

    Journal: Scientific Reports

    Article Title: Iron homeostasis in Mycobacterium tuberculosis is essential for persistence

    doi: 10.1038/s41598-018-35012-3

    Figure Lengend Snippet: SufR TB protein is essential for Mtb to survive under stress conditions. ( A and B ) Survival of wild-type, mutant, and complemented strains in 7H9 enriched medium under oxidative (5 mM H 2 O 2 for 6 hours) and nitrosative stress (200 μM DETA-NO for 48 hours), respectively. ( C) Growth of the ∆ sufR TB strain relative to the wild-type strain in mouse bone marrow-derived macrophages. Macrophages were infected at an MOI of 1, and a relative growth difference was estimated by counting colony-forming units at day 0 and day 7 after plating. ( D ) Biofilm formation was observed in wild-type and mutant strains in Sauton’s medium over a period of 4 weeks. Significant differences observed in the groups are marked (unpaired two-tailed t test, *P

    Article Snippet: Wild-type, ∆sufR TB , and ∆sufR TB :pJEB ∆sufR TB strains of Mtb were grown in Middlebrook 7H9 supplemented medium.

    Techniques: Mutagenesis, Derivative Assay, Infection, Two Tailed Test

    Generation of Mtb sufR deletion mutant. (A ) Schematic representation of the homologous recombination between the upstream and downstream region of Rv1460 gene cloned in the pJM1 suicidal vector and H37Rv genome. ( B ) sufR TB (Rv1460) gene replaced by the hygromycin cassette in the H37Rv genome due to homologous recombination, generating a deletion mutant. ( C ) For Southern blot analysis, genomic DNA was isolated from different strains by using the CTAB method. DNA (5 µg) was digested with NcoI and transferred onto nitrocellulose membranes and probed with a DIG-labelled specific probe, upstream to sufR TB gene. ( D ) Quantitative PCR depicting the upregulation of the ISC operon in the mutant strain grown in 7H9 medium enriched with 10% OADC and 0.05% Tween 80.

    Journal: Scientific Reports

    Article Title: Iron homeostasis in Mycobacterium tuberculosis is essential for persistence

    doi: 10.1038/s41598-018-35012-3

    Figure Lengend Snippet: Generation of Mtb sufR deletion mutant. (A ) Schematic representation of the homologous recombination between the upstream and downstream region of Rv1460 gene cloned in the pJM1 suicidal vector and H37Rv genome. ( B ) sufR TB (Rv1460) gene replaced by the hygromycin cassette in the H37Rv genome due to homologous recombination, generating a deletion mutant. ( C ) For Southern blot analysis, genomic DNA was isolated from different strains by using the CTAB method. DNA (5 µg) was digested with NcoI and transferred onto nitrocellulose membranes and probed with a DIG-labelled specific probe, upstream to sufR TB gene. ( D ) Quantitative PCR depicting the upregulation of the ISC operon in the mutant strain grown in 7H9 medium enriched with 10% OADC and 0.05% Tween 80.

    Article Snippet: Wild-type, ∆sufR TB , and ∆sufR TB :pJEB ∆sufR TB strains of Mtb were grown in Middlebrook 7H9 supplemented medium.

    Techniques: Mutagenesis, Homologous Recombination, Clone Assay, Plasmid Preparation, Southern Blot, Isolation, Real-time Polymerase Chain Reaction

    ∆ sufR TB fails to grow under a low iron condition. ( A ) Growth curve of wild-type, mutant, and complemented strains in 7H9 enriched medium. ( B and C ) Growth curve of wild-type (H37Rv) and ∆ sufR TB strains in minimal medium supplemented with 0.1% glycerol at varying concentrations of ferric ammonium citrate. ( D ) Growth analysis of wild-type, mutant, and complemented strains in 7H9 enriched medium containing different concentrations of the Fe chelator, depicting growth attenuation in the mutant strain treated with 0.32 mM of 2, 2′-bipyridyl.

    Journal: Scientific Reports

    Article Title: Iron homeostasis in Mycobacterium tuberculosis is essential for persistence

    doi: 10.1038/s41598-018-35012-3

    Figure Lengend Snippet: ∆ sufR TB fails to grow under a low iron condition. ( A ) Growth curve of wild-type, mutant, and complemented strains in 7H9 enriched medium. ( B and C ) Growth curve of wild-type (H37Rv) and ∆ sufR TB strains in minimal medium supplemented with 0.1% glycerol at varying concentrations of ferric ammonium citrate. ( D ) Growth analysis of wild-type, mutant, and complemented strains in 7H9 enriched medium containing different concentrations of the Fe chelator, depicting growth attenuation in the mutant strain treated with 0.32 mM of 2, 2′-bipyridyl.

    Article Snippet: Wild-type, ∆sufR TB , and ∆sufR TB :pJEB ∆sufR TB strains of Mtb were grown in Middlebrook 7H9 supplemented medium.

    Techniques: Mutagenesis

    Growth of M. smegmatis in various nitrogen sources. Wild-type SMR5 and glnR deletion strain MH1 were grown in the presence of the indicated substances (10 mM final concentration) as sole nitrogen source. Standard 7H9 medium was used as positive control; 7H9 lacking any nitrogen source as negative control (7H9-N). (A) Putative nitrogen sources deduced from microarray results, (B) putative nitrogen sources used by closely related species.

    Journal: BMC Research Notes

    Article Title: Nitrogen starvation-induced transcriptome alterations and influence of transcription regulator mutants in Mycobacterium smegmatis

    doi: 10.1186/1756-0500-6-482

    Figure Lengend Snippet: Growth of M. smegmatis in various nitrogen sources. Wild-type SMR5 and glnR deletion strain MH1 were grown in the presence of the indicated substances (10 mM final concentration) as sole nitrogen source. Standard 7H9 medium was used as positive control; 7H9 lacking any nitrogen source as negative control (7H9-N). (A) Putative nitrogen sources deduced from microarray results, (B) putative nitrogen sources used by closely related species.

    Article Snippet: Mycobacterial strains were grown in Middlebrook 7H9 liquid medium (Difco Laboratories; per 900 ml approx.

    Techniques: Concentration Assay, Positive Control, Negative Control, Microarray

    Mass spectrometry analysis of MAs. MAs were extracted from mid-log-phase liquid 7H9 medium cultures. Samples were analyzed via a QTRAP 4000 mass spectrometer. (A) Individual sums of C 26 α-, C 24 α-, C 26 keto-, and C 24 keto-MA profiles in

    Journal: Infection and Immunity

    Article Title: An ethA-ethR-Deficient Mycobacterium bovis BCG Mutant Displays Increased Adherence to Mammalian Cells and Greater Persistence In Vivo, Which Correlate with Altered Mycolic Acid Composition

    doi: 10.1128/IAI.01332-13

    Figure Lengend Snippet: Mass spectrometry analysis of MAs. MAs were extracted from mid-log-phase liquid 7H9 medium cultures. Samples were analyzed via a QTRAP 4000 mass spectrometer. (A) Individual sums of C 26 α-, C 24 α-, C 26 keto-, and C 24 keto-MA profiles in

    Article Snippet: Wild-type (WT) M. bovis BCG (Pasteur strain ATCC 35734) and derivative strains were grown at 37°C in Middlebrook liquid 7H9 medium (Difco) or on 7H11 agar supplemented with ADS (0.5% bovine serum albumin fraction V, 0.2% dextrose, 0.85% saline) enrichment, 0.05% Tween 80, and 0.2% glycerol with appropriate antibiotics (80 μg/ml hygromycin [Roche], 20 μg/ml kanamycin [Sigma]).

    Techniques: Mass Spectrometry

    Kinetics and inducibility of fructose, glucose, and glycerol uptake by M. smegmatis . (A) M. smegmatis mc 2 155 was grown in Middlebrook 7H9 medium in the presence of 2% glycerol (open circles) or 2% fructose (closed circles). Accumulation

    Journal:

    Article Title: A Genomic View of Sugar Transport in Mycobacterium smegmatis and Mycobacterium tuberculosis ▿

    doi: 10.1128/JB.00257-07

    Figure Lengend Snippet: Kinetics and inducibility of fructose, glucose, and glycerol uptake by M. smegmatis . (A) M. smegmatis mc 2 155 was grown in Middlebrook 7H9 medium in the presence of 2% glycerol (open circles) or 2% fructose (closed circles). Accumulation

    Article Snippet: M. smegmatis mc2 155 was grown in liquid cultures using Middlebrook 7H9 medium (Difco) supplemented with 0.2% glycerol and 0.05% Tween 80 or minimal Hartmans-de Bont (HB) medium ( ) at 37°C.

    Techniques:

    Cell surface of Msmeg-PE11 is highly hydrophobic. ( a ) Pellicle formation of Msmeg-pVV or Msmeg-PE11 was monitored by growing standing cultures of the strains without shaking in Middlebrook 7H9 medium in absence of Tween 80 at 37 °C for various time points. ( b ) Biofilm formation was quantified by crystal violet staining for which Msmeg-pVV or Msmeg-PE11 cells were washed, stained with 1% crystal violet and ethanol extract was spectrophotometrically measured at 570 nm. ( c ) Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 medium with (upper panel) or without (lower panel) 0.05% Tween-80 at 37 °C with shaking for 24 h and 48 h. Cultures were then allowed to settle at room temperature for 30 min. ( d ) In another experiment, Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 broth with congo red (100 μg/ml) and 0.05% Tween 80 for 24 h, 48 h and 72 h at 37 °C. Cells were next washed and re-suspended in acetone. Congo red in the supernatant was spectrophotometrically measured at 490 nm. Data are representative of mean ± SD of three different experiments.

    Journal: Scientific Reports

    Article Title: PE11, a PE/PPE family protein of Mycobacterium tuberculosis is involved in cell wall remodeling and virulence

    doi: 10.1038/srep21624

    Figure Lengend Snippet: Cell surface of Msmeg-PE11 is highly hydrophobic. ( a ) Pellicle formation of Msmeg-pVV or Msmeg-PE11 was monitored by growing standing cultures of the strains without shaking in Middlebrook 7H9 medium in absence of Tween 80 at 37 °C for various time points. ( b ) Biofilm formation was quantified by crystal violet staining for which Msmeg-pVV or Msmeg-PE11 cells were washed, stained with 1% crystal violet and ethanol extract was spectrophotometrically measured at 570 nm. ( c ) Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 medium with (upper panel) or without (lower panel) 0.05% Tween-80 at 37 °C with shaking for 24 h and 48 h. Cultures were then allowed to settle at room temperature for 30 min. ( d ) In another experiment, Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 broth with congo red (100 μg/ml) and 0.05% Tween 80 for 24 h, 48 h and 72 h at 37 °C. Cells were next washed and re-suspended in acetone. Congo red in the supernatant was spectrophotometrically measured at 490 nm. Data are representative of mean ± SD of three different experiments.

    Article Snippet: M. smegmatis culture and transformation M. smegmatis mc2 155 bacteria were grown in Middlebrook 7H9 medium (BD Difco, USA) supplemented with 10% OADC (HiMedia, India), 0.5% glycerol, and 0.05% Tween 80 (Fisher Scientific, USA).

    Techniques: Staining, Cell Culture

    Genetic organization, growth analysis, bacterial survival, and Mtb Δ esxL mutant construction. A , schematic representation of e sxL in the M. tuberculosis H37Rv genome. RAW 264.7 ( B ) and THP-1 ( C ) were infected with M. smegmatis ( Msm ) pSMT3 and recombinant M. smegmatis esxL strains. The cells were lysed, and intracellular survival was determined 1, 8, and 24 h post-infection by a cfu assay. D , in vitro growth curve of the M. smegmatis WT, M. smegmatis pSMT3, and recombinant M. smegmatis esxL was determined by growing bacteria in 7H9 medium and measuring OD ( O.D 600 nm ) . E , extracellular expression of the esxL transcript was measured by qRT-PCR after growing M. smegmatis esxL in vitro for 4, 12, and 24 h. RNA was isolated at the respective time points. cDNA was synthesized, and the expression of esxL was determined using qRT-PCR. Transcript levels are represented relative to mRNA -fold change of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. F , intracellular expression of esxL transcript was measured by qRT-PCR. RNA was isolated from M. smegmatis esxL -infected macrophages at different time points. cDNA was synthesized, and the expression of esxL was determined using qRT PCR. Transcript levels are represented relative to mRNA level of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. G , schematic representation of construction of Mtb Δ esxL mutant by homologous recombination. The location of primers used for the confirmation of deletion mutant generation is depicted. H , confirmation of Mtb Δ esxL mutant generation. F1 and R2 primers were designed beyond the flanks, whereas R1 and F2 primers anneal to sacB-hyg r cassette. PCR using F1 and R1 is expected to give no product with the M. tuberculosis ( lane 1 ) and ∼1.3 kb with the Mtb Δ esxL ( lane 2 ); F2-R2 primer sets were expected to give no product with M. tuberculosis and ∼1.5 kb in Mtb Δ esxL mutant. Amplification of udgB with gene-specific primers was performed as a control. The experiments were performed in triplicate ( n = 3). Results are shown as mean ± S.D. ( error bars ). *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ns , not significant.

    Journal: The Journal of Biological Chemistry

    Article Title: Mycobacterium tuberculosis EsxL inhibits MHC-II expression by promoting hypermethylation in class-II transactivator loci in macrophages

    doi: 10.1074/jbc.M117.775205

    Figure Lengend Snippet: Genetic organization, growth analysis, bacterial survival, and Mtb Δ esxL mutant construction. A , schematic representation of e sxL in the M. tuberculosis H37Rv genome. RAW 264.7 ( B ) and THP-1 ( C ) were infected with M. smegmatis ( Msm ) pSMT3 and recombinant M. smegmatis esxL strains. The cells were lysed, and intracellular survival was determined 1, 8, and 24 h post-infection by a cfu assay. D , in vitro growth curve of the M. smegmatis WT, M. smegmatis pSMT3, and recombinant M. smegmatis esxL was determined by growing bacteria in 7H9 medium and measuring OD ( O.D 600 nm ) . E , extracellular expression of the esxL transcript was measured by qRT-PCR after growing M. smegmatis esxL in vitro for 4, 12, and 24 h. RNA was isolated at the respective time points. cDNA was synthesized, and the expression of esxL was determined using qRT-PCR. Transcript levels are represented relative to mRNA -fold change of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. F , intracellular expression of esxL transcript was measured by qRT-PCR. RNA was isolated from M. smegmatis esxL -infected macrophages at different time points. cDNA was synthesized, and the expression of esxL was determined using qRT PCR. Transcript levels are represented relative to mRNA level of M. smegmatis esxL at 4 h, which is assigned a value of 1. The expression values were normalized with sigA. G , schematic representation of construction of Mtb Δ esxL mutant by homologous recombination. The location of primers used for the confirmation of deletion mutant generation is depicted. H , confirmation of Mtb Δ esxL mutant generation. F1 and R2 primers were designed beyond the flanks, whereas R1 and F2 primers anneal to sacB-hyg r cassette. PCR using F1 and R1 is expected to give no product with the M. tuberculosis ( lane 1 ) and ∼1.3 kb with the Mtb Δ esxL ( lane 2 ); F2-R2 primer sets were expected to give no product with M. tuberculosis and ∼1.5 kb in Mtb Δ esxL mutant. Amplification of udgB with gene-specific primers was performed as a control. The experiments were performed in triplicate ( n = 3). Results are shown as mean ± S.D. ( error bars ). *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ns , not significant.

    Article Snippet: Mycobacterium smegmatis mc2 155 was grown in Middlebrook's 7H9 broth medium (Difco) containing 0.05% Tween 80, 0.5% glucose, and 0.5% albumin at 37 °C on a shaker at 120 rpm.

    Techniques: Mutagenesis, Infection, Recombinant, Colony-forming Unit Assay, In Vitro, Expressing, Quantitative RT-PCR, Isolation, Synthesized, Homologous Recombination, Polymerase Chain Reaction, Amplification

    Growth curves of the WT, ΔMycP 3 mutant and the two complementation strains, ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms under Fe-free 7H9 ( a ), Fe-free Sauton’s media ( b ). The growth curves were done in triplicate, error bars show standard deviation

    Journal: BMC Research Notes

    Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro

    doi: 10.1186/s13104-017-2752-0

    Figure Lengend Snippet: Growth curves of the WT, ΔMycP 3 mutant and the two complementation strains, ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms under Fe-free 7H9 ( a ), Fe-free Sauton’s media ( b ). The growth curves were done in triplicate, error bars show standard deviation

    Article Snippet: M. smegmatis was cultured using both Middlebrook 7H9 liquid medium (Becton–Dickinson, USA, Catalogue No. 221832) and Difco 7H11 solid medium (Becton–Dickinson, USA, Catalogue No. 212304) both supplemented with 0.05% (v/v) Tween 80 (Sigma Aldrich, USA, Catalogue No. P1754), 0.5% (w/v) glucose (Sigma Aldrich, USA, Catalogue No. 47829), and 0.5% (v/v) glycerol (Sigma-Aldrich, USA, Catalogue No. G5516).

    Techniques: Mutagenesis, Mass Spectrometry, Standard Deviation

    The comparison of the intracellular iron levels in WT ms , ΔMycP3 ms , ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms strains under 7H9, Fe-depleted 7H9 and Fe rescued 7H9 media. The error bars show standard error of the mean (n = 4). The p values obtained using two-way ANOVA statistical analysis between different culturing conditions for all four strains are smaller than 0.0001 ( *** ), an example is shown for the WT ms

    Journal: BMC Research Notes

    Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro

    doi: 10.1186/s13104-017-2752-0

    Figure Lengend Snippet: The comparison of the intracellular iron levels in WT ms , ΔMycP3 ms , ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms strains under 7H9, Fe-depleted 7H9 and Fe rescued 7H9 media. The error bars show standard error of the mean (n = 4). The p values obtained using two-way ANOVA statistical analysis between different culturing conditions for all four strains are smaller than 0.0001 ( *** ), an example is shown for the WT ms

    Article Snippet: M. smegmatis was cultured using both Middlebrook 7H9 liquid medium (Becton–Dickinson, USA, Catalogue No. 221832) and Difco 7H11 solid medium (Becton–Dickinson, USA, Catalogue No. 212304) both supplemented with 0.05% (v/v) Tween 80 (Sigma Aldrich, USA, Catalogue No. P1754), 0.5% (w/v) glucose (Sigma Aldrich, USA, Catalogue No. 47829), and 0.5% (v/v) glycerol (Sigma-Aldrich, USA, Catalogue No. G5516).

    Techniques: Mass Spectrometry

    Growth curves of the WT, ΔMycP 3 mutant and the two complementation strains, ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms under Fe-depleted 7H9 ( a ), and Fe-depleted Sauton’s media ( b ). The growth curves were done in triplicate, error bars show standard deviation

    Journal: BMC Research Notes

    Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro

    doi: 10.1186/s13104-017-2752-0

    Figure Lengend Snippet: Growth curves of the WT, ΔMycP 3 mutant and the two complementation strains, ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms under Fe-depleted 7H9 ( a ), and Fe-depleted Sauton’s media ( b ). The growth curves were done in triplicate, error bars show standard deviation

    Article Snippet: M. smegmatis was cultured using both Middlebrook 7H9 liquid medium (Becton–Dickinson, USA, Catalogue No. 221832) and Difco 7H11 solid medium (Becton–Dickinson, USA, Catalogue No. 212304) both supplemented with 0.05% (v/v) Tween 80 (Sigma Aldrich, USA, Catalogue No. P1754), 0.5% (w/v) glucose (Sigma Aldrich, USA, Catalogue No. 47829), and 0.5% (v/v) glycerol (Sigma-Aldrich, USA, Catalogue No. G5516).

    Techniques: Mutagenesis, Mass Spectrometry, Standard Deviation

    Gene expression analysis of mycP 3 in WT ms , ΔMycP 3ms , ΔMycP 3ms ::Pr1MycP 3 , and ΔMycP 3ms ::Pr2MycP 3 strains under normal 7H9 (iron rich), Fe-free 7H9, and Fe-rescued 7H9 media. The results were normalized against the RNA copy number of sigA . The p values obtained using two-way ANOVA statistical analysis (n = 3) (*p

    Journal: BMC Research Notes

    Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro

    doi: 10.1186/s13104-017-2752-0

    Figure Lengend Snippet: Gene expression analysis of mycP 3 in WT ms , ΔMycP 3ms , ΔMycP 3ms ::Pr1MycP 3 , and ΔMycP 3ms ::Pr2MycP 3 strains under normal 7H9 (iron rich), Fe-free 7H9, and Fe-rescued 7H9 media. The results were normalized against the RNA copy number of sigA . The p values obtained using two-way ANOVA statistical analysis (n = 3) (*p

    Article Snippet: M. smegmatis was cultured using both Middlebrook 7H9 liquid medium (Becton–Dickinson, USA, Catalogue No. 221832) and Difco 7H11 solid medium (Becton–Dickinson, USA, Catalogue No. 212304) both supplemented with 0.05% (v/v) Tween 80 (Sigma Aldrich, USA, Catalogue No. P1754), 0.5% (w/v) glucose (Sigma Aldrich, USA, Catalogue No. 47829), and 0.5% (v/v) glycerol (Sigma-Aldrich, USA, Catalogue No. G5516).

    Techniques: Expressing, Mass Spectrometry

    Anti- Mycobacterium bovis capacity of rHBD3 protein to A549 cells. a M. bovis growth curve after incubation with different concentrations of HBD-3. Negative control is M. bovis with Middle Brook 7H9 broth. Positive control is M. bovis with streptomycin (1000 U/ml). b Mycobacteria invasion tests. Yellow spots in the figure are Mycobacteria . c Cell apoptosis analysis of A549 cells infected by M. bovis with different treatments. d The data of cell apoptosis and death ratio analysis of A549 cells infected by M. bovis with different treatments. All the experiments were replicated three times and the changes are presented as mean ± SEM. P values

    Journal: AMB Express

    Article Title: Expression of recombinant HBD3 protein that reduces Mycobacterial infection capacity

    doi: 10.1186/s13568-018-0573-8

    Figure Lengend Snippet: Anti- Mycobacterium bovis capacity of rHBD3 protein to A549 cells. a M. bovis growth curve after incubation with different concentrations of HBD-3. Negative control is M. bovis with Middle Brook 7H9 broth. Positive control is M. bovis with streptomycin (1000 U/ml). b Mycobacteria invasion tests. Yellow spots in the figure are Mycobacteria . c Cell apoptosis analysis of A549 cells infected by M. bovis with different treatments. d The data of cell apoptosis and death ratio analysis of A549 cells infected by M. bovis with different treatments. All the experiments were replicated three times and the changes are presented as mean ± SEM. P values

    Article Snippet: The colonies were then transferred to Middlebrook 7H9 modified medium (Difco Laboratories, Detroit, MI, USA) for 20 days.

    Techniques: Incubation, Negative Control, Positive Control, Infection

    Growth of RGM within A. polyphaga trophozoites. M. gilvum (A), M. rhodesiae (B), and M. thermoresistibile (C) were cocultured with the free-living amoeba A. polyphaga (black bars), cultivated in PAS medium (gray bars), and cultivated in 7H9 complete medium

    Journal: Applied and Environmental Microbiology

    Article Title: Mycobacterium gilvum Illustrates Size-Correlated Relationships between Mycobacteria and Acanthamoeba polyphaga

    doi: 10.1128/AEM.03765-12

    Figure Lengend Snippet: Growth of RGM within A. polyphaga trophozoites. M. gilvum (A), M. rhodesiae (B), and M. thermoresistibile (C) were cocultured with the free-living amoeba A. polyphaga (black bars), cultivated in PAS medium (gray bars), and cultivated in 7H9 complete medium

    Article Snippet: Mycobacteria were cultured in Middlebrook 7H9 liquid medium (Sigma-Aldrich, Lyon, France) and subcultured at 37°C on Middlebrook and Cohn 7H10 agar (Becton, Dickinson, Le Pont de Claix, France) for 3 days.

    Techniques:

    Mycobacterial size in 7H9 medium. The sizes of M. gilvum (A), M. senegalense (B), M. conceptionense (C), M. rhodesiae (D), M. thermoresistibile (E), M. chelonae (F), M. smegmatis (G), M. abscessus (H), and M. fortuitum subsp. fortuitum (I) were determined

    Journal: Applied and Environmental Microbiology

    Article Title: Mycobacterium gilvum Illustrates Size-Correlated Relationships between Mycobacteria and Acanthamoeba polyphaga

    doi: 10.1128/AEM.03765-12

    Figure Lengend Snippet: Mycobacterial size in 7H9 medium. The sizes of M. gilvum (A), M. senegalense (B), M. conceptionense (C), M. rhodesiae (D), M. thermoresistibile (E), M. chelonae (F), M. smegmatis (G), M. abscessus (H), and M. fortuitum subsp. fortuitum (I) were determined

    Article Snippet: Mycobacteria were cultured in Middlebrook 7H9 liquid medium (Sigma-Aldrich, Lyon, France) and subcultured at 37°C on Middlebrook and Cohn 7H10 agar (Becton, Dickinson, Le Pont de Claix, France) for 3 days.

    Techniques:

    Growth of M. tuberculosis strains in liquid and solid media. (A) Wild-type M. tuberculosis H37Rv, Δ cuvA Mt , and complemented Δ cuvA (Δ cuvA Mt /C) strains were grown in 7H9-ADC-Tw liquid medium with 0.2% glucose, and the OD 600 was measured daily. (B) Growth of the same strains on agar plates containing MM plus 0.2% glucose or MM plus 0.01% cholesterol. Serial 10-fold dilutions were spotted and incubated at 37°C, and photographs were taken after 1 month of incubation. In both panels, data shown are from one experiment that was repeated at least twice with similar results. Glc, 0.2% glucose; Chol., 0.01% cholesterol.

    Journal: Infection and Immunity

    Article Title: Mycobacterial Gene cuvA Is Required for Optimal Nutrient Utilization and Virulence

    doi: 10.1128/IAI.02207-14

    Figure Lengend Snippet: Growth of M. tuberculosis strains in liquid and solid media. (A) Wild-type M. tuberculosis H37Rv, Δ cuvA Mt , and complemented Δ cuvA (Δ cuvA Mt /C) strains were grown in 7H9-ADC-Tw liquid medium with 0.2% glucose, and the OD 600 was measured daily. (B) Growth of the same strains on agar plates containing MM plus 0.2% glucose or MM plus 0.01% cholesterol. Serial 10-fold dilutions were spotted and incubated at 37°C, and photographs were taken after 1 month of incubation. In both panels, data shown are from one experiment that was repeated at least twice with similar results. Glc, 0.2% glucose; Chol., 0.01% cholesterol.

    Article Snippet: For measurements of growth on solid medium, cells grown in Middlebrook 7H9 liquid medium to an optical density at 600 nm (OD600 ) of 0.5 were washed in phosphate-buffered saline containing 0.05% tyloxapol (PBS-Tx), and 10-fold serial dilutions in PBS were spotted on agar plates containing MM as described above (without tyloxapol) plus a 0.01% concentration of the carbon source.

    Techniques: Incubation, Gas Chromatography

    FAs of the K10 (Type C) and 2349/06-1 (Type S) isolates of Map that showed statistically significant differences in abundance between the two isolates . BOMAC and MOCL-4 cells were infected with the K10 and 2349/06-1 isolates of Map at MOI of 1:10. At 4 h p.i., the medium containing the extracellular bacteria was collected, centrifuged at 2000 g for 15 min and the FAMEs of the bacterial pellets extracted and analyzed by GC. FAs of the K10 and 2349/06-1 isolates of Map grown in Middlebrook 7H9 medium that showed statistically significant differences in abundance are also included in the figure. Relative amount of each FA for each experimental condition (7H9 medium or extracellular) is presented as the percentage of the total FAs content. Statistically significant differences are indicated with an asterisk.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mycobacterium avium subsp. paratuberculosis (Map) Fatty Acids Profile Is Strain-Dependent and Changes Upon Host Macrophages Infection

    doi: 10.3389/fcimb.2017.00089

    Figure Lengend Snippet: FAs of the K10 (Type C) and 2349/06-1 (Type S) isolates of Map that showed statistically significant differences in abundance between the two isolates . BOMAC and MOCL-4 cells were infected with the K10 and 2349/06-1 isolates of Map at MOI of 1:10. At 4 h p.i., the medium containing the extracellular bacteria was collected, centrifuged at 2000 g for 15 min and the FAMEs of the bacterial pellets extracted and analyzed by GC. FAs of the K10 and 2349/06-1 isolates of Map grown in Middlebrook 7H9 medium that showed statistically significant differences in abundance are also included in the figure. Relative amount of each FA for each experimental condition (7H9 medium or extracellular) is presented as the percentage of the total FAs content. Statistically significant differences are indicated with an asterisk.

    Article Snippet: The relative FAs composition of the two isolates recovered from infected BOMAC and MOCL-4 cells was determined by gas chromatography and compared with that of extracellular bacteria and that of bacteria grown in Middlebrook 7H9 medium.

    Techniques: Infection

    (A) Resistance of M. smegmatis to CuOOH is dependent in part upon Ohr. Cells were cultured to late stationary phase (17 days) in Middlebrook 7H9 medium. Cells were then suspended in the same medium at an A 600 of 0.1 and incubated with CuOOH for 2 h at

    Journal: Journal of Bacteriology

    Article Title: Organic Hydroperoxide Resistance Protein and Ergothioneine Compensate for Loss of Mycothiol in Mycobacterium smegmatis Mutants ▿ Mutants ▿ †

    doi: 10.1128/JB.01402-10

    Figure Lengend Snippet: (A) Resistance of M. smegmatis to CuOOH is dependent in part upon Ohr. Cells were cultured to late stationary phase (17 days) in Middlebrook 7H9 medium. Cells were then suspended in the same medium at an A 600 of 0.1 and incubated with CuOOH for 2 h at

    Article Snippet: M. smegmatis mc2 155 was also grown on Middlebrook 7H9 solid medium (1.8% Difco agar), with 0.5% glycerol supplemented with OADC or 1% glucose.

    Techniques: Cell Culture, Incubation