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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.
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1) Product Images from "Nitrogen starvation-induced transcriptome alterations and influence of transcription regulator mutants in Mycobacterium smegmatis"

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

Journal: BMC Research Notes

doi: 10.1186/1756-0500-6-482

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.
Figure Legend 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.

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

2) Product Images from "In Vitro Model of Mycobacterial Growth Arrest Using Nitric Oxide with Limited Air ▿"

Article Title: In Vitro Model of Mycobacterial Growth Arrest Using Nitric Oxide with Limited Air ▿

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00442-08

Effect of DETA-NO on growth of cultured M. bovis BCG. Bacteria growing exponentially in 7H9 medium were transferred to evacuated (Vacutainer) tubes at mid-log phase, and at time zero either the cells were left untreated (open circles) or DETA-NO was added
Figure Legend Snippet: Effect of DETA-NO on growth of cultured M. bovis BCG. Bacteria growing exponentially in 7H9 medium were transferred to evacuated (Vacutainer) tubes at mid-log phase, and at time zero either the cells were left untreated (open circles) or DETA-NO was added

Techniques Used: Cell Culture

3) Product Images from "Chlorine Disinfection of Atypical Mycobacteria Isolated from a Water Distribution System"

Article Title: Chlorine Disinfection of Atypical Mycobacteria Isolated from a Water Distribution System

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.68.3.1025-1032.2002

Effects of pH on the rate of inactivation of M. gordonae . Experimental conditions were a temperature of 25°C and an initial chlorine concentration of 0.5 mg/liter. Cells were grown in Middlebrook 7H9-Tween medium. No, initial number of CFU; N, number of CFU at each time point. Experiments were conducted with different phosphate (0.05 M) buffers within the pH range of 6.0 to 8.0. Slopes were calculated as follows: pH 6, y = 0.11 x ; pH 7, y = 0.09 x ; and pH 8, y = 0.02 x . R 2 , correlation coefficient values.
Figure Legend Snippet: Effects of pH on the rate of inactivation of M. gordonae . Experimental conditions were a temperature of 25°C and an initial chlorine concentration of 0.5 mg/liter. Cells were grown in Middlebrook 7H9-Tween medium. No, initial number of CFU; N, number of CFU at each time point. Experiments were conducted with different phosphate (0.05 M) buffers within the pH range of 6.0 to 8.0. Slopes were calculated as follows: pH 6, y = 0.11 x ; pH 7, y = 0.09 x ; and pH 8, y = 0.02 x . R 2 , correlation coefficient values.

Techniques Used: Concentration Assay

Effects of temperature on chlorine inactivation of M. gordonae . Experimental conditions were pH 7 and an initial chlorine concentration of 0.5 mg/liter. Cells were grown in Middlebrook 7H9-Tween medium. The chlorine susceptibility of M. gordonae was analyzed at 4, 16, and 25°C. No, initial number of CFU; N, number of CFU at each time point. Symbols: ▪, 4°C ( y = 0.01 x ); ▴, 16°C ( y = 0.02 x ); ⧫, 25°C ( y = 0.09 x ). R 2 , correlation coefficient values.
Figure Legend Snippet: Effects of temperature on chlorine inactivation of M. gordonae . Experimental conditions were pH 7 and an initial chlorine concentration of 0.5 mg/liter. Cells were grown in Middlebrook 7H9-Tween medium. The chlorine susceptibility of M. gordonae was analyzed at 4, 16, and 25°C. No, initial number of CFU; N, number of CFU at each time point. Symbols: ▪, 4°C ( y = 0.01 x ); ▴, 16°C ( y = 0.02 x ); ⧫, 25°C ( y = 0.09 x ). R 2 , correlation coefficient values.

Techniques Used: Concentration Assay

4) Product Images from "Octahedral ruthenium (II) polypyridyl complexes as antimicrobial agents against mycobacterium"

Article Title: Octahedral ruthenium (II) polypyridyl complexes as antimicrobial agents against mycobacterium

Journal: PeerJ

doi: 10.7717/peerj.3252

Complex 2 (A) was bacteriostatic and complex 3 (B) was bactericidal against M. smegmatis . Bacterial cells were inoculated 7H9 medium, and cultured without any drug or in the presence of complex 2 or 3 at various concentrations. At the indicated time points, aliquots of cell suspension were transferred and plated on drug-free 7H9 medium after 24 more hours of incubation.
Figure Legend Snippet: Complex 2 (A) was bacteriostatic and complex 3 (B) was bactericidal against M. smegmatis . Bacterial cells were inoculated 7H9 medium, and cultured without any drug or in the presence of complex 2 or 3 at various concentrations. At the indicated time points, aliquots of cell suspension were transferred and plated on drug-free 7H9 medium after 24 more hours of incubation.

Techniques Used: Cell Culture, Incubation

5) Product Images from "Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro"

Article Title: Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro

Journal: Infection and Immunity

doi:

Effect of exogenous H 2 O 2 on mycobacterial viability in cell-free Middlebrook 7H9 medium. H 2 O 2 (at the concentrations shown) was added to growing bacteria, and the number of CFU was determined as described in Materials and Methods. Results are expressed as percent survival relative to baseline ± SEM and are from one representative experiment with six replicate cultures. The numbers of CFU used for the experiment (and expressed as 100% in the figure) were 4.7 × 10 5 for H37Rv, 3.2 × 10 5 for H37Rv Inh r , and 2.8 × 10 5 for H37Rv(pMH59).
Figure Legend Snippet: Effect of exogenous H 2 O 2 on mycobacterial viability in cell-free Middlebrook 7H9 medium. H 2 O 2 (at the concentrations shown) was added to growing bacteria, and the number of CFU was determined as described in Materials and Methods. Results are expressed as percent survival relative to baseline ± SEM and are from one representative experiment with six replicate cultures. The numbers of CFU used for the experiment (and expressed as 100% in the figure) were 4.7 × 10 5 for H37Rv, 3.2 × 10 5 for H37Rv Inh r , and 2.8 × 10 5 for H37Rv(pMH59).

Techniques Used:

6) Product Images from "Mycobacterial Gene cuvA Is Required for Optimal Nutrient Utilization and Virulence"

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

Journal: Infection and Immunity

doi: 10.1128/IAI.02207-14

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.
Figure Legend 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.

Techniques Used: Incubation, Gas Chromatography

7) Product Images from "Mycobacterium tuberculosis Requires Regulation of ESX-5 Secretion for Virulence in Irgm1-Deficient Mice"

Article Title: Mycobacterium tuberculosis Requires Regulation of ESX-5 Secretion for Virulence in Irgm1-Deficient Mice

Journal: Infection and Immunity

doi: 10.1128/IAI.00660-18

Deletion of the esx-5 RegX3 binding site prevents activation of ESX-5 secretion in response to P i limitation. (A and B) Transcript abundances of pe19 , espG 5 , and eccD 5 relative to sigA were determined by quantitative RT-PCR for the indicated strains grown to mid-logarithmic phase in P i -free 7H9 medium (A) or P i -replete 7H9 complete medium (B). Results are the means ± standard deviations for three biological replicates, each run in technical duplicate. *, P
Figure Legend Snippet: Deletion of the esx-5 RegX3 binding site prevents activation of ESX-5 secretion in response to P i limitation. (A and B) Transcript abundances of pe19 , espG 5 , and eccD 5 relative to sigA were determined by quantitative RT-PCR for the indicated strains grown to mid-logarithmic phase in P i -free 7H9 medium (A) or P i -replete 7H9 complete medium (B). Results are the means ± standard deviations for three biological replicates, each run in technical duplicate. *, P

Techniques Used: Binding Assay, Activation Assay, Quantitative RT-PCR

Mutation of the esx-5 RegX3 binding site suppresses overexpression of esx-5 genes and hypersecretion of EsxN by the Δ pstA1 mutant. (A and B) Transcript abundances of pe19 , espG 5 , eccD 5 , udgA , and mgtA relative to sigA were determined by quantitative RT-PCR for the indicated strains grown to mid-logarithmic phase in 7H9 complete medium. Results are the means ± standard deviations for three biological replicates, each run in technical duplicate. **, P
Figure Legend Snippet: Mutation of the esx-5 RegX3 binding site suppresses overexpression of esx-5 genes and hypersecretion of EsxN by the Δ pstA1 mutant. (A and B) Transcript abundances of pe19 , espG 5 , eccD 5 , udgA , and mgtA relative to sigA were determined by quantitative RT-PCR for the indicated strains grown to mid-logarithmic phase in 7H9 complete medium. Results are the means ± standard deviations for three biological replicates, each run in technical duplicate. **, P

Techniques Used: Mutagenesis, Binding Assay, Over Expression, Quantitative RT-PCR

8) Product Images from "The Mycobacterium tuberculosis relBE toxin:antitoxin genes are stress-responsive modules that regulate growth through translation inhibition"

Article Title: The Mycobacterium tuberculosis relBE toxin:antitoxin genes are stress-responsive modules that regulate growth through translation inhibition

Journal: Journal of microbiology (Seoul, Korea)

doi: 10.1007/s12275-015-5333-8

Over expression of relE Mtb inhibits growth and macromolecular synthesis. M. smegmatis was transformed with either pYA1611 (LIX32), pYA- 1611:: relE (LIX33) or pYA1611:: relBE (LIX34). Overnight cultures were diluted to an OD 600 of 0.01 in 7H9-Tw-Hyg. At an OD 600 of 0.1, cultures were split, washed, resuspended in 7H9-Tw-Hyg and to one culture 100 ng/ml ATc was added to induce gene expression. (A) Growth of LIX33 with (solid line) and without (dashed line) ATc inducer. At the indicated time points uninduced LIX33 were diluted and plated on LB-agar, whereas induced LIX33 were plated on LB-ATc media. (B-D) Pulse-chase experiments were conducted to monitor the incorporation of radiolabeled precursors into (B) protein, [35S-Met], (C) RNA, [3H-Urd] or (D) DNA, [3H-Thy] with time. The incorporation rates are expressed as the percent relative to the incorporation of precursor into the uninduced culture at each time point. The values presented are the averages of three independent experiments; error bars represent the standard error of the mean. For statistical analysis, two-way analysis of variance with Bonferroni post tests was used to obtain P values for each time point *, P
Figure Legend Snippet: Over expression of relE Mtb inhibits growth and macromolecular synthesis. M. smegmatis was transformed with either pYA1611 (LIX32), pYA- 1611:: relE (LIX33) or pYA1611:: relBE (LIX34). Overnight cultures were diluted to an OD 600 of 0.01 in 7H9-Tw-Hyg. At an OD 600 of 0.1, cultures were split, washed, resuspended in 7H9-Tw-Hyg and to one culture 100 ng/ml ATc was added to induce gene expression. (A) Growth of LIX33 with (solid line) and without (dashed line) ATc inducer. At the indicated time points uninduced LIX33 were diluted and plated on LB-agar, whereas induced LIX33 were plated on LB-ATc media. (B-D) Pulse-chase experiments were conducted to monitor the incorporation of radiolabeled precursors into (B) protein, [35S-Met], (C) RNA, [3H-Urd] or (D) DNA, [3H-Thy] with time. The incorporation rates are expressed as the percent relative to the incorporation of precursor into the uninduced culture at each time point. The values presented are the averages of three independent experiments; error bars represent the standard error of the mean. For statistical analysis, two-way analysis of variance with Bonferroni post tests was used to obtain P values for each time point *, P

Techniques Used: Over Expression, Transformation Assay, Expressing, Pulse Chase

9) Product Images from "A rheostat mechanism governs the bifurcation of carbon flux in mycobacteria"

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

Journal: Nature Communications

doi: 10.1038/ncomms12527

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).
Figure Legend 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).

Techniques Used: Concentration Assay, Transferring

10) Product Images from "The Mycobacterium tuberculosis MmpL11 Cell Wall Lipid Transporter Is Important for Biofilm Formation, Intracellular Growth, and Nonreplicating Persistence"

Article Title: The Mycobacterium tuberculosis MmpL11 Cell Wall Lipid Transporter Is Important for Biofilm Formation, Intracellular Growth, and Nonreplicating Persistence

Journal: Infection and Immunity

doi: 10.1128/IAI.00131-17

M. tuberculosis mmpL11 mutant bacteria are shorter than the wild type in biofilms. Shown are representative images (left) and quantification of the lengths (right) of M. tuberculosis strains grown in either Sauton's medium lacking Tween 80, to promote biofilm formation (A), Sauton's medium containing Tween 80 (B), or 7H9 medium containing Tween 80 (C). The differences between the wild type and the mutant were significant by Student's t test (***, P
Figure Legend Snippet: M. tuberculosis mmpL11 mutant bacteria are shorter than the wild type in biofilms. Shown are representative images (left) and quantification of the lengths (right) of M. tuberculosis strains grown in either Sauton's medium lacking Tween 80, to promote biofilm formation (A), Sauton's medium containing Tween 80 (B), or 7H9 medium containing Tween 80 (C). The differences between the wild type and the mutant were significant by Student's t test (***, P

Techniques Used: Mutagenesis

Transmission electron microscopy reveals differences in ultrastructure between the wild-type and mmpL11 mutant M. tuberculosis strains grown in 7H9 medium containing Tween 80 (left) (planktonic) or Sauton's medium without Tween 80 (right) (biofilm). Arrows indicate crystal inclusions in mmpL11 mutants grown planktonically. Asterisks highlight the extracellular material that is enriched in the mmpL11 mutant relative to wild-type M. tuberculosis .
Figure Legend Snippet: Transmission electron microscopy reveals differences in ultrastructure between the wild-type and mmpL11 mutant M. tuberculosis strains grown in 7H9 medium containing Tween 80 (left) (planktonic) or Sauton's medium without Tween 80 (right) (biofilm). Arrows indicate crystal inclusions in mmpL11 mutants grown planktonically. Asterisks highlight the extracellular material that is enriched in the mmpL11 mutant relative to wild-type M. tuberculosis .

Techniques Used: Transmission Assay, Electron Microscopy, Mutagenesis

The M. tuberculosis mmpL11 mutant exhibits impaired biofilm formation that does not result from reduced replication in Sauton's medium. (A) Growth of M. tuberculosis wild-type and mmpL11 mutant strains in 7H9 medium. Average values and standard deviations for four biological replicates are shown. (B) Biofilm formation by M. tuberculosis wild-type, mmpL11 mutant, and complemented strains. Biofilms were cultured in 6-well plates for a total of 4 weeks. (C) Quantification of crystal violet staining of M. tuberculosis biofilms. Average values and standard deviations for three independent assays are shown. The difference between the wild type and the mutant was significant ( P ,
Figure Legend Snippet: The M. tuberculosis mmpL11 mutant exhibits impaired biofilm formation that does not result from reduced replication in Sauton's medium. (A) Growth of M. tuberculosis wild-type and mmpL11 mutant strains in 7H9 medium. Average values and standard deviations for four biological replicates are shown. (B) Biofilm formation by M. tuberculosis wild-type, mmpL11 mutant, and complemented strains. Biofilms were cultured in 6-well plates for a total of 4 weeks. (C) Quantification of crystal violet staining of M. tuberculosis biofilms. Average values and standard deviations for three independent assays are shown. The difference between the wild type and the mutant was significant ( P ,

Techniques Used: Mutagenesis, Cell Culture, Staining

11) Product Images from "Phosphate Starvation: a Novel Signal that Triggers ESX-5 Secretion in Mycobacterium tuberculosis"

Article Title: Phosphate Starvation: a Novel Signal that Triggers ESX-5 Secretion in Mycobacterium tuberculosis

Journal: Molecular microbiology

doi: 10.1111/mmi.13332

Induction of ESX-5 genes by phosphate limitation requires RegX3. Wild-type M. tuberculosis Erdman (WT), Δ regX3 , and Δ regX3 pND regX3 were cultured in P i -free 7H9 medium for 96 hours. RNA was extracted at 0, 24, 48, 72 and 96 hours. Abundance
Figure Legend Snippet: Induction of ESX-5 genes by phosphate limitation requires RegX3. Wild-type M. tuberculosis Erdman (WT), Δ regX3 , and Δ regX3 pND regX3 were cultured in P i -free 7H9 medium for 96 hours. RNA was extracted at 0, 24, 48, 72 and 96 hours. Abundance

Techniques Used: Cell Culture

12) Product Images from "MmpL11 Protein Transports Mycolic Acid-containing Lipids to the Mycobacterial Cell Wall and Contributes to Biofilm Formation in Mycobacterium smegmatis *"

Article Title: MmpL11 Protein Transports Mycolic Acid-containing Lipids to the Mycobacterial Cell Wall and Contributes to Biofilm Formation in Mycobacterium smegmatis *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M113.473371

The M. smegmatis mmpL11 mutant grows like the wild type in planktonic cultures but demonstrates impaired biofilm formation. A , growth of wild-type M. smegmatis and the mmpL11 mutant GP02 in complete 7H9 medium containing albumin/dextrose/saline supplements and 0.05% Tween 80 was followed over time. B , pellicle formation by the wild type (mc 2 155/pVV16), the mmpL11 mutant (GP02/pVV16), and the complemented mutant (GP02/pVV16 mmpL11 SM ). Bacteria were subcultured in glass tubes and allowed to grow at 37 °C for 5 days without shaking. At day 3, a thin film could be observed across the surface of the medium in the wild-type and complemented strains ( left ). This developed into a textured biofilm over the next 2 days. Attachment of bacteria to the glass tube and growth away from the surface could be observed from the side ( right ). C , bacteria in pellicle cultures depicted in A were enumerated. Biofilms were disrupted mechanically via syringing, and serial dilutions were plated. The mean ± S.D. of three independent experiments is shown. There was no significant difference in cfu/ml between strains. D , biofilm formation by the wild type (mc 2 155/pVV16), the mmpL11 mutant (GP02/pVV16), the complemented mutant (GP02/pVV16 mmpL11 SM ), and the heterologously complemented strain (GP02/pVV16 mmpL11 TB ). Equivalent numbers of bacteria were subcultured in liquid biofilm medium (Sauton's medium lacking Tween 80). Plates were incubated for 4 days at 30 °C. E , scanning electron microscopy of biofilms formed by wild-type, mmpL11 mutant, and complemented strains.
Figure Legend Snippet: The M. smegmatis mmpL11 mutant grows like the wild type in planktonic cultures but demonstrates impaired biofilm formation. A , growth of wild-type M. smegmatis and the mmpL11 mutant GP02 in complete 7H9 medium containing albumin/dextrose/saline supplements and 0.05% Tween 80 was followed over time. B , pellicle formation by the wild type (mc 2 155/pVV16), the mmpL11 mutant (GP02/pVV16), and the complemented mutant (GP02/pVV16 mmpL11 SM ). Bacteria were subcultured in glass tubes and allowed to grow at 37 °C for 5 days without shaking. At day 3, a thin film could be observed across the surface of the medium in the wild-type and complemented strains ( left ). This developed into a textured biofilm over the next 2 days. Attachment of bacteria to the glass tube and growth away from the surface could be observed from the side ( right ). C , bacteria in pellicle cultures depicted in A were enumerated. Biofilms were disrupted mechanically via syringing, and serial dilutions were plated. The mean ± S.D. of three independent experiments is shown. There was no significant difference in cfu/ml between strains. D , biofilm formation by the wild type (mc 2 155/pVV16), the mmpL11 mutant (GP02/pVV16), the complemented mutant (GP02/pVV16 mmpL11 SM ), and the heterologously complemented strain (GP02/pVV16 mmpL11 TB ). Equivalent numbers of bacteria were subcultured in liquid biofilm medium (Sauton's medium lacking Tween 80). Plates were incubated for 4 days at 30 °C. E , scanning electron microscopy of biofilms formed by wild-type, mmpL11 mutant, and complemented strains.

Techniques Used: Mutagenesis, Incubation, Electron Microscopy

13) Product Images from "Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic"

Article Title: Rapid Cytolysis of Mycobacterium tuberculosis by Faropenem, an Orally Bioavailable β-Lactam Antibiotic

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.03461-14

Real-time single-cell analysis of faropenem-mediated growth inhibition. An M. tuberculosis strain expressing GFP was cultured in a microfluidic device under a constant flow of 7H9 medium and imaged at 1-h intervals. (A and D) Faropenem (28 μg/ml; ∼21× MIC) was added to the flow medium at 96 to 264 h. (B and E) Meropenem plus clavulanate (MPC; 8 μg/ml and 2.5 μg/ml, respectively; ∼26× MIC) was added to the flow medium at 96 to 330 h. (C and F) Isoniazid (0.5 μg/ml; ∼20× MIC) was added to the flow medium at 96 to 340 h. (A to C) Images were recorded on fluorescence (green) and phase channels and merged. Numbers (lower right) indicate the elapsed times (in hours). Labels (upper right) indicate the presence or absence of antibiotic in the flow medium. (D to F) Single-cell growth rates were determined by measuring the projected areas of single cells ( n = 50) during the indicated time intervals and fitting exponential curves to the data.
Figure Legend Snippet: Real-time single-cell analysis of faropenem-mediated growth inhibition. An M. tuberculosis strain expressing GFP was cultured in a microfluidic device under a constant flow of 7H9 medium and imaged at 1-h intervals. (A and D) Faropenem (28 μg/ml; ∼21× MIC) was added to the flow medium at 96 to 264 h. (B and E) Meropenem plus clavulanate (MPC; 8 μg/ml and 2.5 μg/ml, respectively; ∼26× MIC) was added to the flow medium at 96 to 330 h. (C and F) Isoniazid (0.5 μg/ml; ∼20× MIC) was added to the flow medium at 96 to 340 h. (A to C) Images were recorded on fluorescence (green) and phase channels and merged. Numbers (lower right) indicate the elapsed times (in hours). Labels (upper right) indicate the presence or absence of antibiotic in the flow medium. (D to F) Single-cell growth rates were determined by measuring the projected areas of single cells ( n = 50) during the indicated time intervals and fitting exponential curves to the data.

Techniques Used: Single-cell Analysis, Inhibition, Expressing, Cell Culture, Flow Cytometry, Fluorescence

Activity of faropenem against extracellular and intracellular M. tuberculosis . (A) Bacteria expressing luciferase were incubated for 7 days in 7H9 medium containing different concentrations of faropenem (FPM) or faropenem plus clavulanate (FPMC). Growth was measured using a luciferase-based assay, and the data were normalized to the values obtained in the absence of antibiotic. Results are representative of those from four experiments. (B) Log-phase cultures were exposed for 6 days to different concentrations of faropenem and plated to measure the number of CFU. Values were normalized to those for the untreated controls. Results are representative of those from two experiments. (C) Log-phase cultures were exposed to antibiotics with (filled symbols) or without (empty symbols) 2.5 μg/ml clavulanate. Circles, meropenem (8 μg/ml; ∼26× MIC with clavulanate, ∼3× MIC without clavulanate); squares, faropenem (8 μg/ml; ∼6× MIC with or without clavulanate); triangles, faropenem (28 μg/ml; ∼21× MIC); crosses, 0.5 μg/ml isoniazid (∼20× MIC). At 0, 1, 2, 4, and 8 days after antibiotic addition, aliquots were washed and plated to measure the number of CFU. Results are means ± SEMs from three experiments. The rebound of the numbers of CFU at 8 days in the culture exposed to isoniazid was due to the outgrowth of drug-resistant variants. (D) The stability of compounds in 7H9 medium was measured by HPLC-UV analysis. Clav, clavulanic acid; MPM, meropenem. (E and F) Bacteria expressing GFP were grown in RAW macrophages and left untreated (closed circles) or treated with 56 μg/ml faropenem (open squares), 50 μg/ml meropenem plus 2.5 μg/ml clavulanate (open circles), or 0.75 μg/ml isoniazid (crosses). (E) Macrophage lysates were plated at 0 h and at 1, 3, and 6 days postinfection to measure the number of CFU. Data are plotted as percent survival normalized to the number of organisms at 0 h. (F) The fraction of macrophages (Mϕ) containing GFP-positive M. tuberculosis was measured by flow cytometry at 1, 3, and 6 days postinfection. INH, isoniazid; FPM, faropenem; MPMC, meropenem plus clavulanate. Results are means ± SEMs from three experiments.
Figure Legend Snippet: Activity of faropenem against extracellular and intracellular M. tuberculosis . (A) Bacteria expressing luciferase were incubated for 7 days in 7H9 medium containing different concentrations of faropenem (FPM) or faropenem plus clavulanate (FPMC). Growth was measured using a luciferase-based assay, and the data were normalized to the values obtained in the absence of antibiotic. Results are representative of those from four experiments. (B) Log-phase cultures were exposed for 6 days to different concentrations of faropenem and plated to measure the number of CFU. Values were normalized to those for the untreated controls. Results are representative of those from two experiments. (C) Log-phase cultures were exposed to antibiotics with (filled symbols) or without (empty symbols) 2.5 μg/ml clavulanate. Circles, meropenem (8 μg/ml; ∼26× MIC with clavulanate, ∼3× MIC without clavulanate); squares, faropenem (8 μg/ml; ∼6× MIC with or without clavulanate); triangles, faropenem (28 μg/ml; ∼21× MIC); crosses, 0.5 μg/ml isoniazid (∼20× MIC). At 0, 1, 2, 4, and 8 days after antibiotic addition, aliquots were washed and plated to measure the number of CFU. Results are means ± SEMs from three experiments. The rebound of the numbers of CFU at 8 days in the culture exposed to isoniazid was due to the outgrowth of drug-resistant variants. (D) The stability of compounds in 7H9 medium was measured by HPLC-UV analysis. Clav, clavulanic acid; MPM, meropenem. (E and F) Bacteria expressing GFP were grown in RAW macrophages and left untreated (closed circles) or treated with 56 μg/ml faropenem (open squares), 50 μg/ml meropenem plus 2.5 μg/ml clavulanate (open circles), or 0.75 μg/ml isoniazid (crosses). (E) Macrophage lysates were plated at 0 h and at 1, 3, and 6 days postinfection to measure the number of CFU. Data are plotted as percent survival normalized to the number of organisms at 0 h. (F) The fraction of macrophages (Mϕ) containing GFP-positive M. tuberculosis was measured by flow cytometry at 1, 3, and 6 days postinfection. INH, isoniazid; FPM, faropenem; MPMC, meropenem plus clavulanate. Results are means ± SEMs from three experiments.

Techniques Used: Activity Assay, Expressing, Luciferase, Incubation, High Performance Liquid Chromatography, Flow Cytometry, Cytometry

Detection of membrane potential in single cells of isoniazid-treated M. tuberculosis . Bacteria were cultured in a microfluidic device under a constant flow of 7H9 medium. Medium conditions were as follows: t = 0 to 96 h, no antibiotic; t = 96 to 340 h, addition of isoniazid (0.5 μg/ml; 20× MIC); t = 340 to 400 h, no antibiotic. As an endpoint assay, 3 μM DiOC 2 was added to the flow medium, and imaging was continued at 1-h intervals on the FITC and Texas Red fluorescence channels. Representative snapshots and the merged images from four random positions in the microfluidic device are shown in the four rows. In this assay, all cells exhibit green fluorescence, but higher cytosolic concentrations of the dye lead to self-association and a shift toward red fluorescence emission in cells with an intact membrane potential.
Figure Legend Snippet: Detection of membrane potential in single cells of isoniazid-treated M. tuberculosis . Bacteria were cultured in a microfluidic device under a constant flow of 7H9 medium. Medium conditions were as follows: t = 0 to 96 h, no antibiotic; t = 96 to 340 h, addition of isoniazid (0.5 μg/ml; 20× MIC); t = 340 to 400 h, no antibiotic. As an endpoint assay, 3 μM DiOC 2 was added to the flow medium, and imaging was continued at 1-h intervals on the FITC and Texas Red fluorescence channels. Representative snapshots and the merged images from four random positions in the microfluidic device are shown in the four rows. In this assay, all cells exhibit green fluorescence, but higher cytosolic concentrations of the dye lead to self-association and a shift toward red fluorescence emission in cells with an intact membrane potential.

Techniques Used: Cell Culture, Flow Cytometry, End Point Assay, Imaging, Fluorescence

14) Product Images from "Experimental selection of long-term intracellular mycobacteria"

Article Title: Experimental selection of long-term intracellular mycobacteria

Journal: Cellular Microbiology

doi: 10.1111/cmi.12303

Phenotypic and genetic changes of selected mycobacteria.A. In vitro growth of both ancestral and selected bacteria in complete 7H9 medium. Curves represent the mean ± S.E.M. of four independent experiments, (**) P ≤ 0.01 from two-tailed Student's t -test.B. Isotopic labelling of fatty acids according to the carbon length associated to neutral lipids after 1 day of incorporation of 13 C-acetate in complete 7H9 medium. The plot shows the incorporation of 13 C-acetate into neutral lipids in ANC and SEL bacteria ( x -axis).C. In vitro growth of ancestral and selected bacteria in minimal medium (Sauton) containing Tween 80, glycerol or glucose as a sole carbon source during 7 and 14 days. Data represent the mean ± S.E.M. of at least three independent experiments, (**) P ≤ 0.01 from two-tailed Student's t -test.D. Percentage of the fatty acid methyl esters (FAME) area associated to neutral lipids and corresponding to the indicated carbon length for ancestral and selected strains growing in minimal Sauton medium after 1 day of incorporation of 13 C-glucose as sole carbon source.E. Enrichment into fatty acids associated to neutral lipids after 9 days of growth in ancestral and selected strains growing in Sauton medium with 13 C-acetate as sole carbon source.F. Enrichment into fatty acids associated to neutral lipids after 9 days of growth in ancestral and selected strains growing in Sauton medium with 13 C-glucose as sole carbon source.
Figure Legend Snippet: Phenotypic and genetic changes of selected mycobacteria.A. In vitro growth of both ancestral and selected bacteria in complete 7H9 medium. Curves represent the mean ± S.E.M. of four independent experiments, (**) P ≤ 0.01 from two-tailed Student's t -test.B. Isotopic labelling of fatty acids according to the carbon length associated to neutral lipids after 1 day of incorporation of 13 C-acetate in complete 7H9 medium. The plot shows the incorporation of 13 C-acetate into neutral lipids in ANC and SEL bacteria ( x -axis).C. In vitro growth of ancestral and selected bacteria in minimal medium (Sauton) containing Tween 80, glycerol or glucose as a sole carbon source during 7 and 14 days. Data represent the mean ± S.E.M. of at least three independent experiments, (**) P ≤ 0.01 from two-tailed Student's t -test.D. Percentage of the fatty acid methyl esters (FAME) area associated to neutral lipids and corresponding to the indicated carbon length for ancestral and selected strains growing in minimal Sauton medium after 1 day of incorporation of 13 C-glucose as sole carbon source.E. Enrichment into fatty acids associated to neutral lipids after 9 days of growth in ancestral and selected strains growing in Sauton medium with 13 C-acetate as sole carbon source.F. Enrichment into fatty acids associated to neutral lipids after 9 days of growth in ancestral and selected strains growing in Sauton medium with 13 C-glucose as sole carbon source.

Techniques Used: In Vitro, Two Tailed Test, Isotopic Labeling

15) Product Images from "Mycobacterium Lysine ε-aminotransferase is a novel alarmone metabolism related persister gene via dysregulating the intracellular amino acid level"

Article Title: Mycobacterium Lysine ε-aminotransferase is a novel alarmone metabolism related persister gene via dysregulating the intracellular amino acid level

Journal: Scientific Reports

doi: 10.1038/srep19695

Construction of MSMEG_1764 knockout mutant and complement strains. ( A ) PCR verification of the construction MSMEG_1764 knockout strain. Lanes: 1.Wild-type MS; 2. MSMEG_1764 knockout strain. ( B ) Verify the transcription of MSMEG_1764 knockout strain by RT-PCR. Wild type and Δ lat Msm strains were grown at 37C in MB 7H9 liquid medium to an OD600 of 0.8–1.0. Total bacterial RNA was isolated and subjected to RT-PCR to detect the expression of the lat Msm gene. ( C ) Construction of Δ lat-Rv3290c strain; Lanes: 1. Knockout strain complement with pALACE- Rv3290 c; 2. Knockout strain complement with pALACE plasmid. ( D ) Western-blotting to confirm the expression of Rv3290c.Lysates were prepared from bacterial cells cultured as in ( B ), after 16 h induction and subjected to Western blotting to detect His-tagged Rv3290c protein using mouse anti-His antibody.
Figure Legend Snippet: Construction of MSMEG_1764 knockout mutant and complement strains. ( A ) PCR verification of the construction MSMEG_1764 knockout strain. Lanes: 1.Wild-type MS; 2. MSMEG_1764 knockout strain. ( B ) Verify the transcription of MSMEG_1764 knockout strain by RT-PCR. Wild type and Δ lat Msm strains were grown at 37C in MB 7H9 liquid medium to an OD600 of 0.8–1.0. Total bacterial RNA was isolated and subjected to RT-PCR to detect the expression of the lat Msm gene. ( C ) Construction of Δ lat-Rv3290c strain; Lanes: 1. Knockout strain complement with pALACE- Rv3290 c; 2. Knockout strain complement with pALACE plasmid. ( D ) Western-blotting to confirm the expression of Rv3290c.Lysates were prepared from bacterial cells cultured as in ( B ), after 16 h induction and subjected to Western blotting to detect His-tagged Rv3290c protein using mouse anti-His antibody.

Techniques Used: Knock-Out, Mutagenesis, Polymerase Chain Reaction, Mass Spectrometry, Reverse Transcription Polymerase Chain Reaction, Isolation, Expressing, Plasmid Preparation, Western Blot, Cell Culture

The expression of M. smegmatis lat under nutrient starvation. Cells of M. smegmatis were initially grown in 7H9 medium to log-phase and then washed by 1 × PBS and resuspended in 1 × PBS, incubate at 37 °C, 110 rpm. RNAs were extracted from bacteria harvested at indicated time points. RT-PCR was performed as described in Materials and Methods. Data are means ± s.d. of triplicates in one of at least three experiments.
Figure Legend Snippet: The expression of M. smegmatis lat under nutrient starvation. Cells of M. smegmatis were initially grown in 7H9 medium to log-phase and then washed by 1 × PBS and resuspended in 1 × PBS, incubate at 37 °C, 110 rpm. RNAs were extracted from bacteria harvested at indicated time points. RT-PCR was performed as described in Materials and Methods. Data are means ± s.d. of triplicates in one of at least three experiments.

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

16) Product Images from "Simultaneous Analysis of Multiple Mycobacterium tuberculosis Knockdown Mutants In Vitro and In Vivo"

Article Title: Simultaneous Analysis of Multiple Mycobacterium tuberculosis Knockdown Mutants In Vitro and In Vivo

Journal: PLoS ONE

doi: 10.1371/journal.pone.0015667

Growth of H37Rv, Erdman, rv3671c -TetON, prcBA -TetON, and icl -TetON in multi-strain liquid cultures. Multi-strain cultures were prepared, grown in 7H9 medium ( A ), Sauton's medium containing butyrate ( B ) or glycerol ( C ) and analyzed as indicated in Figure 2 . Black and clear symbols identify data generated from atc-containing or atc-free cultures, respectively. Data are averages of triplicate cultures and representative of at least two independent experiments. Error bars (representing standard deviations) cannot be seen because they are smaller than the data point symbols.
Figure Legend Snippet: Growth of H37Rv, Erdman, rv3671c -TetON, prcBA -TetON, and icl -TetON in multi-strain liquid cultures. Multi-strain cultures were prepared, grown in 7H9 medium ( A ), Sauton's medium containing butyrate ( B ) or glycerol ( C ) and analyzed as indicated in Figure 2 . Black and clear symbols identify data generated from atc-containing or atc-free cultures, respectively. Data are averages of triplicate cultures and representative of at least two independent experiments. Error bars (representing standard deviations) cannot be seen because they are smaller than the data point symbols.

Techniques Used: Generated

Impact of silencing rv3671c or prcBA on growth of Mtb in different liquid media. Growth of Mtb H37Rv (squares), rv3671c -TetON (triangles), prcBA -TetON (diamonds) without atc was analyzed in 7H9 medium ( A,D ), Sauton's medium containing butyrate ( B,E ) or glycerol ( C,F ) as the primary carbon source using optical density measurements at the indicated time points. Data represent results of three ( rv3671c -TetON) and two ( prcBA -TetON) independent experiments.
Figure Legend Snippet: Impact of silencing rv3671c or prcBA on growth of Mtb in different liquid media. Growth of Mtb H37Rv (squares), rv3671c -TetON (triangles), prcBA -TetON (diamonds) without atc was analyzed in 7H9 medium ( A,D ), Sauton's medium containing butyrate ( B,E ) or glycerol ( C,F ) as the primary carbon source using optical density measurements at the indicated time points. Data represent results of three ( rv3671c -TetON) and two ( prcBA -TetON) independent experiments.

Techniques Used:

17) Product Images from "MmpL11 Protein Transports Mycolic Acid-containing Lipids to the Mycobacterial Cell Wall and Contributes to Biofilm Formation in Mycobacterium smegmatis *"

Article Title: MmpL11 Protein Transports Mycolic Acid-containing Lipids to the Mycobacterial Cell Wall and Contributes to Biofilm Formation in Mycobacterium smegmatis *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M113.473371

The M. smegmatis mmpL11 mutant grows like the wild type in planktonic cultures but demonstrates impaired biofilm formation. A , growth of wild-type M. smegmatis and the mmpL11 mutant GP02 in complete 7H9 medium containing albumin/dextrose/saline supplements and 0.05% Tween 80 was followed over time. B , pellicle formation by the wild type (mc 2 155/pVV16), the mmpL11 mutant (GP02/pVV16), and the complemented mutant (GP02/pVV16 mmpL11 SM ). Bacteria were subcultured in glass tubes and allowed to grow at 37 °C for 5 days without shaking. At day 3, a thin film could be observed across the surface of the medium in the wild-type and complemented strains ( left ). This developed into a textured biofilm over the next 2 days. Attachment of bacteria to the glass tube and growth away from the surface could be observed from the side ( right ). C , bacteria in pellicle cultures depicted in A were enumerated. Biofilms were disrupted mechanically via syringing, and serial dilutions were plated. The mean ± S.D. of three independent experiments is shown. There was no significant difference in cfu/ml between strains. D , biofilm formation by the wild type (mc 2 155/pVV16), the mmpL11 mutant (GP02/pVV16), the complemented mutant (GP02/pVV16 mmpL11 SM ), and the heterologously complemented strain (GP02/pVV16 mmpL11 TB ). Equivalent numbers of bacteria were subcultured in liquid biofilm medium (Sauton's medium lacking Tween 80). Plates were incubated for 4 days at 30 °C. E , scanning electron microscopy of biofilms formed by wild-type, mmpL11 mutant, and complemented strains.
Figure Legend Snippet: The M. smegmatis mmpL11 mutant grows like the wild type in planktonic cultures but demonstrates impaired biofilm formation. A , growth of wild-type M. smegmatis and the mmpL11 mutant GP02 in complete 7H9 medium containing albumin/dextrose/saline supplements and 0.05% Tween 80 was followed over time. B , pellicle formation by the wild type (mc 2 155/pVV16), the mmpL11 mutant (GP02/pVV16), and the complemented mutant (GP02/pVV16 mmpL11 SM ). Bacteria were subcultured in glass tubes and allowed to grow at 37 °C for 5 days without shaking. At day 3, a thin film could be observed across the surface of the medium in the wild-type and complemented strains ( left ). This developed into a textured biofilm over the next 2 days. Attachment of bacteria to the glass tube and growth away from the surface could be observed from the side ( right ). C , bacteria in pellicle cultures depicted in A were enumerated. Biofilms were disrupted mechanically via syringing, and serial dilutions were plated. The mean ± S.D. of three independent experiments is shown. There was no significant difference in cfu/ml between strains. D , biofilm formation by the wild type (mc 2 155/pVV16), the mmpL11 mutant (GP02/pVV16), the complemented mutant (GP02/pVV16 mmpL11 SM ), and the heterologously complemented strain (GP02/pVV16 mmpL11 TB ). Equivalent numbers of bacteria were subcultured in liquid biofilm medium (Sauton's medium lacking Tween 80). Plates were incubated for 4 days at 30 °C. E , scanning electron microscopy of biofilms formed by wild-type, mmpL11 mutant, and complemented strains.

Techniques Used: Mutagenesis, Incubation, Electron Microscopy

18) Product Images from "Antigen 84, an Effector of Pleiomorphism in Mycobacterium smegmatis ▿"

Article Title: Antigen 84, an Effector of Pleiomorphism in Mycobacterium smegmatis ▿

Journal:

doi: 10.1128/JB.00726-07

Oligomerization of mycobacterial antigen 84. (A) Subcellular localization of Ag84 in M. bovis BCG lysates. Bacterial cultures were grown at 37°C in 7H9-OADC medium, and cellular fractionation was carried out as described in Materials and Methods.
Figure Legend Snippet: Oligomerization of mycobacterial antigen 84. (A) Subcellular localization of Ag84 in M. bovis BCG lysates. Bacterial cultures were grown at 37°C in 7H9-OADC medium, and cellular fractionation was carried out as described in Materials and Methods.

Techniques Used: Cell Fractionation

19) Product Images from "Mycobacterium tuberculosis Pst/SenX3-RegX3 Regulates Membrane Vesicle Production Independently of ESX-5 Activity"

Article Title: Mycobacterium tuberculosis Pst/SenX3-RegX3 Regulates Membrane Vesicle Production Independently of ESX-5 Activity

Journal: mBio

doi: 10.1128/mBio.00778-18

Incomplete repression of eccD 5 transcription has moderate effects on growth. (A to D) Wild-type M. tuberculosis Erdman (WT) and the Δ pstA1 , eccD 5 Tet-OFF, and Δ pstA1 eccD 5 Tet-OFF strains were inoculated in 7H9 complete medium at an OD 600 of 0.05 and grown at 37°C with aeration. Anhydrotetracycline hydrochloride (ATc; 100 ng/ml) was added at day 0 and day 7 as indicated. Growth was monitored by daily OD 600 measurements (A and C) and by plating serially diluted cultures on 7H10 medium to determine numbers of viable CFU per milliliter on days 0, 3, 6, 9, 12, and 15 (B and D). The key in panel B applies to panels A and B; the key in panel D applies to panels C and D. **, P
Figure Legend Snippet: Incomplete repression of eccD 5 transcription has moderate effects on growth. (A to D) Wild-type M. tuberculosis Erdman (WT) and the Δ pstA1 , eccD 5 Tet-OFF, and Δ pstA1 eccD 5 Tet-OFF strains were inoculated in 7H9 complete medium at an OD 600 of 0.05 and grown at 37°C with aeration. Anhydrotetracycline hydrochloride (ATc; 100 ng/ml) was added at day 0 and day 7 as indicated. Growth was monitored by daily OD 600 measurements (A and C) and by plating serially diluted cultures on 7H10 medium to determine numbers of viable CFU per milliliter on days 0, 3, 6, 9, 12, and 15 (B and D). The key in panel B applies to panels A and B; the key in panel D applies to panels C and D. **, P

Techniques Used:

20) Product Images from "A rheostat mechanism governs the bifurcation of carbon flux in mycobacteria"

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

Journal: Nature Communications

doi: 10.1038/ncomms12527

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).
Figure Legend 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).

Techniques Used: Concentration Assay, Transferring

Related Articles

Clone Assay:

Article Title: Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro
Article Snippet: The KatG-overexpressing construct consists of a 2.9-kb Eco RI fragment carrying the katG gene of H37Rv cloned into the polylinker site of pMV306 ( ). .. Mycobacterial strains were grown for 7 days in Middlebrook 7H9 liquid medium (Difco, Detroit, Mich.) containing 0.05% Tween 80 (Sigma Chemical Co., St. Louis, Mo.) at 37°C with daily agitation.

Article Title: Mycobacterial Gene cuvA Is Required for Optimal Nutrient Utilization and Virulence
Article Snippet: Escherichia coli TOP10 (Invitrogen) was used for cloning and was grown in LB broth. .. For routine growth, M. tuberculosis and M. smegmatis were grown at 37°C in Middlebrook 7H9 liquid medium (Difco) supplemented with 0.5% albumin, 0.2% glucose, 0.085% NaCl, 0.2% glycerol, and 0.05% Tween 80 (7H9-ADC-Tw).

Amplification:

Article Title: A rheostat mechanism governs the bifurcation of carbon flux in mycobacteria
Article Snippet: .. Individual colonies were picked and amplified in 7H9 liquid medium (no antibiotics) and then plated on LB agar or 7H10 containing 5% sucrose to select for cells in which plasmid excision had occurred (second crossover). .. Individual colonies were picked and the replacement of the gene by the fusion gene or gene deletion was confirmed by PCR.

Binding Assay:

Article Title: Mycobacterium tuberculosis Requires Regulation of ESX-5 Secretion for Virulence in Irgm1-Deficient Mice
Article Snippet: Construction of strains harboring mutations in the esx-5 RegX3 binding site sequence is described below. .. Bacterial cultures were grown at 37°C with aeration in Middlebrook 7H9 liquid medium (Difco) supplemented with albumin-dextrose-saline (ADS), 0.5% glycerol, and 0.1% Tween 80 or on Middlebrook 7H10 agar medium (Difco) supplemented with 10% Middlebrook oleic acid-albumin-dextrose-catalase (OADC) (BD Biosciences) and 0.5% glycerol, unless otherwise noted.

Southern Blot:

Article Title: Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro
Article Snippet: The strains used include the laboratory strains H37Rv (Trudeau Institute, Saranac Lake, N.Y.), CDC 1551 (T. M. Shinnick, Centers for Disease Control and Prevention, Atlanta, Ga.), ATCC 35825, and H37Rv Inhr , which was selected from H37Rv by direct plating onto 10 μg of isoniazid per ml (loss of KatG was verified by [i] activity assay, [ii] Western blotting using anti-KatG antipeptide antisera, and [iii] Southern blot analysis using the katG probe and restriction digestion as described previously [ ]), as well as the recombinant strain H37Rv(pMH59) and the recombinant strain H37Rv(pMH91), which have been previously described ( ). .. Mycobacterial strains were grown for 7 days in Middlebrook 7H9 liquid medium (Difco, Detroit, Mich.) containing 0.05% Tween 80 (Sigma Chemical Co., St. Louis, Mo.) at 37°C with daily agitation.

Mutagenesis:

Article Title: Phosphate Starvation: a Novel Signal that Triggers ESX-5 Secretion in Mycobacterium tuberculosis
Article Snippet: The Δ pstA1 Δ ppe27-pe19 , Δ pstA1 Δ ppe27 , and Δ pstA1 Δ pe19 mutant strains were constructed as described ( ). .. Bacterial cultures were grown at 37°C with aeration in Middlebrook 7H9 liquid medium (Difco) supplemented with albumin-dextrose-saline (ADS), 0.5% glycerol and 0.1% Tween-80, unless otherwise noted.

Article Title: Mycobacterium tuberculosis Requires Regulation of ESX-5 Secretion for Virulence in Irgm1-Deficient Mice
Article Snippet: M. tuberculosis Erdman and the derivative Δ pstA1 , Δ regX3 , and Δ pstA1 Δ regX3 mutant strains were previously described ( ). .. Bacterial cultures were grown at 37°C with aeration in Middlebrook 7H9 liquid medium (Difco) supplemented with albumin-dextrose-saline (ADS), 0.5% glycerol, and 0.1% Tween 80 or on Middlebrook 7H10 agar medium (Difco) supplemented with 10% Middlebrook oleic acid-albumin-dextrose-catalase (OADC) (BD Biosciences) and 0.5% glycerol, unless otherwise noted.

Isolation:

Article Title: Phosphate Starvation: a Novel Signal that Triggers ESX-5 Secretion in Mycobacterium tuberculosis
Article Snippet: Bacterial cultures were grown at 37°C with aeration in Middlebrook 7H9 liquid medium (Difco) supplemented with albumin-dextrose-saline (ADS), 0.5% glycerol and 0.1% Tween-80, unless otherwise noted. .. Sauton's medium (3.67 mM KH2 PO4 , 2 mM MgSO4 -7H2 O, 9.5 mM citric acid, 0.19 mM ammonium iron (III) citrate, 26.64 mM L-asparagine, 6% glycerol, 0.01% ZnSO4 , pH 7.4) was used to grow cultures for protein isolation.

Article Title: Chlorine Disinfection of Atypical Mycobacteria Isolated from a Water Distribution System
Article Snippet: The other mycobacterial strains used were isolated from cold public water supplies in Paris. .. Cells were grown in Middlebrook 7H9 liquid medium (Difco Laboratories, West Molesley, Surrey, United Kingdom) containing 10% (vol/vol) oleic acid-albumin enrichment and 0.05% (vol/vol) Tween 80 at their optimal growth temperature (30 or 37°C, depending on the strain) in a rotary shaker (120 rpm).

Article Title: Mycobacterium tuberculosis Requires Regulation of ESX-5 Secretion for Virulence in Irgm1-Deficient Mice
Article Snippet: Bacterial cultures were grown at 37°C with aeration in Middlebrook 7H9 liquid medium (Difco) supplemented with albumin-dextrose-saline (ADS), 0.5% glycerol, and 0.1% Tween 80 or on Middlebrook 7H10 agar medium (Difco) supplemented with 10% Middlebrook oleic acid-albumin-dextrose-catalase (OADC) (BD Biosciences) and 0.5% glycerol, unless otherwise noted. .. Sauton’s medium [3.67 mM KH2 PO4 , 2 mM MgSO4 ·7H2 O, 9.5 mM citric acid, 0.19 mM ammonium iron(III) citrate, 26.64 mM l -asparagine, 6% glycerol, 0.01% ZnSO4 , pH 7.4] or Pi -limited Sauton’s medium (Sauton’s containing 2.5 μM KH2 PO4 and buffered with 50 mM morpholinepropanesulfonic acid [MOPS], pH 7.4) were used to grow cultures for protein isolation.

Cell Culture:

Article Title: The Mycobacterium tuberculosis MmpL11 Cell Wall Lipid Transporter Is Important for Biofilm Formation, Intracellular Growth, and Nonreplicating Persistence
Article Snippet: Mycobacterial strains were routinely maintained in Middlebrook 7H9 liquid medium (Difco) or on Middlebrook 7H11 agar (Difco) plates supplemented with 10% oleic acid-albumin-dextrose-catalase (OADC; Difco) or albumin dextrose salts (ADS) containing 8.1 mg ml−1 NaCl, 50 mg ml−1 bovine serum albumin (BSA), and 20 mg ml−1 dextrose. .. Where indicated, bacteria were cultured in Sauton's medium containing 0.5 g liter−1 K2 HPO4 , 0.5 g liter−1 MgSO4 , 4.0 g liter−1 l -asparagine, 0.05 g liter−1 ferric ammonium citrate, 4.76% glycerol, and 1.0 mg liter−1 ZnSO4 , with a final pH of 7.0.

Concentration Assay:

Article Title: The Mycobacterium tuberculosis MmpL11 Cell Wall Lipid Transporter Is Important for Biofilm Formation, Intracellular Growth, and Nonreplicating Persistence
Article Snippet: Mycobacterial strains were routinely maintained in Middlebrook 7H9 liquid medium (Difco) or on Middlebrook 7H11 agar (Difco) plates supplemented with 10% oleic acid-albumin-dextrose-catalase (OADC; Difco) or albumin dextrose salts (ADS) containing 8.1 mg ml−1 NaCl, 50 mg ml−1 bovine serum albumin (BSA), and 20 mg ml−1 dextrose. .. Glycerol was added to 7H9 and 7H11 media at a final concentration of 0.5%.

Article Title: Mycobacterium tuberculosis Requires Regulation of ESX-5 Secretion for Virulence in Irgm1-Deficient Mice
Article Snippet: Bacterial cultures were grown at 37°C with aeration in Middlebrook 7H9 liquid medium (Difco) supplemented with albumin-dextrose-saline (ADS), 0.5% glycerol, and 0.1% Tween 80 or on Middlebrook 7H10 agar medium (Difco) supplemented with 10% Middlebrook oleic acid-albumin-dextrose-catalase (OADC) (BD Biosciences) and 0.5% glycerol, unless otherwise noted. .. Frozen stocks were prepared by growing liquid cultures to mid-exponential phase (optical density at 600 nm [OD600 ], 0.8 to 1.0) in complete 7H9 medium, adding glycerol to a 15% final concentration, and storing 1-ml aliquots at −80°C.

Incubation:

Article Title: MmpL11 Protein Transports Mycolic Acid-containing Lipids to the Mycobacterial Cell Wall and Contributes to Biofilm Formation in Mycobacterium smegmatis *
Article Snippet: Mycobacterial strains were maintained in Middlebrook 7H9 liquid medium (Difco) or on Middlebrook 7H11 agar plates (Difco) supplemented with 10% albumin/dextrose/saline. .. When required, cultures were incubated with the antibiotics kanamycin (25 μg/ml) and hygromycin (75 μg/ml).

Diffusion-based Assay:

Article Title: Octahedral ruthenium (II) polypyridyl complexes as antimicrobial agents against mycobacterium
Article Snippet: Disc diffusion assays were performed according to CLSI guidelines. .. Bacteria were grown in TSB medium; C. neoformans and C. albicans were grown in Yeast Extract Peptone Dextrose (YPD) medium; M. smegmatis mc2 155 was grown in 7H9 liquid medium (Difco) supplemented with 0.05% w/v Tween 80, 0.5% glycerol and 0.5% glucose or were grown on 7H10 agar supplemented with 1% glycerol and 0.5% glucose.

Activity Assay:

Article Title: Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro
Article Snippet: The strains used include the laboratory strains H37Rv (Trudeau Institute, Saranac Lake, N.Y.), CDC 1551 (T. M. Shinnick, Centers for Disease Control and Prevention, Atlanta, Ga.), ATCC 35825, and H37Rv Inhr , which was selected from H37Rv by direct plating onto 10 μg of isoniazid per ml (loss of KatG was verified by [i] activity assay, [ii] Western blotting using anti-KatG antipeptide antisera, and [iii] Southern blot analysis using the katG probe and restriction digestion as described previously [ ]), as well as the recombinant strain H37Rv(pMH59) and the recombinant strain H37Rv(pMH91), which have been previously described ( ). .. Mycobacterial strains were grown for 7 days in Middlebrook 7H9 liquid medium (Difco, Detroit, Mich.) containing 0.05% Tween 80 (Sigma Chemical Co., St. Louis, Mo.) at 37°C with daily agitation.

Construct:

Article Title: Phosphate Starvation: a Novel Signal that Triggers ESX-5 Secretion in Mycobacterium tuberculosis
Article Snippet: The Δ pstA1 Δ ppe27-pe19 , Δ pstA1 Δ ppe27 , and Δ pstA1 Δ pe19 mutant strains were constructed as described ( ). .. Bacterial cultures were grown at 37°C with aeration in Middlebrook 7H9 liquid medium (Difco) supplemented with albumin-dextrose-saline (ADS), 0.5% glycerol and 0.1% Tween-80, unless otherwise noted.

Article Title: Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro
Article Snippet: This construct contains about 85 bp upstream of the GTG start codon of katG . .. Mycobacterial strains were grown for 7 days in Middlebrook 7H9 liquid medium (Difco, Detroit, Mich.) containing 0.05% Tween 80 (Sigma Chemical Co., St. Louis, Mo.) at 37°C with daily agitation.

Expressing:

Article Title: Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro
Article Snippet: The vector system also has a synthetic promoter element incorporated into the polylinker driving expression of KatG ( ). .. Mycobacterial strains were grown for 7 days in Middlebrook 7H9 liquid medium (Difco, Detroit, Mich.) containing 0.05% Tween 80 (Sigma Chemical Co., St. Louis, Mo.) at 37°C with daily agitation.

Western Blot:

Article Title: Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro
Article Snippet: The strains used include the laboratory strains H37Rv (Trudeau Institute, Saranac Lake, N.Y.), CDC 1551 (T. M. Shinnick, Centers for Disease Control and Prevention, Atlanta, Ga.), ATCC 35825, and H37Rv Inhr , which was selected from H37Rv by direct plating onto 10 μg of isoniazid per ml (loss of KatG was verified by [i] activity assay, [ii] Western blotting using anti-KatG antipeptide antisera, and [iii] Southern blot analysis using the katG probe and restriction digestion as described previously [ ]), as well as the recombinant strain H37Rv(pMH59) and the recombinant strain H37Rv(pMH91), which have been previously described ( ). .. Mycobacterial strains were grown for 7 days in Middlebrook 7H9 liquid medium (Difco, Detroit, Mich.) containing 0.05% Tween 80 (Sigma Chemical Co., St. Louis, Mo.) at 37°C with daily agitation.

Sequencing:

Article Title: Mycobacterium tuberculosis Requires Regulation of ESX-5 Secretion for Virulence in Irgm1-Deficient Mice
Article Snippet: Construction of strains harboring mutations in the esx-5 RegX3 binding site sequence is described below. .. Bacterial cultures were grown at 37°C with aeration in Middlebrook 7H9 liquid medium (Difco) supplemented with albumin-dextrose-saline (ADS), 0.5% glycerol, and 0.1% Tween 80 or on Middlebrook 7H10 agar medium (Difco) supplemented with 10% Middlebrook oleic acid-albumin-dextrose-catalase (OADC) (BD Biosciences) and 0.5% glycerol, unless otherwise noted.

Lysis:

Article Title: Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro
Article Snippet: Mycobacterial strains were grown for 7 days in Middlebrook 7H9 liquid medium (Difco, Detroit, Mich.) containing 0.05% Tween 80 (Sigma Chemical Co., St. Louis, Mo.) at 37°C with daily agitation. .. The culture medium was lipopolysaccharide free and without reactivity in the Limulus amoebocyte lysis assay (Whittaker Bioproducts, Walkersville, Md.).

Recombinant:

Article Title: Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro
Article Snippet: The strains used include the laboratory strains H37Rv (Trudeau Institute, Saranac Lake, N.Y.), CDC 1551 (T. M. Shinnick, Centers for Disease Control and Prevention, Atlanta, Ga.), ATCC 35825, and H37Rv Inhr , which was selected from H37Rv by direct plating onto 10 μg of isoniazid per ml (loss of KatG was verified by [i] activity assay, [ii] Western blotting using anti-KatG antipeptide antisera, and [iii] Southern blot analysis using the katG probe and restriction digestion as described previously [ ]), as well as the recombinant strain H37Rv(pMH59) and the recombinant strain H37Rv(pMH91), which have been previously described ( ). .. Mycobacterial strains were grown for 7 days in Middlebrook 7H9 liquid medium (Difco, Detroit, Mich.) containing 0.05% Tween 80 (Sigma Chemical Co., St. Louis, Mo.) at 37°C with daily agitation.

Modification:

Article Title: MmpL11 Protein Transports Mycolic Acid-containing Lipids to the Mycobacterial Cell Wall and Contributes to Biofilm Formation in Mycobacterium smegmatis *
Article Snippet: Mycobacterial strains were maintained in Middlebrook 7H9 liquid medium (Difco) or on Middlebrook 7H11 agar plates (Difco) supplemented with 10% albumin/dextrose/saline. .. For biofilm growth, M. smegmatis was grown in polystyrene Petri dishes at 30 °C in modified Sauton's medium without Tween 80.

other:

Article Title: The Mycobacterium tuberculosis relBE toxin:antitoxin genes are stress-responsive modules that regulate growth through translation inhibition
Article Snippet: M. smegmatis mc2 155 was grown with aeration at 37°C in Middlebrook 7H9 liquid medium (Difco) supplemented with 0.05% Tw or on Luria-Bertani (LB) agar plates ( ).

Article Title: Nitrogen starvation-induced transcriptome alterations and influence of transcription regulator mutants in Mycobacterium smegmatis
Article Snippet: Mycobacterial strains were grown in Middlebrook 7H9 liquid medium (Difco Laboratories; per 900 ml approx.

Article Title: In Vitro Model of Mycobacterial Growth Arrest Using Nitric Oxide with Limited Air ▿
Article Snippet: M. bovis BCG, substrain Pasteur, isolate KD1295 , was grown in Middlebrook 7H9 liquid medium (catalog no. 271310; Difco) or on 7H10 agar medium (catalog no. 262710; Difco) supplemented with 10% albumin dextrose complex, 0.2% glycerol, and 0.05% Tween 80 ( ).

Plasmid Preparation:

Article Title: Mycobacterium tuberculosis Catalase and Peroxidase Activities and Resistance to Oxidative Killing in Human Monocytes In Vitro
Article Snippet: The vector system also has a synthetic promoter element incorporated into the polylinker driving expression of KatG ( ). .. Mycobacterial strains were grown for 7 days in Middlebrook 7H9 liquid medium (Difco, Detroit, Mich.) containing 0.05% Tween 80 (Sigma Chemical Co., St. Louis, Mo.) at 37°C with daily agitation.

Article Title: A rheostat mechanism governs the bifurcation of carbon flux in mycobacteria
Article Snippet: .. Individual colonies were picked and amplified in 7H9 liquid medium (no antibiotics) and then plated on LB agar or 7H10 containing 5% sucrose to select for cells in which plasmid excision had occurred (second crossover). .. Individual colonies were picked and the replacement of the gene by the fusion gene or gene deletion was confirmed by PCR.

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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 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/7h9 liquid medium/product/Difco
    Average 94 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    7h9 liquid medium - by Bioz Stars, 2020-04
    94/100 stars
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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 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h9 medium/product/Difco
    Average 95 stars, based on 36 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h9 medium - by Bioz Stars, 2020-04
    95/100 stars
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    94
    Difco middlebrook 7h9
    Toxic effect of the five toxins on the growth of M. smegmatis . (A) Transformation of pMV306AC-toxin plasmid into M. smegmatis mc 2 155. One microgram of DNA was electrophorated and cells were plated on <t>Middlebrook</t> 7H10 solid media. The plates were incubated for 3 days at 37°C. (B) Effect of toxin on the induction on colony formation of M. smegmatis mc 2 155. Each transformant from (A) was grown for 3 days at 37°C. The culture was diluted 100-fold into Middlebrook <t>7H9</t> liquid media and streaked on Middlebrook 7H10 solid media in the absence and presence of 0.2% acetamide. The numbers in the circle correspond to those in (A) .
    Middlebrook 7h9, supplied by Difco, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h9/product/Difco
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h9 - by Bioz Stars, 2020-04
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    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

    Effect of DETA-NO on growth of cultured M. bovis BCG. Bacteria growing exponentially in 7H9 medium were transferred to evacuated (Vacutainer) tubes at mid-log phase, and at time zero either the cells were left untreated (open circles) or DETA-NO was added

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: In Vitro Model of Mycobacterial Growth Arrest Using Nitric Oxide with Limited Air ▿

    doi: 10.1128/AAC.00442-08

    Figure Lengend Snippet: Effect of DETA-NO on growth of cultured M. bovis BCG. Bacteria growing exponentially in 7H9 medium were transferred to evacuated (Vacutainer) tubes at mid-log phase, and at time zero either the cells were left untreated (open circles) or DETA-NO was added

    Article Snippet: M. bovis BCG, substrain Pasteur, isolate KD1295 , was grown in Middlebrook 7H9 liquid medium (catalog no. 271310; Difco) or on 7H10 agar medium (catalog no. 262710; Difco) supplemented with 10% albumin dextrose complex, 0.2% glycerol, and 0.05% Tween 80 ( ).

    Techniques: Cell Culture

    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:

    Toxic effect of the five toxins on the growth of M. smegmatis . (A) Transformation of pMV306AC-toxin plasmid into M. smegmatis mc 2 155. One microgram of DNA was electrophorated and cells were plated on Middlebrook 7H10 solid media. The plates were incubated for 3 days at 37°C. (B) Effect of toxin on the induction on colony formation of M. smegmatis mc 2 155. Each transformant from (A) was grown for 3 days at 37°C. The culture was diluted 100-fold into Middlebrook 7H9 liquid media and streaked on Middlebrook 7H10 solid media in the absence and presence of 0.2% acetamide. The numbers in the circle correspond to those in (A) .

    Journal: Frontiers in Microbiology

    Article Title: Functional Studies of Five Toxin-Antitoxin Modules in Mycobacterium tuberculosis H37Rv

    doi: 10.3389/fmicb.2016.02071

    Figure Lengend Snippet: Toxic effect of the five toxins on the growth of M. smegmatis . (A) Transformation of pMV306AC-toxin plasmid into M. smegmatis mc 2 155. One microgram of DNA was electrophorated and cells were plated on Middlebrook 7H10 solid media. The plates were incubated for 3 days at 37°C. (B) Effect of toxin on the induction on colony formation of M. smegmatis mc 2 155. Each transformant from (A) was grown for 3 days at 37°C. The culture was diluted 100-fold into Middlebrook 7H9 liquid media and streaked on Middlebrook 7H10 solid media in the absence and presence of 0.2% acetamide. The numbers in the circle correspond to those in (A) .

    Article Snippet: M. smegmatis mc2 155 was grown in Middlebrook 7H9 (a liquid medium, Difco) containing 0.2% glycerol and 0.02% Tween 80 or Middlebrook 7H10 (a solid agar medium, Difco) containing 0.2% glycerol.

    Techniques: Transformation Assay, Plasmid Preparation, Incubation