a baumannii atcc 17978  (ATCC)


Bioz Verified Symbol ATCC is a verified supplier
Bioz Manufacturer Symbol ATCC manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Acinetobacter baumannii 5377 NCDC KC 755
    Description:

    Catalog Number:
    17978
    Price:
    None
    Buy from Supplier


    Structured Review

    ATCC a baumannii atcc 17978
    The effect of Zn and Cu treatment on the resistance of A. <t>baumannii</t> to oxidative stress. The effect of ( a ) 40 μM paraquat and ( e ) 160 μM H 2 O 2 on A. baumannii strain <t>ATCC</t> 17978 grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions, was determined by measuring the optical density at 600 nm (OD 600 ). The growth delay between untreated (UT) and paraquat- or H 2 O 2 -treated cells was examined by comparing the EC 50 under each relevant metal ion stress condition. Statistical analyses were performed using a one-way ANOVA. b The total SOD activity of mid-log phase cells (untreated, 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu) was determined. Data were corrected for total protein content, followed by normalisation to untreated cells. Statistical analyses were performed using a Student’s t -test. The mRNA transcription levels of ( c ) Cu-Zn superoxide dismutase (Cu-Zn-SOD; A1S_3143), ( d ) Fe/Mn superoxide dismutase (Fe/Mn-SOD; A1S_2343) were determined by qRT-PCR. The transcription levels examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu were corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest. For all statistical analyses; ns = not significant, * = p

    https://www.bioz.com/result/a baumannii atcc 17978/product/ATCC
    Average 99 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    a baumannii atcc 17978 - by Bioz Stars, 2020-08
    99/100 stars

    Images

    1) Product Images from "Zinc stress induces copper depletion in Acinetobacter baumannii"

    Article Title: Zinc stress induces copper depletion in Acinetobacter baumannii

    Journal: BMC Microbiology

    doi: 10.1186/s12866-017-0965-y

    The effect of Zn and Cu treatment on the resistance of A. baumannii to oxidative stress. The effect of ( a ) 40 μM paraquat and ( e ) 160 μM H 2 O 2 on A. baumannii strain ATCC 17978 grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions, was determined by measuring the optical density at 600 nm (OD 600 ). The growth delay between untreated (UT) and paraquat- or H 2 O 2 -treated cells was examined by comparing the EC 50 under each relevant metal ion stress condition. Statistical analyses were performed using a one-way ANOVA. b The total SOD activity of mid-log phase cells (untreated, 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu) was determined. Data were corrected for total protein content, followed by normalisation to untreated cells. Statistical analyses were performed using a Student’s t -test. The mRNA transcription levels of ( c ) Cu-Zn superoxide dismutase (Cu-Zn-SOD; A1S_3143), ( d ) Fe/Mn superoxide dismutase (Fe/Mn-SOD; A1S_2343) were determined by qRT-PCR. The transcription levels examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu were corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest. For all statistical analyses; ns = not significant, * = p
    Figure Legend Snippet: The effect of Zn and Cu treatment on the resistance of A. baumannii to oxidative stress. The effect of ( a ) 40 μM paraquat and ( e ) 160 μM H 2 O 2 on A. baumannii strain ATCC 17978 grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions, was determined by measuring the optical density at 600 nm (OD 600 ). The growth delay between untreated (UT) and paraquat- or H 2 O 2 -treated cells was examined by comparing the EC 50 under each relevant metal ion stress condition. Statistical analyses were performed using a one-way ANOVA. b The total SOD activity of mid-log phase cells (untreated, 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu) was determined. Data were corrected for total protein content, followed by normalisation to untreated cells. Statistical analyses were performed using a Student’s t -test. The mRNA transcription levels of ( c ) Cu-Zn superoxide dismutase (Cu-Zn-SOD; A1S_3143), ( d ) Fe/Mn superoxide dismutase (Fe/Mn-SOD; A1S_2343) were determined by qRT-PCR. The transcription levels examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu were corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest. For all statistical analyses; ns = not significant, * = p

    Techniques Used: Activity Assay, Quantitative RT-PCR

    An overview of the cellular responses of A. baumannii to Zn and Cu stress. Cartoon representation of A. baumannii ATCC 17978 under either high Zn (400 μM), high Cu (400 μM) or high Zn + Cu (400 μM each) conditions based on the observations from this work. The numbers within the efflux systems (ovals) are the locus-tags for A. baumannii strain ATCC 17978, excluding their prefix “A1S_”. The efflux proteins in red are those up-regulated under high Zn, in blue by high Cu and those in purple were found to be transcriptionally responsive to both high Zn and Cu independently (A1S_2939 and A1S_0709) or the combination of high Zn and high Cu (A1S_2939 and A1S_3098). Under high Zn and high Zn + Cu conditions, A. baumannii becomes increasingly sensitive to superoxide stress (O 2 .- in the red burst), but shows increased resistance to hydrogen peroxide (green cloud). Under high Cu conditions, A. baumannii shows increased susceptibility to hydrogen peroxide (H 2 O 2 in the red burst). Under high Zn conditions and high Zn + Cu conditions, Cu levels reduce to 74 and 62%, respectively
    Figure Legend Snippet: An overview of the cellular responses of A. baumannii to Zn and Cu stress. Cartoon representation of A. baumannii ATCC 17978 under either high Zn (400 μM), high Cu (400 μM) or high Zn + Cu (400 μM each) conditions based on the observations from this work. The numbers within the efflux systems (ovals) are the locus-tags for A. baumannii strain ATCC 17978, excluding their prefix “A1S_”. The efflux proteins in red are those up-regulated under high Zn, in blue by high Cu and those in purple were found to be transcriptionally responsive to both high Zn and Cu independently (A1S_2939 and A1S_0709) or the combination of high Zn and high Cu (A1S_2939 and A1S_3098). Under high Zn and high Zn + Cu conditions, A. baumannii becomes increasingly sensitive to superoxide stress (O 2 .- in the red burst), but shows increased resistance to hydrogen peroxide (green cloud). Under high Cu conditions, A. baumannii shows increased susceptibility to hydrogen peroxide (H 2 O 2 in the red burst). Under high Zn conditions and high Zn + Cu conditions, Cu levels reduce to 74 and 62%, respectively

    Techniques Used:

    The effects of Zn and Cu on A. baumannii membrane biology. The major cellular fatty acid constituents of A. baumannii strain ATCC 17978, grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions were determined by gas chromatography. The abundance of fatty acids, expressed as percentage of total cellular fatty acids, is ( a ) 16:0 ( b ) 18:0, ( c ) 16:1n-7 and ( d ) 18:1n-7. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, ** = p
    Figure Legend Snippet: The effects of Zn and Cu on A. baumannii membrane biology. The major cellular fatty acid constituents of A. baumannii strain ATCC 17978, grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions were determined by gas chromatography. The abundance of fatty acids, expressed as percentage of total cellular fatty acids, is ( a ) 16:0 ( b ) 18:0, ( c ) 16:1n-7 and ( d ) 18:1n-7. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, ** = p

    Techniques Used: Gas Chromatography

    The effect of Zn and Cu on macrophage-mediated killing of A. baumannii . The survival of A. baumannii ATCC 17978 cells in the presence of THP-1 human monocyte-derived macrophages that were either treated with 50 μM Zn, 50 μM Cu or 50 μM Zn + 50 μM Cu was compared to survival of A. baumannii ATCC 17978 cells in the presence of untreated THP-1 cells. The data are the mean of biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant, * = p
    Figure Legend Snippet: The effect of Zn and Cu on macrophage-mediated killing of A. baumannii . The survival of A. baumannii ATCC 17978 cells in the presence of THP-1 human monocyte-derived macrophages that were either treated with 50 μM Zn, 50 μM Cu or 50 μM Zn + 50 μM Cu was compared to survival of A. baumannii ATCC 17978 cells in the presence of untreated THP-1 cells. The data are the mean of biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant, * = p

    Techniques Used: Derivative Assay, Two Tailed Test

    The effect of Zn and Cu stress on A. baumannii growth and metal ion homeostasis. Growth as determined by measuring the optical density at 600 nm (OD 600 ) of A. baumannii strain ATCC 17978 under increasing concentrations of ( a ) Zn, ( b ) Cu, or ( c ) Zn + Cu ( n ≥ 3). Examination of the ( d ) Zn, ( e ) Cu and ( f ) Fe levels in untreated cells and cells grown in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu. The cellular metal ion content (in microgram) per gram of desiccated cells was determined by ICP-MS. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant and * = p
    Figure Legend Snippet: The effect of Zn and Cu stress on A. baumannii growth and metal ion homeostasis. Growth as determined by measuring the optical density at 600 nm (OD 600 ) of A. baumannii strain ATCC 17978 under increasing concentrations of ( a ) Zn, ( b ) Cu, or ( c ) Zn + Cu ( n ≥ 3). Examination of the ( d ) Zn, ( e ) Cu and ( f ) Fe levels in untreated cells and cells grown in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu. The cellular metal ion content (in microgram) per gram of desiccated cells was determined by ICP-MS. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant and * = p

    Techniques Used: Mass Spectrometry, Two Tailed Test

    Transcriptional analyses of putative A. baumannii Zn and Cu efflux systems. The mRNA transcription levels of the putative ( a ) Cation Diffusion Facilitator (CDF), ( b ) Heavy Metal Efflux (HME), ( c ) P-type ATPase, ( d ) CorA-type and ( e ) CopB-like copper resistance genes of A. baumannii strain ATCC 17978 were determined by qRT-PCR. Transcription levels were examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu and corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, * = p
    Figure Legend Snippet: Transcriptional analyses of putative A. baumannii Zn and Cu efflux systems. The mRNA transcription levels of the putative ( a ) Cation Diffusion Facilitator (CDF), ( b ) Heavy Metal Efflux (HME), ( c ) P-type ATPase, ( d ) CorA-type and ( e ) CopB-like copper resistance genes of A. baumannii strain ATCC 17978 were determined by qRT-PCR. Transcription levels were examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu and corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, * = p

    Techniques Used: Diffusion-based Assay, Quantitative RT-PCR

    2) Product Images from "Identification of genes essential for pellicle formation in Acinetobacter baumannii"

    Article Title: Identification of genes essential for pellicle formation in Acinetobacter baumannii

    Journal: BMC Microbiology

    doi: 10.1186/s12866-015-0440-6

    Motility of A. baumannii ATCC 17978, 17978hm, and transposon insertion mutants cpdA ::Tn, A1S_0112::Tn and A1S_0115::Tn. Overnight grown bacterial cells were used to inoculate Luria-Bertani media containing 0.25 % agar; plates were grown overnight at 37 °C. The A1S_0115::Tn, A1S_0115(0115) and A1S_0115(1266) transposon mutant strains and the A1S_0112::Tn, A1S_0112::Tn(0112) and A1S_0112::Tn(1266) transposon mutant strains only show growth at the initial inoculum site at the centre of the plate giving a non-motile phenotype. However, the cpdA ::Tn, cpdA ::Tn(0249) and cpdA ::Tn(1266) transposon mutant strains retained motility. The complemented strain cpdA ::Tn(0249) displayed a motility pattern almost identical to the 17978hm parent, whereas cpdA ::Tn and cpdA ::Tn(1266) displayed a “spoke-like” motility pattern. ATCC 17978 is included as a control displaying a non-motile phenotype
    Figure Legend Snippet: Motility of A. baumannii ATCC 17978, 17978hm, and transposon insertion mutants cpdA ::Tn, A1S_0112::Tn and A1S_0115::Tn. Overnight grown bacterial cells were used to inoculate Luria-Bertani media containing 0.25 % agar; plates were grown overnight at 37 °C. The A1S_0115::Tn, A1S_0115(0115) and A1S_0115(1266) transposon mutant strains and the A1S_0112::Tn, A1S_0112::Tn(0112) and A1S_0112::Tn(1266) transposon mutant strains only show growth at the initial inoculum site at the centre of the plate giving a non-motile phenotype. However, the cpdA ::Tn, cpdA ::Tn(0249) and cpdA ::Tn(1266) transposon mutant strains retained motility. The complemented strain cpdA ::Tn(0249) displayed a motility pattern almost identical to the 17978hm parent, whereas cpdA ::Tn and cpdA ::Tn(1266) displayed a “spoke-like” motility pattern. ATCC 17978 is included as a control displaying a non-motile phenotype

    Techniques Used: Mutagenesis

    3) Product Images from "Identification of a General O-linked Protein Glycosylation System in Acinetobacter baumannii and Its Role in Virulence and Biofilm Formation"

    Article Title: Identification of a General O-linked Protein Glycosylation System in Acinetobacter baumannii and Its Role in Virulence and Biofilm Formation

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1002758

    Identification of additional glycoproteins in A. baumannii ATCC 17978. Tryptically digested membranes were enriched via ZIC-HILIC and analyzed by LC-MS and HCD MS-MS. All spectra were analyzed for the diagnostic oxonium ion of 301.10 m/z, and positive spectra were analyzed manually to identify the glycopeptide. This spectra is representative of each glycopeptide identified in Table 2 . A) ITMS-CID of the precursor ion at m/z 1999.943 reveals the pentasaccharide attached to the peptide AKPASTPAVK . B) FTMS-HCD of the precursor ion at m/z 1999.943 reveals the peptide sequence AKPASTPAVK .
    Figure Legend Snippet: Identification of additional glycoproteins in A. baumannii ATCC 17978. Tryptically digested membranes were enriched via ZIC-HILIC and analyzed by LC-MS and HCD MS-MS. All spectra were analyzed for the diagnostic oxonium ion of 301.10 m/z, and positive spectra were analyzed manually to identify the glycopeptide. This spectra is representative of each glycopeptide identified in Table 2 . A) ITMS-CID of the precursor ion at m/z 1999.943 reveals the pentasaccharide attached to the peptide AKPASTPAVK . B) FTMS-HCD of the precursor ion at m/z 1999.943 reveals the peptide sequence AKPASTPAVK .

    Techniques Used: Hydrophilic Interaction Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Diagnostic Assay, Sequencing

    Characterization of A. baumannii ATCC 17978 pathogenesis in a murine septicemia model. A) Determination of the LD 50 of A. baumannii ATCC 17978. Groups of 5 mice were injected with serial dilutions of A. baumannii WT to determine the LD 50 which was calculated to be 6.49×10 4 CFU @ 18 hrs post infection. B) Murine competition septicemia between A. baumannii WT and ΔpglL . Groups of 3 mice were injected with ∼1∶1 WT to Δ pglL CFU's and were sacrificed after 18 hrs, spleens were harvested, and bacterial load determined.
    Figure Legend Snippet: Characterization of A. baumannii ATCC 17978 pathogenesis in a murine septicemia model. A) Determination of the LD 50 of A. baumannii ATCC 17978. Groups of 5 mice were injected with serial dilutions of A. baumannii WT to determine the LD 50 which was calculated to be 6.49×10 4 CFU @ 18 hrs post infection. B) Murine competition septicemia between A. baumannii WT and ΔpglL . Groups of 3 mice were injected with ∼1∶1 WT to Δ pglL CFU's and were sacrificed after 18 hrs, spleens were harvested, and bacterial load determined.

    Techniques Used: Mouse Assay, Injection, Infection

    4) Product Images from "Prophage Induction and Differential RecA and UmuDAb Transcriptome Regulation in the DNA Damage Responses of Acinetobacter baumannii and Acinetobacter baylyi"

    Article Title: Prophage Induction and Differential RecA and UmuDAb Transcriptome Regulation in the DNA Damage Responses of Acinetobacter baumannii and Acinetobacter baylyi

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0093861

    RT-qPCR experiments indicate that umuDAb is required for repression of error-prone polymerase components, not all DNA damage-induced genes. Delta Cq values from RT-qPCR experiments measuring expression of selected A. baumannii ATCC 17978 genes demonstrates the repressing activity of UmuDAb only for error prone polymerase components. The expression of each gene in both wild type and umuDAb null mutant is shown, with gene identity and A1S number listed on the x axis. Each gene was assayed in one RT-qPCR experiment (plate), with error bars indicating standard error of the mean from technical triplicates of biological triplicates.
    Figure Legend Snippet: RT-qPCR experiments indicate that umuDAb is required for repression of error-prone polymerase components, not all DNA damage-induced genes. Delta Cq values from RT-qPCR experiments measuring expression of selected A. baumannii ATCC 17978 genes demonstrates the repressing activity of UmuDAb only for error prone polymerase components. The expression of each gene in both wild type and umuDAb null mutant is shown, with gene identity and A1S number listed on the x axis. Each gene was assayed in one RT-qPCR experiment (plate), with error bars indicating standard error of the mean from technical triplicates of biological triplicates.

    Techniques Used: Quantitative RT-PCR, Expressing, Activity Assay, Mutagenesis

    Comparison of the ADP1 and 17978 chromosomal regions indicated as putative prophages. This to-scale diagram indicates the size and predicted gene functions from PHAST analysis within the chromosomal regions designated as putative prophages. Locations in the genome are represented by the ACIAD and A1S gene designation boundaries for ADP1 and 17978, respectively. ( A ) All genes represented in the ADP1 prophage region were not induced after DNA damage, and approximately one-third of the genes in this region were either hypothetical conserved phage genes, or genes of predicted phage function (with only 2 each of DNA metabolism/replication, capsid/packaging and tail genes). ( B ) Three chromosomal regions of induced genes in A. baumannii ATCC 17978 overlapped with three regions of the genome designated as putative prophages by our PHAST analysis [31] and as cryptic prophages CP5, CP9 and CP14 [38] . Genes that were not induced in this study are marked with asterisks. Previously described virulence-associated esv genes [27] are named below each gene, and error-prone polymerase alleles are shown in bright cyan color (see legend). The specific gene function of each gene in order of its placement in each prophage is described in Table 4 .
    Figure Legend Snippet: Comparison of the ADP1 and 17978 chromosomal regions indicated as putative prophages. This to-scale diagram indicates the size and predicted gene functions from PHAST analysis within the chromosomal regions designated as putative prophages. Locations in the genome are represented by the ACIAD and A1S gene designation boundaries for ADP1 and 17978, respectively. ( A ) All genes represented in the ADP1 prophage region were not induced after DNA damage, and approximately one-third of the genes in this region were either hypothetical conserved phage genes, or genes of predicted phage function (with only 2 each of DNA metabolism/replication, capsid/packaging and tail genes). ( B ) Three chromosomal regions of induced genes in A. baumannii ATCC 17978 overlapped with three regions of the genome designated as putative prophages by our PHAST analysis [31] and as cryptic prophages CP5, CP9 and CP14 [38] . Genes that were not induced in this study are marked with asterisks. Previously described virulence-associated esv genes [27] are named below each gene, and error-prone polymerase alleles are shown in bright cyan color (see legend). The specific gene function of each gene in order of its placement in each prophage is described in Table 4 .

    Techniques Used:

    5) Product Images from "Virstatin inhibits biofilm formation and motility of Acinetobacter baumannii"

    Article Title: Virstatin inhibits biofilm formation and motility of Acinetobacter baumannii

    Journal: BMC Microbiology

    doi: 10.1186/1471-2180-14-62

    Effect of virstatin on biofilms formed by A. baumannii . A - Inhibition effect on A. baumannii ATCC 17978 biofilm development in static mode: percentages measured the 24 h-biofilm formation in presence of different concentrations of virstatin (and DMSO as control) compared to biofilm formation without virstatin. Experiments were performed in triplicate and results are presented as (mean ± standard error of mean). “***” for P
    Figure Legend Snippet: Effect of virstatin on biofilms formed by A. baumannii . A - Inhibition effect on A. baumannii ATCC 17978 biofilm development in static mode: percentages measured the 24 h-biofilm formation in presence of different concentrations of virstatin (and DMSO as control) compared to biofilm formation without virstatin. Experiments were performed in triplicate and results are presented as (mean ± standard error of mean). “***” for P

    Techniques Used: Inhibition

    Virstatin affects bacterial motility and pili production. A B - Plates of motility of A. baumannii ATCC 17978 strain in presence (B) or absence (A) of virstatin in semi-solid 0.3% TSA after 24 h growth at 37°C. Typical plates are shown. The diameter of the motility zone decreased when medium is supplemented with 100 μM virstatin. The experiments were performed in triplicate. C-F - AFM images of A. baumannii ATCC 17978 water-facing side pellicles after 8 h growth: C E without virstatin; D F with 100 μM virstatin. Topographic representation using an uncommon scale made to emphasize the bacterial pili and the ‘EPS’ (extrapolymeric substances) surrounding layer.
    Figure Legend Snippet: Virstatin affects bacterial motility and pili production. A B - Plates of motility of A. baumannii ATCC 17978 strain in presence (B) or absence (A) of virstatin in semi-solid 0.3% TSA after 24 h growth at 37°C. Typical plates are shown. The diameter of the motility zone decreased when medium is supplemented with 100 μM virstatin. The experiments were performed in triplicate. C-F - AFM images of A. baumannii ATCC 17978 water-facing side pellicles after 8 h growth: C E without virstatin; D F with 100 μM virstatin. Topographic representation using an uncommon scale made to emphasize the bacterial pili and the ‘EPS’ (extrapolymeric substances) surrounding layer.

    Techniques Used:

    6) Product Images from "Impact of Acinetobacter baumannii Superoxide Dismutase on Motility, Virulence, Oxidative Stress Resistance and Susceptibility to Antibiotics"

    Article Title: Impact of Acinetobacter baumannii Superoxide Dismutase on Motility, Virulence, Oxidative Stress Resistance and Susceptibility to Antibiotics

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0101033

    A. baumannii sod2343 mutants are attenuated in the Galleria mellonella infection model. (A) Survival of Galleria caterpillars injected with 3×10 5 CFU of ATCC 17978 sod2343::Km (red), ATCC 17978 (green) and PBS (blue). (B) Infection with 1.5×10 6 CFU of 07–095 and its sod2343::Km derivative. (C) Infection with 3×10 5 CFU of 07–102 and its sod2343::Km mutant. Results represent means and standard deviations of at least three independent experiments of 16 larvae per group.
    Figure Legend Snippet: A. baumannii sod2343 mutants are attenuated in the Galleria mellonella infection model. (A) Survival of Galleria caterpillars injected with 3×10 5 CFU of ATCC 17978 sod2343::Km (red), ATCC 17978 (green) and PBS (blue). (B) Infection with 1.5×10 6 CFU of 07–095 and its sod2343::Km derivative. (C) Infection with 3×10 5 CFU of 07–102 and its sod2343::Km mutant. Results represent means and standard deviations of at least three independent experiments of 16 larvae per group.

    Techniques Used: Infection, Injection, Mutagenesis

    Recovery of A. baumannii sod2343 mutant from Galleria mellonella larvae is minor compared to parental strain. Groups of 10 larvae were injected with 1.5×10 6 CFU of ATCC 17978, ATCC 17978 sod2343::Km , and ATCC 17978 sod2343::Km pWH sod2343 , respectively. Directly after injection of the 10 larvae within a group, the larvae were frozen at −80°C, homogenized and serial dilutions were plated on CHROMagar Acinetobacter to determine CFUs. Boxes indicate the 25–75% region, the median is indicated by a line in the box, the average is indicated by a small square, whiskers indicate maximum and minimum values, respectively.
    Figure Legend Snippet: Recovery of A. baumannii sod2343 mutant from Galleria mellonella larvae is minor compared to parental strain. Groups of 10 larvae were injected with 1.5×10 6 CFU of ATCC 17978, ATCC 17978 sod2343::Km , and ATCC 17978 sod2343::Km pWH sod2343 , respectively. Directly after injection of the 10 larvae within a group, the larvae were frozen at −80°C, homogenized and serial dilutions were plated on CHROMagar Acinetobacter to determine CFUs. Boxes indicate the 25–75% region, the median is indicated by a line in the box, the average is indicated by a small square, whiskers indicate maximum and minimum values, respectively.

    Techniques Used: Mutagenesis, Injection

    7) Product Images from "Investigation of the human pathogen Acinetobacter baumannii under iron limiting conditions"

    Article Title: Investigation of the human pathogen Acinetobacter baumannii under iron limiting conditions

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-12-126

    The optimized A. baumannii FUR motif . Per position, the size of the nucleotide (T in red, A in green, C in blue and G in black) indicates its prevalence in the 22 included sequences from Table 1. The motif shows a palindrome with a central non-conserved nucleotide in position 13 which is indicated by the star. The figure of the A. baumannii ATCC 17978 FUR motif was created using WebLogo 3.0 [ 64 ].
    Figure Legend Snippet: The optimized A. baumannii FUR motif . Per position, the size of the nucleotide (T in red, A in green, C in blue and G in black) indicates its prevalence in the 22 included sequences from Table 1. The motif shows a palindrome with a central non-conserved nucleotide in position 13 which is indicated by the star. The figure of the A. baumannii ATCC 17978 FUR motif was created using WebLogo 3.0 [ 64 ].

    Techniques Used:

    Swarming of A. baumannii ATCC 17978 . A. baumannii colony material was spotted on Luria-Bertani medium containing 0.25% agar. Swarming motility (A) is visible as the channel-like growth around the dense white colony material. The absence of halo growth around the colony (B) indicates lack of swarming motility of A. baumannii when available iron is limited.
    Figure Legend Snippet: Swarming of A. baumannii ATCC 17978 . A. baumannii colony material was spotted on Luria-Bertani medium containing 0.25% agar. Swarming motility (A) is visible as the channel-like growth around the dense white colony material. The absence of halo growth around the colony (B) indicates lack of swarming motility of A. baumannii when available iron is limited.

    Techniques Used:

    Overview of transcriptional responses to iron starvation . The A. baumannii ATCC 17978 transcriptome was compared under iron replete and iron limiting conditions (200 μM DIP). All 3367 genes of the A. baumannii ATCC 17978 genome are represented on the X-axis, ordered according locus tag. Differential expression levels between iron replete and iron limiting conditions are displayed in Log 2 -values on the Y-axis. Up- and down-regulated genes under iron limiting conditions are displayed in the green and red sections, respectively. Gene names or functions have been provided for various highly differentially expressed genes, such as siderophore biosynthesis genes and the fumarases. The putative FUR binding sites displayed in the top section were identified as described in Methods and the nucleotide sequences are listed in Additional file 2 .
    Figure Legend Snippet: Overview of transcriptional responses to iron starvation . The A. baumannii ATCC 17978 transcriptome was compared under iron replete and iron limiting conditions (200 μM DIP). All 3367 genes of the A. baumannii ATCC 17978 genome are represented on the X-axis, ordered according locus tag. Differential expression levels between iron replete and iron limiting conditions are displayed in Log 2 -values on the Y-axis. Up- and down-regulated genes under iron limiting conditions are displayed in the green and red sections, respectively. Gene names or functions have been provided for various highly differentially expressed genes, such as siderophore biosynthesis genes and the fumarases. The putative FUR binding sites displayed in the top section were identified as described in Methods and the nucleotide sequences are listed in Additional file 2 .

    Techniques Used: Expressing, Binding Assay

    Transcriptional profiling of three siderophore gene clusters identified in A. baumannii ATCC 17978 . Transcriptional alteration of the three siderophore gene clusters to low iron conditions are shown, (A) siderophore gene cluster 1 (A1S_1647-1657), (B) siderophore cluster 2 (A1S_2562-2581) and (C) the acinetobactin gene cluster (A1S_2372-2392). The top arrows show predicted gene function; siderophore biosynthesis in purple, receptors and uptake mechanisms in light orange, efflux pumps in orange and genes of unknown function in white. The relative transcriptional differences between A. baumannii grown under iron replete and iron limiting conditions are depicted in the bottom set of arrows according to the green color scale bar, all values are in times-fold difference. Genes depicted in white were not differentially expressed and those in dark green were overexpressed more than 25-fold. No significant down-regulation was observed within the siderophore gene clusters. Putative FUR boxes are shown as black arrows.
    Figure Legend Snippet: Transcriptional profiling of three siderophore gene clusters identified in A. baumannii ATCC 17978 . Transcriptional alteration of the three siderophore gene clusters to low iron conditions are shown, (A) siderophore gene cluster 1 (A1S_1647-1657), (B) siderophore cluster 2 (A1S_2562-2581) and (C) the acinetobactin gene cluster (A1S_2372-2392). The top arrows show predicted gene function; siderophore biosynthesis in purple, receptors and uptake mechanisms in light orange, efflux pumps in orange and genes of unknown function in white. The relative transcriptional differences between A. baumannii grown under iron replete and iron limiting conditions are depicted in the bottom set of arrows according to the green color scale bar, all values are in times-fold difference. Genes depicted in white were not differentially expressed and those in dark green were overexpressed more than 25-fold. No significant down-regulation was observed within the siderophore gene clusters. Putative FUR boxes are shown as black arrows.

    Techniques Used:

    8) Product Images from "Transcriptome profiling in imipenem-selected Acinetobacter baumannii"

    Article Title: Transcriptome profiling in imipenem-selected Acinetobacter baumannii

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-15-815

    LC-MS/MS chromatogram of imipenem under co-incubating with A. baumannii ATCC 17978, IPM-2 m and IPM-8 m cells. Control, imipenem standard solution.
    Figure Legend Snippet: LC-MS/MS chromatogram of imipenem under co-incubating with A. baumannii ATCC 17978, IPM-2 m and IPM-8 m cells. Control, imipenem standard solution.

    Techniques Used: Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry

    Validation of the transcriptome results. The transcriptomic results obtained through RNA sequencing were validated using qualitative RT-PCR (RT-qPCR) analysis. The level of differential expression of eight genes was compared, showing a correlation between RNA sequencing (Y-axis) and RT-qPCR analysis (X-axis). The level of differential expression between A. baumannii ATCC 17978 and their mutants is given as Log 2 -values. R 2 , the coefficient of determination.
    Figure Legend Snippet: Validation of the transcriptome results. The transcriptomic results obtained through RNA sequencing were validated using qualitative RT-PCR (RT-qPCR) analysis. The level of differential expression of eight genes was compared, showing a correlation between RNA sequencing (Y-axis) and RT-qPCR analysis (X-axis). The level of differential expression between A. baumannii ATCC 17978 and their mutants is given as Log 2 -values. R 2 , the coefficient of determination.

    Techniques Used: RNA Sequencing Assay, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR, Expressing

    9) Product Images from "Genetic Dissection of the Type VI Secretion System in Acinetobacter and Identification of a Novel Peptidoglycan Hydrolase, TagX, Required for Its Biogenesis"

    Article Title: Genetic Dissection of the Type VI Secretion System in Acinetobacter and Identification of a Novel Peptidoglycan Hydrolase, TagX, Required for Its Biogenesis

    Journal: mBio

    doi: 10.1128/mBio.01253-16

    Hcp secretion and bacterial killing by T6 + A. baumannii ATCC 17978 vgrG mutants. Western blot assays probing for Hcp and RNA polymerase (RNAP) and E. coli killing assays were performed with single vgrG deletion mutants (A), double vgrG deletion mutants (B), triple and quadruple vgrG deletion mutants (C), and tse2 and tse3 effector mutants (D) in the Δ vgrG34 or Δ vgrG24 background, respectively. For the bacterial killing assay graphs, the A. baumannii predator strains indicated are on the x axis and the log-transformed surviving E. coli MG1655R CFU count is on the y axis. A drug-sensitive strain of E. coli or no predator (none) was used as a control. The results shown are from two independent experiments performed in triplicate. Asterisks indicate statistical significance compared to wild-type T6 + (unpaired, two-tailed Student t test; **, P
    Figure Legend Snippet: Hcp secretion and bacterial killing by T6 + A. baumannii ATCC 17978 vgrG mutants. Western blot assays probing for Hcp and RNA polymerase (RNAP) and E. coli killing assays were performed with single vgrG deletion mutants (A), double vgrG deletion mutants (B), triple and quadruple vgrG deletion mutants (C), and tse2 and tse3 effector mutants (D) in the Δ vgrG34 or Δ vgrG24 background, respectively. For the bacterial killing assay graphs, the A. baumannii predator strains indicated are on the x axis and the log-transformed surviving E. coli MG1655R CFU count is on the y axis. A drug-sensitive strain of E. coli or no predator (none) was used as a control. The results shown are from two independent experiments performed in triplicate. Asterisks indicate statistical significance compared to wild-type T6 + (unpaired, two-tailed Student t test; **, P

    Techniques Used: Western Blot, Transformation Assay, Two Tailed Test

    The TagX protein of A. baumannii ATCC 17978 . (A) At the top is a schematic of TagX showing the locations of the predicted transmembrane (TM) domains and the pfam13539 peptidase domain, which resembles members of the VanY superfamily. At the bottom is an alignment of the A. baumannii and A. baylyi TagX pfam13539 domains with the Listeria phage endolysin PlyA500 (accession no. AAY52812 ). Relevant metal-coordinating residues are indicated by the blue arrows, and the red arrow shows the general base in the active site of the characterized Ply500 enzyme. (B, i) The crystal structure of the enzymatically active domain Ply500 (Protein Data Bank code 2VO9 ) ( 53 , 54 ) (left) and the modeled structure of the C-terminal end of A. baumannii TagX. The metal-coordinating residues are red, and the general base residue is purple. (B, ii) Overlay of the crystal structure of Ply500 and the modeled structure of TagX. Modeling was performed with SWISS-MODEL ( 80 ).
    Figure Legend Snippet: The TagX protein of A. baumannii ATCC 17978 . (A) At the top is a schematic of TagX showing the locations of the predicted transmembrane (TM) domains and the pfam13539 peptidase domain, which resembles members of the VanY superfamily. At the bottom is an alignment of the A. baumannii and A. baylyi TagX pfam13539 domains with the Listeria phage endolysin PlyA500 (accession no. AAY52812 ). Relevant metal-coordinating residues are indicated by the blue arrows, and the red arrow shows the general base in the active site of the characterized Ply500 enzyme. (B, i) The crystal structure of the enzymatically active domain Ply500 (Protein Data Bank code 2VO9 ) ( 53 , 54 ) (left) and the modeled structure of the C-terminal end of A. baumannii TagX. The metal-coordinating residues are red, and the general base residue is purple. (B, ii) Overlay of the crystal structure of Ply500 and the modeled structure of TagX. Modeling was performed with SWISS-MODEL ( 80 ).

    Techniques Used:

    (A) Schematic of the core T6SS clusters from A. baumannii ATCC 17978, B. thailandensis E264, and R. solanacearum GMI1000. Black arrows represent T6SS-related genes, and tagX is blue. Genes with unknown functions are gray. (B) Alignment of the full-length TagX proteins from these three bacteria with the predicted transmembrane residues of the A. baumannii TagX underlined.
    Figure Legend Snippet: (A) Schematic of the core T6SS clusters from A. baumannii ATCC 17978, B. thailandensis E264, and R. solanacearum GMI1000. Black arrows represent T6SS-related genes, and tagX is blue. Genes with unknown functions are gray. (B) Alignment of the full-length TagX proteins from these three bacteria with the predicted transmembrane residues of the A. baumannii TagX underlined.

    Techniques Used:

    10) Product Images from "Genetic Dissection of the Type VI Secretion System in Acinetobacter and Identification of a Novel Peptidoglycan Hydrolase, TagX, Required for Its Biogenesis"

    Article Title: Genetic Dissection of the Type VI Secretion System in Acinetobacter and Identification of a Novel Peptidoglycan Hydrolase, TagX, Required for Its Biogenesis

    Journal: mBio

    doi: 10.1128/mBio.01253-16

    Hcp secretion and bacterial killing by T6 + A. baumannii ATCC 17978 vgrG mutants. Western blot assays probing for Hcp and RNA polymerase (RNAP) and E. coli killing assays were performed with single vgrG deletion mutants (A), double vgrG deletion mutants (B), triple and quadruple vgrG deletion mutants (C), and tse2 and tse3 effector mutants (D) in the Δ vgrG34 or Δ vgrG24 background, respectively. For the bacterial killing assay graphs, the A. baumannii predator strains indicated are on the x axis and the log-transformed surviving E. coli MG1655R CFU count is on the y axis. A drug-sensitive strain of E. coli or no predator (none) was used as a control. The results shown are from two independent experiments performed in triplicate. Asterisks indicate statistical significance compared to wild-type T6 + (unpaired, two-tailed Student t test; **, P
    Figure Legend Snippet: Hcp secretion and bacterial killing by T6 + A. baumannii ATCC 17978 vgrG mutants. Western blot assays probing for Hcp and RNA polymerase (RNAP) and E. coli killing assays were performed with single vgrG deletion mutants (A), double vgrG deletion mutants (B), triple and quadruple vgrG deletion mutants (C), and tse2 and tse3 effector mutants (D) in the Δ vgrG34 or Δ vgrG24 background, respectively. For the bacterial killing assay graphs, the A. baumannii predator strains indicated are on the x axis and the log-transformed surviving E. coli MG1655R CFU count is on the y axis. A drug-sensitive strain of E. coli or no predator (none) was used as a control. The results shown are from two independent experiments performed in triplicate. Asterisks indicate statistical significance compared to wild-type T6 + (unpaired, two-tailed Student t test; **, P

    Techniques Used: Western Blot, Transformation Assay, Two Tailed Test

    The TagX protein of A. baumannii ATCC 17978 . (A) At the top is a schematic of TagX showing the locations of the predicted transmembrane (TM) domains and the pfam13539 peptidase domain, which resembles members of the VanY superfamily. At the bottom is an alignment of the A. baumannii and A. baylyi TagX pfam13539 domains with the Listeria phage endolysin PlyA500 (accession no. AAY52812 ). Relevant metal-coordinating residues are indicated by the blue arrows, and the red arrow shows the general base in the active site of the characterized Ply500 enzyme. (B, i) The crystal structure of the enzymatically active domain Ply500 (Protein Data Bank code 2VO9 ) ( 53 , 54 ) (left) and the modeled structure of the C-terminal end of A. baumannii TagX. The metal-coordinating residues are red, and the general base residue is purple. (B, ii) Overlay of the crystal structure of Ply500 and the modeled structure of TagX. Modeling was performed with SWISS-MODEL ( 80 ).
    Figure Legend Snippet: The TagX protein of A. baumannii ATCC 17978 . (A) At the top is a schematic of TagX showing the locations of the predicted transmembrane (TM) domains and the pfam13539 peptidase domain, which resembles members of the VanY superfamily. At the bottom is an alignment of the A. baumannii and A. baylyi TagX pfam13539 domains with the Listeria phage endolysin PlyA500 (accession no. AAY52812 ). Relevant metal-coordinating residues are indicated by the blue arrows, and the red arrow shows the general base in the active site of the characterized Ply500 enzyme. (B, i) The crystal structure of the enzymatically active domain Ply500 (Protein Data Bank code 2VO9 ) ( 53 , 54 ) (left) and the modeled structure of the C-terminal end of A. baumannii TagX. The metal-coordinating residues are red, and the general base residue is purple. (B, ii) Overlay of the crystal structure of Ply500 and the modeled structure of TagX. Modeling was performed with SWISS-MODEL ( 80 ).

    Techniques Used:

    (A) Schematic of the core T6SS clusters from A. baumannii ATCC 17978, B. thailandensis E264, and R. solanacearum GMI1000. Black arrows represent T6SS-related genes, and tagX is blue. Genes with unknown functions are gray. (B) Alignment of the full-length TagX proteins from these three bacteria with the predicted transmembrane residues of the A. baumannii TagX underlined.
    Figure Legend Snippet: (A) Schematic of the core T6SS clusters from A. baumannii ATCC 17978, B. thailandensis E264, and R. solanacearum GMI1000. Black arrows represent T6SS-related genes, and tagX is blue. Genes with unknown functions are gray. (B) Alignment of the full-length TagX proteins from these three bacteria with the predicted transmembrane residues of the A. baumannii TagX underlined.

    Techniques Used:

    11) Product Images from "Zinc stress induces copper depletion in Acinetobacter baumannii"

    Article Title: Zinc stress induces copper depletion in Acinetobacter baumannii

    Journal: BMC Microbiology

    doi: 10.1186/s12866-017-0965-y

    The effect of Zn and Cu treatment on the resistance of A. baumannii to oxidative stress. The effect of ( a ) 40 μM paraquat and ( e ) 160 μM H 2 O 2 on A. baumannii strain ATCC 17978 grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions, was determined by measuring the optical density at 600 nm (OD 600 ). The growth delay between untreated (UT) and paraquat- or H 2 O 2 -treated cells was examined by comparing the EC 50 under each relevant metal ion stress condition. Statistical analyses were performed using a one-way ANOVA. b The total SOD activity of mid-log phase cells (untreated, 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu) was determined. Data were corrected for total protein content, followed by normalisation to untreated cells. Statistical analyses were performed using a Student’s t -test. The mRNA transcription levels of ( c ) Cu-Zn superoxide dismutase (Cu-Zn-SOD; A1S_3143), ( d ) Fe/Mn superoxide dismutase (Fe/Mn-SOD; A1S_2343) were determined by qRT-PCR. The transcription levels examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu were corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest. For all statistical analyses; ns = not significant, * = p
    Figure Legend Snippet: The effect of Zn and Cu treatment on the resistance of A. baumannii to oxidative stress. The effect of ( a ) 40 μM paraquat and ( e ) 160 μM H 2 O 2 on A. baumannii strain ATCC 17978 grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions, was determined by measuring the optical density at 600 nm (OD 600 ). The growth delay between untreated (UT) and paraquat- or H 2 O 2 -treated cells was examined by comparing the EC 50 under each relevant metal ion stress condition. Statistical analyses were performed using a one-way ANOVA. b The total SOD activity of mid-log phase cells (untreated, 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu) was determined. Data were corrected for total protein content, followed by normalisation to untreated cells. Statistical analyses were performed using a Student’s t -test. The mRNA transcription levels of ( c ) Cu-Zn superoxide dismutase (Cu-Zn-SOD; A1S_3143), ( d ) Fe/Mn superoxide dismutase (Fe/Mn-SOD; A1S_2343) were determined by qRT-PCR. The transcription levels examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu were corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest. For all statistical analyses; ns = not significant, * = p

    Techniques Used: Activity Assay, Quantitative RT-PCR

    An overview of the cellular responses of A. baumannii to Zn and Cu stress. Cartoon representation of A. baumannii ATCC 17978 under either high Zn (400 μM), high Cu (400 μM) or high Zn + Cu (400 μM each) conditions based on the observations from this work. The numbers within the efflux systems (ovals) are the locus-tags for A. baumannii strain ATCC 17978, excluding their prefix “A1S_”. The efflux proteins in red are those up-regulated under high Zn, in blue by high Cu and those in purple were found to be transcriptionally responsive to both high Zn and Cu independently (A1S_2939 and A1S_0709) or the combination of high Zn and high Cu (A1S_2939 and A1S_3098). Under high Zn and high Zn + Cu conditions, A. baumannii becomes increasingly sensitive to superoxide stress (O 2 .- in the red burst), but shows increased resistance to hydrogen peroxide (green cloud). Under high Cu conditions, A. baumannii shows increased susceptibility to hydrogen peroxide (H 2 O 2 in the red burst). Under high Zn conditions and high Zn + Cu conditions, Cu levels reduce to 74 and 62%, respectively
    Figure Legend Snippet: An overview of the cellular responses of A. baumannii to Zn and Cu stress. Cartoon representation of A. baumannii ATCC 17978 under either high Zn (400 μM), high Cu (400 μM) or high Zn + Cu (400 μM each) conditions based on the observations from this work. The numbers within the efflux systems (ovals) are the locus-tags for A. baumannii strain ATCC 17978, excluding their prefix “A1S_”. The efflux proteins in red are those up-regulated under high Zn, in blue by high Cu and those in purple were found to be transcriptionally responsive to both high Zn and Cu independently (A1S_2939 and A1S_0709) or the combination of high Zn and high Cu (A1S_2939 and A1S_3098). Under high Zn and high Zn + Cu conditions, A. baumannii becomes increasingly sensitive to superoxide stress (O 2 .- in the red burst), but shows increased resistance to hydrogen peroxide (green cloud). Under high Cu conditions, A. baumannii shows increased susceptibility to hydrogen peroxide (H 2 O 2 in the red burst). Under high Zn conditions and high Zn + Cu conditions, Cu levels reduce to 74 and 62%, respectively

    Techniques Used:

    The effects of Zn and Cu on A. baumannii membrane biology. The major cellular fatty acid constituents of A. baumannii strain ATCC 17978, grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions were determined by gas chromatography. The abundance of fatty acids, expressed as percentage of total cellular fatty acids, is ( a ) 16:0 ( b ) 18:0, ( c ) 16:1n-7 and ( d ) 18:1n-7. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, ** = p
    Figure Legend Snippet: The effects of Zn and Cu on A. baumannii membrane biology. The major cellular fatty acid constituents of A. baumannii strain ATCC 17978, grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions were determined by gas chromatography. The abundance of fatty acids, expressed as percentage of total cellular fatty acids, is ( a ) 16:0 ( b ) 18:0, ( c ) 16:1n-7 and ( d ) 18:1n-7. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, ** = p

    Techniques Used: Gas Chromatography

    The effect of Zn and Cu on macrophage-mediated killing of A. baumannii . The survival of A. baumannii ATCC 17978 cells in the presence of THP-1 human monocyte-derived macrophages that were either treated with 50 μM Zn, 50 μM Cu or 50 μM Zn + 50 μM Cu was compared to survival of A. baumannii ATCC 17978 cells in the presence of untreated THP-1 cells. The data are the mean of biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant, * = p
    Figure Legend Snippet: The effect of Zn and Cu on macrophage-mediated killing of A. baumannii . The survival of A. baumannii ATCC 17978 cells in the presence of THP-1 human monocyte-derived macrophages that were either treated with 50 μM Zn, 50 μM Cu or 50 μM Zn + 50 μM Cu was compared to survival of A. baumannii ATCC 17978 cells in the presence of untreated THP-1 cells. The data are the mean of biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant, * = p

    Techniques Used: Derivative Assay, Two Tailed Test

    The effect of Zn and Cu stress on A. baumannii growth and metal ion homeostasis. Growth as determined by measuring the optical density at 600 nm (OD 600 ) of A. baumannii strain ATCC 17978 under increasing concentrations of ( a ) Zn, ( b ) Cu, or ( c ) Zn + Cu ( n ≥ 3). Examination of the ( d ) Zn, ( e ) Cu and ( f ) Fe levels in untreated cells and cells grown in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu. The cellular metal ion content (in microgram) per gram of desiccated cells was determined by ICP-MS. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant and * = p
    Figure Legend Snippet: The effect of Zn and Cu stress on A. baumannii growth and metal ion homeostasis. Growth as determined by measuring the optical density at 600 nm (OD 600 ) of A. baumannii strain ATCC 17978 under increasing concentrations of ( a ) Zn, ( b ) Cu, or ( c ) Zn + Cu ( n ≥ 3). Examination of the ( d ) Zn, ( e ) Cu and ( f ) Fe levels in untreated cells and cells grown in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu. The cellular metal ion content (in microgram) per gram of desiccated cells was determined by ICP-MS. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant and * = p

    Techniques Used: Mass Spectrometry, Two Tailed Test

    Transcriptional analyses of putative A. baumannii Zn and Cu efflux systems. The mRNA transcription levels of the putative ( a ) Cation Diffusion Facilitator (CDF), ( b ) Heavy Metal Efflux (HME), ( c ) P-type ATPase, ( d ) CorA-type and ( e ) CopB-like copper resistance genes of A. baumannii strain ATCC 17978 were determined by qRT-PCR. Transcription levels were examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu and corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, * = p
    Figure Legend Snippet: Transcriptional analyses of putative A. baumannii Zn and Cu efflux systems. The mRNA transcription levels of the putative ( a ) Cation Diffusion Facilitator (CDF), ( b ) Heavy Metal Efflux (HME), ( c ) P-type ATPase, ( d ) CorA-type and ( e ) CopB-like copper resistance genes of A. baumannii strain ATCC 17978 were determined by qRT-PCR. Transcription levels were examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu and corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, * = p

    Techniques Used: Diffusion-based Assay, Quantitative RT-PCR

    12) Product Images from "Zinc stress induces copper depletion in Acinetobacter baumannii"

    Article Title: Zinc stress induces copper depletion in Acinetobacter baumannii

    Journal: BMC Microbiology

    doi: 10.1186/s12866-017-0965-y

    The effect of Zn and Cu treatment on the resistance of A. baumannii to oxidative stress. The effect of ( a ) 40 μM paraquat and ( e ) 160 μM H 2 O 2 on A. baumannii strain ATCC 17978 grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions, was determined by measuring the optical density at 600 nm (OD 600 ). The growth delay between untreated (UT) and paraquat- or H 2 O 2 -treated cells was examined by comparing the EC 50 under each relevant metal ion stress condition. Statistical analyses were performed using a one-way ANOVA. b The total SOD activity of mid-log phase cells (untreated, 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu) was determined. Data were corrected for total protein content, followed by normalisation to untreated cells. Statistical analyses were performed using a Student’s t -test. The mRNA transcription levels of ( c ) Cu-Zn superoxide dismutase (Cu-Zn-SOD; A1S_3143), ( d ) Fe/Mn superoxide dismutase (Fe/Mn-SOD; A1S_2343) were determined by qRT-PCR. The transcription levels examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu were corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest. For all statistical analyses; ns = not significant, * = p
    Figure Legend Snippet: The effect of Zn and Cu treatment on the resistance of A. baumannii to oxidative stress. The effect of ( a ) 40 μM paraquat and ( e ) 160 μM H 2 O 2 on A. baumannii strain ATCC 17978 grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions, was determined by measuring the optical density at 600 nm (OD 600 ). The growth delay between untreated (UT) and paraquat- or H 2 O 2 -treated cells was examined by comparing the EC 50 under each relevant metal ion stress condition. Statistical analyses were performed using a one-way ANOVA. b The total SOD activity of mid-log phase cells (untreated, 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu) was determined. Data were corrected for total protein content, followed by normalisation to untreated cells. Statistical analyses were performed using a Student’s t -test. The mRNA transcription levels of ( c ) Cu-Zn superoxide dismutase (Cu-Zn-SOD; A1S_3143), ( d ) Fe/Mn superoxide dismutase (Fe/Mn-SOD; A1S_2343) were determined by qRT-PCR. The transcription levels examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu were corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest. For all statistical analyses; ns = not significant, * = p

    Techniques Used: Activity Assay, Quantitative RT-PCR

    An overview of the cellular responses of A. baumannii to Zn and Cu stress. Cartoon representation of A. baumannii ATCC 17978 under either high Zn (400 μM), high Cu (400 μM) or high Zn + Cu (400 μM each) conditions based on the observations from this work. The numbers within the efflux systems (ovals) are the locus-tags for A. baumannii strain ATCC 17978, excluding their prefix “A1S_”. The efflux proteins in red are those up-regulated under high Zn, in blue by high Cu and those in purple were found to be transcriptionally responsive to both high Zn and Cu independently (A1S_2939 and A1S_0709) or the combination of high Zn and high Cu (A1S_2939 and A1S_3098). Under high Zn and high Zn + Cu conditions, A. baumannii becomes increasingly sensitive to superoxide stress (O 2 .- in the red burst), but shows increased resistance to hydrogen peroxide (green cloud). Under high Cu conditions, A. baumannii shows increased susceptibility to hydrogen peroxide (H 2 O 2 in the red burst). Under high Zn conditions and high Zn + Cu conditions, Cu levels reduce to 74 and 62%, respectively
    Figure Legend Snippet: An overview of the cellular responses of A. baumannii to Zn and Cu stress. Cartoon representation of A. baumannii ATCC 17978 under either high Zn (400 μM), high Cu (400 μM) or high Zn + Cu (400 μM each) conditions based on the observations from this work. The numbers within the efflux systems (ovals) are the locus-tags for A. baumannii strain ATCC 17978, excluding their prefix “A1S_”. The efflux proteins in red are those up-regulated under high Zn, in blue by high Cu and those in purple were found to be transcriptionally responsive to both high Zn and Cu independently (A1S_2939 and A1S_0709) or the combination of high Zn and high Cu (A1S_2939 and A1S_3098). Under high Zn and high Zn + Cu conditions, A. baumannii becomes increasingly sensitive to superoxide stress (O 2 .- in the red burst), but shows increased resistance to hydrogen peroxide (green cloud). Under high Cu conditions, A. baumannii shows increased susceptibility to hydrogen peroxide (H 2 O 2 in the red burst). Under high Zn conditions and high Zn + Cu conditions, Cu levels reduce to 74 and 62%, respectively

    Techniques Used:

    The effects of Zn and Cu on A. baumannii membrane biology. The major cellular fatty acid constituents of A. baumannii strain ATCC 17978, grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions were determined by gas chromatography. The abundance of fatty acids, expressed as percentage of total cellular fatty acids, is ( a ) 16:0 ( b ) 18:0, ( c ) 16:1n-7 and ( d ) 18:1n-7. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, ** = p
    Figure Legend Snippet: The effects of Zn and Cu on A. baumannii membrane biology. The major cellular fatty acid constituents of A. baumannii strain ATCC 17978, grown in the presence of 400 μM Zn, 400 μM Cu, 400 μM Zn + 400 μM Cu, or without added metal ions were determined by gas chromatography. The abundance of fatty acids, expressed as percentage of total cellular fatty acids, is ( a ) 16:0 ( b ) 18:0, ( c ) 16:1n-7 and ( d ) 18:1n-7. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, ** = p

    Techniques Used: Gas Chromatography

    The effect of Zn and Cu on macrophage-mediated killing of A. baumannii . The survival of A. baumannii ATCC 17978 cells in the presence of THP-1 human monocyte-derived macrophages that were either treated with 50 μM Zn, 50 μM Cu or 50 μM Zn + 50 μM Cu was compared to survival of A. baumannii ATCC 17978 cells in the presence of untreated THP-1 cells. The data are the mean of biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant, * = p
    Figure Legend Snippet: The effect of Zn and Cu on macrophage-mediated killing of A. baumannii . The survival of A. baumannii ATCC 17978 cells in the presence of THP-1 human monocyte-derived macrophages that were either treated with 50 μM Zn, 50 μM Cu or 50 μM Zn + 50 μM Cu was compared to survival of A. baumannii ATCC 17978 cells in the presence of untreated THP-1 cells. The data are the mean of biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant, * = p

    Techniques Used: Derivative Assay, Two Tailed Test

    The effect of Zn and Cu stress on A. baumannii growth and metal ion homeostasis. Growth as determined by measuring the optical density at 600 nm (OD 600 ) of A. baumannii strain ATCC 17978 under increasing concentrations of ( a ) Zn, ( b ) Cu, or ( c ) Zn + Cu ( n ≥ 3). Examination of the ( d ) Zn, ( e ) Cu and ( f ) Fe levels in untreated cells and cells grown in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu. The cellular metal ion content (in microgram) per gram of desiccated cells was determined by ICP-MS. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant and * = p
    Figure Legend Snippet: The effect of Zn and Cu stress on A. baumannii growth and metal ion homeostasis. Growth as determined by measuring the optical density at 600 nm (OD 600 ) of A. baumannii strain ATCC 17978 under increasing concentrations of ( a ) Zn, ( b ) Cu, or ( c ) Zn + Cu ( n ≥ 3). Examination of the ( d ) Zn, ( e ) Cu and ( f ) Fe levels in untreated cells and cells grown in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu. The cellular metal ion content (in microgram) per gram of desiccated cells was determined by ICP-MS. The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed using a two-tailed Student’s t -test; ns = not significant and * = p

    Techniques Used: Mass Spectrometry, Two Tailed Test

    Transcriptional analyses of putative A. baumannii Zn and Cu efflux systems. The mRNA transcription levels of the putative ( a ) Cation Diffusion Facilitator (CDF), ( b ) Heavy Metal Efflux (HME), ( c ) P-type ATPase, ( d ) CorA-type and ( e ) CopB-like copper resistance genes of A. baumannii strain ATCC 17978 were determined by qRT-PCR. Transcription levels were examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu and corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, * = p
    Figure Legend Snippet: Transcriptional analyses of putative A. baumannii Zn and Cu efflux systems. The mRNA transcription levels of the putative ( a ) Cation Diffusion Facilitator (CDF), ( b ) Heavy Metal Efflux (HME), ( c ) P-type ATPase, ( d ) CorA-type and ( e ) CopB-like copper resistance genes of A. baumannii strain ATCC 17978 were determined by qRT-PCR. Transcription levels were examined in the presence of 400 μM Zn, 400 μM Cu or 400 μM Zn + 400 μM Cu and corrected to untreated cells following internal normalisation to GAPDH . The data are the mean of at least biological triplicates (± SEM). Statistical analyses were performed by one-way ANOVA using Dunnett’s posttest; ns = not significant, * = p

    Techniques Used: Diffusion-based Assay, Quantitative RT-PCR

    13) Product Images from "Response to Bile Salts in Clinical Strains of Acinetobacter baumannii Lacking the AdeABC Efflux Pump: Virulence Associated with Quorum Sensing"

    Article Title: Response to Bile Salts in Clinical Strains of Acinetobacter baumannii Lacking the AdeABC Efflux Pump: Virulence Associated with Quorum Sensing

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2017.00143

    SEM analysis of A. baumannii cells cultured in the absence (A,C,E,G) and presence of 0.5% bile salts (B,D,F,H) . (A,B) A. baumannii ATCC 17978; (C,D) A. baumannii Δ adeB ATCC 17978; (E,F) A. baumannii Δ adeL ATCC 17978; (G,H) Ab421 GEIH-2010. (Scale bars: 20 μm). It is observed in presence of bile salts, the state of adhesion in A. baumannii ATCC 17978 (B) , slime layer-micro colonies (previous state of biofilm formation) in A. baumannii Δ ade B ATCC 17978 (D) , proliferation in A. baumannii Δ ade L ATCC 17978 (F) and finally, biofilm formation in Ab421 GEIH-2010 (H) . The arrows indicate the most advanced stages of biofilm development.
    Figure Legend Snippet: SEM analysis of A. baumannii cells cultured in the absence (A,C,E,G) and presence of 0.5% bile salts (B,D,F,H) . (A,B) A. baumannii ATCC 17978; (C,D) A. baumannii Δ adeB ATCC 17978; (E,F) A. baumannii Δ adeL ATCC 17978; (G,H) Ab421 GEIH-2010. (Scale bars: 20 μm). It is observed in presence of bile salts, the state of adhesion in A. baumannii ATCC 17978 (B) , slime layer-micro colonies (previous state of biofilm formation) in A. baumannii Δ ade B ATCC 17978 (D) , proliferation in A. baumannii Δ ade L ATCC 17978 (F) and finally, biofilm formation in Ab421 GEIH-2010 (H) . The arrows indicate the most advanced stages of biofilm development.

    Techniques Used: Cell Culture

    Motility of A. baumannii strains in the presence of 0.5 and 1% of bile salts in three types of LB broth (Normal LB medium, Modified LB-LS and Modified LB-LN) . (a) A. baumannii ATCC 17978; (b) A. baumannii Δ adeB ATCC 17978; (c) A. baumannii Δ adeL ATCC 17978; and (d) Ab421 GEIH-2010. The arrows on the left show the similarity in the behavior of both strains ( A. baumannii Δ adeB ATCC 17978 and Ab421 GEIH-2010) in different medium and in presence of bile salts.
    Figure Legend Snippet: Motility of A. baumannii strains in the presence of 0.5 and 1% of bile salts in three types of LB broth (Normal LB medium, Modified LB-LS and Modified LB-LN) . (a) A. baumannii ATCC 17978; (b) A. baumannii Δ adeB ATCC 17978; (c) A. baumannii Δ adeL ATCC 17978; and (d) Ab421 GEIH-2010. The arrows on the left show the similarity in the behavior of both strains ( A. baumannii Δ adeB ATCC 17978 and Ab421 GEIH-2010) in different medium and in presence of bile salts.

    Techniques Used: Modification

    Quantification of biofilm formation by crystal violet staining . Eight independent replicates were considered. Results were analyzed by a Student's t -test. The values are means and bars indicate the standard deviation. Arrows indicate the highest biofilm producing strains cultured in the presence of bile salts (BS): A. baumannii Δ adeB ATCC 17978 and Ab421 GEIH-2010.
    Figure Legend Snippet: Quantification of biofilm formation by crystal violet staining . Eight independent replicates were considered. Results were analyzed by a Student's t -test. The values are means and bars indicate the standard deviation. Arrows indicate the highest biofilm producing strains cultured in the presence of bile salts (BS): A. baumannii Δ adeB ATCC 17978 and Ab421 GEIH-2010.

    Techniques Used: Staining, Standard Deviation, Cell Culture

    Curves showing growth of the A. baumannii isolates belonging to clone ST79/PFGE-HUI-1 in the presence of bile salts (0.5%) in LB-LN . A. baumannii ATCC 17978 was included as a control.
    Figure Legend Snippet: Curves showing growth of the A. baumannii isolates belonging to clone ST79/PFGE-HUI-1 in the presence of bile salts (0.5%) in LB-LN . A. baumannii ATCC 17978 was included as a control.

    Techniques Used:

    14) Product Images from "A Transcriptional Regulatory Mechanism Finely Tunes the Firing of Type VI Secretion System in Response to Bacterial Enemies"

    Article Title: A Transcriptional Regulatory Mechanism Finely Tunes the Firing of Type VI Secretion System in Response to Bacterial Enemies

    Journal: mBio

    doi: 10.1128/mBio.00559-17

    Acinetobacter killing capacity and RcsB-dependent induction of Serratia T6SS expression. (A, C) Recovery of viable S. marcescens RM66262 wt or mutant strains cocultured with wt or mutant strains of A. baumannii ATCC 17978 (A) or A. nosocomialis M2 (C) for 4 h at 37°C, with an initial 5:1 (attacker/target) ratio. E. coli DH5α mixed with target bacteria at a 5:1 ratio was used for controls. A. baumannii wt or vgrG4 strains were 200-fold more proficient in outcompeting Serratia than the other vgrG single, double, or triple mutant strains analyzed. A. nosocomialis wt was ∼5 orders of magnitude more proficient in outcompeting Serratia than the mutant strains analyzed. Average values ± SEM from four independent experiments are shown (**, P
    Figure Legend Snippet: Acinetobacter killing capacity and RcsB-dependent induction of Serratia T6SS expression. (A, C) Recovery of viable S. marcescens RM66262 wt or mutant strains cocultured with wt or mutant strains of A. baumannii ATCC 17978 (A) or A. nosocomialis M2 (C) for 4 h at 37°C, with an initial 5:1 (attacker/target) ratio. E. coli DH5α mixed with target bacteria at a 5:1 ratio was used for controls. A. baumannii wt or vgrG4 strains were 200-fold more proficient in outcompeting Serratia than the other vgrG single, double, or triple mutant strains analyzed. A. nosocomialis wt was ∼5 orders of magnitude more proficient in outcompeting Serratia than the mutant strains analyzed. Average values ± SEM from four independent experiments are shown (**, P

    Techniques Used: Expressing, Mutagenesis

    15) Product Images from "Synergy between Active Efflux and Outer Membrane Diffusion Defines Rules of Antibiotic Permeation into Gram-Negative Bacteria"

    Article Title: Synergy between Active Efflux and Outer Membrane Diffusion Defines Rules of Antibiotic Permeation into Gram-Negative Bacteria

    Journal: mBio

    doi: 10.1128/mBio.01172-17

    Intracellular uptake of the lipophilic probe NPN in A. baumannii ATCC 17978 and B. thailandensis and their hyperporinated and efflux-deficient derivatives. Data represent real-time kinetics of NPN (6 μM final concentration) uptake in Bt cells (A), P. aeruginosa PAO1 (B), A. baumannii ATCC 17978 (C), and B. cepacia ATCC 25416 (D). In all panels, blue represents parent strains, orange represents hyperporinated parent strains, gray represents efflux-deficient mutants, and yellow represents efflux-deficient hyperporinated strains.
    Figure Legend Snippet: Intracellular uptake of the lipophilic probe NPN in A. baumannii ATCC 17978 and B. thailandensis and their hyperporinated and efflux-deficient derivatives. Data represent real-time kinetics of NPN (6 μM final concentration) uptake in Bt cells (A), P. aeruginosa PAO1 (B), A. baumannii ATCC 17978 (C), and B. cepacia ATCC 25416 (D). In all panels, blue represents parent strains, orange represents hyperporinated parent strains, gray represents efflux-deficient mutants, and yellow represents efflux-deficient hyperporinated strains.

    Techniques Used: Concentration Assay

    Statistical analyses of relationships between efflux and OM ratios in different species. (A) Hierarchical clusters and the heat map of all measured efflux and OM ratios. Data represent efflux ratios (wild-type strain [WT]/efflux mutant) with native membranes and OM ratios (parent-parent-Pore) for efflux-proficient (Bt, A. baumannii ATCC 17978, PAO1, and E. coli ) and efflux-deficient (AbΔ3, PΔ3, ΔTolC, and BtΔ2) cells. The heat scale is the base-2 logarithm of the change in MICs. The identified clusters of antibiotics are numbered. (B) Principal-component analysis of antibiotic clusters with best-fit Gaussian distributions. Clusters are numbered as described for panel A. (C) Sizes [log(MW)] and hydrophobicities (logD) of antibiotics belonging to the indicated clusters and fitted to the Gaussian distributions as indicated.
    Figure Legend Snippet: Statistical analyses of relationships between efflux and OM ratios in different species. (A) Hierarchical clusters and the heat map of all measured efflux and OM ratios. Data represent efflux ratios (wild-type strain [WT]/efflux mutant) with native membranes and OM ratios (parent-parent-Pore) for efflux-proficient (Bt, A. baumannii ATCC 17978, PAO1, and E. coli ) and efflux-deficient (AbΔ3, PΔ3, ΔTolC, and BtΔ2) cells. The heat scale is the base-2 logarithm of the change in MICs. The identified clusters of antibiotics are numbered. (B) Principal-component analysis of antibiotic clusters with best-fit Gaussian distributions. Clusters are numbered as described for panel A. (C) Sizes [log(MW)] and hydrophobicities (logD) of antibiotics belonging to the indicated clusters and fitted to the Gaussian distributions as indicated.

    Techniques Used: Mutagenesis

    Intracellular uptake of the DNA-binding drug HT in different species and their hyperporinated and efflux-deficient derivatives. Data represent real-time kinetics of HT (6 μM final concentration) uptake into B. thailandensis (A), P. aeruginosa PAO1 (B), A. baumannii ATCC 17978 (C), and B. cepacia ATCC 25416 (D). In all panels, blue represents parent strains, orange represents hyperporinated parent strains, gray represents efflux-deficient mutants, and yellow represents efflux-deficient hyperporinated strains. Values in parentheses represent MICs (in milligrams per liter). Error bars represent standard deviations (SD) ( n = 3). AU, arbitrary units.
    Figure Legend Snippet: Intracellular uptake of the DNA-binding drug HT in different species and their hyperporinated and efflux-deficient derivatives. Data represent real-time kinetics of HT (6 μM final concentration) uptake into B. thailandensis (A), P. aeruginosa PAO1 (B), A. baumannii ATCC 17978 (C), and B. cepacia ATCC 25416 (D). In all panels, blue represents parent strains, orange represents hyperporinated parent strains, gray represents efflux-deficient mutants, and yellow represents efflux-deficient hyperporinated strains. Values in parentheses represent MICs (in milligrams per liter). Error bars represent standard deviations (SD) ( n = 3). AU, arbitrary units.

    Techniques Used: Binding Assay, Concentration Assay

    Structures of lipid A moieties and hyperporination of the outer membrane. The results of immunoblotting analyses performed with a monoclonal anti-His antibody, the copy number per cell of the expressed hyperpores, and MICs of vancomycin (VAN) in the induced cells are shown for P. aeruginosa PAO1 (Pae) (A), A. baumannii ATCC 17978 (Abau) (B), and B. thailandensis (Bt) and B. cepacia ATCC 25416 (Bc) (C). M, molecular marker lane.
    Figure Legend Snippet: Structures of lipid A moieties and hyperporination of the outer membrane. The results of immunoblotting analyses performed with a monoclonal anti-His antibody, the copy number per cell of the expressed hyperpores, and MICs of vancomycin (VAN) in the induced cells are shown for P. aeruginosa PAO1 (Pae) (A), A. baumannii ATCC 17978 (Abau) (B), and B. thailandensis (Bt) and B. cepacia ATCC 25416 (Bc) (C). M, molecular marker lane.

    Techniques Used: Marker

    Contributions of the outer membrane barrier and active efflux to susceptibilities of bacteria to antibiotics. (A) A box plot of efflux ratios calculated from drug MICs measured in A. baumannii ATCC 17978, P. aeruginosa PAO1, E. coli , and Bt. Data represent merged ratios of (i) wild-type drug MICs to the drug MICs determined for efflux-deficient strains (i.e., PAO1/PΔ3) and (ii) the drug MICs determined for hyperporinated efflux-plus strains to the drug MICs determined for hyperporinated efflux-minus strains (PAO1-Pore/PΔ3-Pore). The median and the middle half of the distributions of ratios are shown as boxes. (B) Data were determined as described for panel A but for OM ratios calculated from drug MICs measured in A. baumannii ATCC 17978, P. aeruginosa PAO1, E. coli , B. cepacia ATCC 25416, and Bt. Data represent merged ratios of (i) wild-type drug MICs to the drug MICs determined for the hyperporinated strains (i.e., PAO1/PAO1-Pore) and (ii) drug MICs determined for efflux-deficient strains to drug MICs determined for hyperporinated efflux-deficient strains (i.e., PΔ3/PΔ3-Pore).
    Figure Legend Snippet: Contributions of the outer membrane barrier and active efflux to susceptibilities of bacteria to antibiotics. (A) A box plot of efflux ratios calculated from drug MICs measured in A. baumannii ATCC 17978, P. aeruginosa PAO1, E. coli , and Bt. Data represent merged ratios of (i) wild-type drug MICs to the drug MICs determined for efflux-deficient strains (i.e., PAO1/PΔ3) and (ii) the drug MICs determined for hyperporinated efflux-plus strains to the drug MICs determined for hyperporinated efflux-minus strains (PAO1-Pore/PΔ3-Pore). The median and the middle half of the distributions of ratios are shown as boxes. (B) Data were determined as described for panel A but for OM ratios calculated from drug MICs measured in A. baumannii ATCC 17978, P. aeruginosa PAO1, E. coli , B. cepacia ATCC 25416, and Bt. Data represent merged ratios of (i) wild-type drug MICs to the drug MICs determined for the hyperporinated strains (i.e., PAO1/PAO1-Pore) and (ii) drug MICs determined for efflux-deficient strains to drug MICs determined for hyperporinated efflux-deficient strains (i.e., PΔ3/PΔ3-Pore).

    Techniques Used:

    16) Product Images from "Pneumonia infection in mice reveals the involvement of the feoA gene in the pathogenesis of Acinetobacter baumannii"

    Article Title: Pneumonia infection in mice reveals the involvement of the feoA gene in the pathogenesis of Acinetobacter baumannii

    Journal: Virulence

    doi: 10.1080/21505594.2017.1420451

    Pneumonia infection in mice. A) Survival of BALB/c (n = 10 per group) mice after pneumonia infection with A. baumannii ATCC 17978 and Δ0242 strains. Survival was significantly higher in mice infected with the Δ0242 mutant ( p
    Figure Legend Snippet: Pneumonia infection in mice. A) Survival of BALB/c (n = 10 per group) mice after pneumonia infection with A. baumannii ATCC 17978 and Δ0242 strains. Survival was significantly higher in mice infected with the Δ0242 mutant ( p

    Techniques Used: Infection, Mouse Assay, Mutagenesis

    Quantification of bacterial adhesion to A549 cells by the A. baumannii ATCC 17978 strain, the mutant derivative strain Δ0242, the mutant derivative strain Δ3850 and the double mutant strain Δ0242/Δ3850.
    Figure Legend Snippet: Quantification of bacterial adhesion to A549 cells by the A. baumannii ATCC 17978 strain, the mutant derivative strain Δ0242, the mutant derivative strain Δ3850 and the double mutant strain Δ0242/Δ3850.

    Techniques Used: Mutagenesis

    cDNA amplification of genes from the A1S_0242–0244 operon of A. baumannii ATCC 17978 strain. The intergenic regions from genes A1S_0242–0243 and A1S_0243–0244 are shown in lanes 8 and 9, respectively. The intergenic regions from genes A1S_0241–0242 and A1S_0244–0245 are shown in lanes 7 and 10, respectively (negative controls). Genomic DNA was used as template for positive control (lanes 1 to 5, respectively). Lanes 5 and 11 show the gyrB amplification from DNA and cDNA, respectively (positive controls). Lane 6 shows GeneRuler 1 Kb Plus DNA Ladder (Thermo Fisher Scientific).
    Figure Legend Snippet: cDNA amplification of genes from the A1S_0242–0244 operon of A. baumannii ATCC 17978 strain. The intergenic regions from genes A1S_0242–0243 and A1S_0243–0244 are shown in lanes 8 and 9, respectively. The intergenic regions from genes A1S_0241–0242 and A1S_0244–0245 are shown in lanes 7 and 10, respectively (negative controls). Genomic DNA was used as template for positive control (lanes 1 to 5, respectively). Lanes 5 and 11 show the gyrB amplification from DNA and cDNA, respectively (positive controls). Lane 6 shows GeneRuler 1 Kb Plus DNA Ladder (Thermo Fisher Scientific).

    Techniques Used: Amplification, Positive Control

    Quantification of biofilm formation by the A. baumannii ATCC 17978 strain, the mutant derivative strain Δ0242, the mutant derivative strain Δ3850, the double mutant derivative strain Δ0242/Δ3850, the ATCC 17978 harboring the empty vector pWH1266-Km (ATCC 17978 + pWH1266-Km), the mutant derivative strain harboring the empty vector pWH1266-Km (Δ0242 + pWH1266-Km) and the mutant derivative Δ0242 over-expressing the A1S_0242 gene from the pWH1266-Km plasmid (Δ0242 complemented).
    Figure Legend Snippet: Quantification of biofilm formation by the A. baumannii ATCC 17978 strain, the mutant derivative strain Δ0242, the mutant derivative strain Δ3850, the double mutant derivative strain Δ0242/Δ3850, the ATCC 17978 harboring the empty vector pWH1266-Km (ATCC 17978 + pWH1266-Km), the mutant derivative strain harboring the empty vector pWH1266-Km (Δ0242 + pWH1266-Km) and the mutant derivative Δ0242 over-expressing the A1S_0242 gene from the pWH1266-Km plasmid (Δ0242 complemented).

    Techniques Used: Mutagenesis, Plasmid Preparation, Expressing

    Survival of Galleria mellonella larvae (n = 10 per group) after infection with A. baumannii ATCC 17978, Δ0242, Δ3850 and Δ0242/Δ3850 strains. Survival was significantly higher in caterpillars infected with the Δ0242 mutant than those infected with the wild type strain ( p
    Figure Legend Snippet: Survival of Galleria mellonella larvae (n = 10 per group) after infection with A. baumannii ATCC 17978, Δ0242, Δ3850 and Δ0242/Δ3850 strains. Survival was significantly higher in caterpillars infected with the Δ0242 mutant than those infected with the wild type strain ( p

    Techniques Used: Infection, Mutagenesis

    17) Product Images from "Pneumonia infection in mice reveals the involvement of the feoA gene in the pathogenesis of Acinetobacter baumannii"

    Article Title: Pneumonia infection in mice reveals the involvement of the feoA gene in the pathogenesis of Acinetobacter baumannii

    Journal: Virulence

    doi: 10.1080/21505594.2017.1420451

    Pneumonia infection in mice. A) Survival of BALB/c (n = 10 per group) mice after pneumonia infection with A. baumannii ATCC 17978 and Δ0242 strains. Survival was significantly higher in mice infected with the Δ0242 mutant ( p
    Figure Legend Snippet: Pneumonia infection in mice. A) Survival of BALB/c (n = 10 per group) mice after pneumonia infection with A. baumannii ATCC 17978 and Δ0242 strains. Survival was significantly higher in mice infected with the Δ0242 mutant ( p

    Techniques Used: Infection, Mouse Assay, Mutagenesis

    Quantification of bacterial adhesion to A549 cells by the A. baumannii ATCC 17978 strain, the mutant derivative strain Δ0242, the mutant derivative strain Δ3850 and the double mutant strain Δ0242/Δ3850.
    Figure Legend Snippet: Quantification of bacterial adhesion to A549 cells by the A. baumannii ATCC 17978 strain, the mutant derivative strain Δ0242, the mutant derivative strain Δ3850 and the double mutant strain Δ0242/Δ3850.

    Techniques Used: Mutagenesis

    cDNA amplification of genes from the A1S_0242–0244 operon of A. baumannii ATCC 17978 strain. The intergenic regions from genes A1S_0242–0243 and A1S_0243–0244 are shown in lanes 8 and 9, respectively. The intergenic regions from genes A1S_0241–0242 and A1S_0244–0245 are shown in lanes 7 and 10, respectively (negative controls). Genomic DNA was used as template for positive control (lanes 1 to 5, respectively). Lanes 5 and 11 show the gyrB amplification from DNA and cDNA, respectively (positive controls). Lane 6 shows GeneRuler 1 Kb Plus DNA Ladder (Thermo Fisher Scientific).
    Figure Legend Snippet: cDNA amplification of genes from the A1S_0242–0244 operon of A. baumannii ATCC 17978 strain. The intergenic regions from genes A1S_0242–0243 and A1S_0243–0244 are shown in lanes 8 and 9, respectively. The intergenic regions from genes A1S_0241–0242 and A1S_0244–0245 are shown in lanes 7 and 10, respectively (negative controls). Genomic DNA was used as template for positive control (lanes 1 to 5, respectively). Lanes 5 and 11 show the gyrB amplification from DNA and cDNA, respectively (positive controls). Lane 6 shows GeneRuler 1 Kb Plus DNA Ladder (Thermo Fisher Scientific).

    Techniques Used: Amplification, Positive Control

    Quantification of biofilm formation by the A. baumannii ATCC 17978 strain, the mutant derivative strain Δ0242, the mutant derivative strain Δ3850, the double mutant derivative strain Δ0242/Δ3850, the ATCC 17978 harboring the empty vector pWH1266-Km (ATCC 17978 + pWH1266-Km), the mutant derivative strain harboring the empty vector pWH1266-Km (Δ0242 + pWH1266-Km) and the mutant derivative Δ0242 over-expressing the A1S_0242 gene from the pWH1266-Km plasmid (Δ0242 complemented).
    Figure Legend Snippet: Quantification of biofilm formation by the A. baumannii ATCC 17978 strain, the mutant derivative strain Δ0242, the mutant derivative strain Δ3850, the double mutant derivative strain Δ0242/Δ3850, the ATCC 17978 harboring the empty vector pWH1266-Km (ATCC 17978 + pWH1266-Km), the mutant derivative strain harboring the empty vector pWH1266-Km (Δ0242 + pWH1266-Km) and the mutant derivative Δ0242 over-expressing the A1S_0242 gene from the pWH1266-Km plasmid (Δ0242 complemented).

    Techniques Used: Mutagenesis, Plasmid Preparation, Expressing

    Survival of Galleria mellonella larvae (n = 10 per group) after infection with A. baumannii ATCC 17978, Δ0242, Δ3850 and Δ0242/Δ3850 strains. Survival was significantly higher in caterpillars infected with the Δ0242 mutant than those infected with the wild type strain ( p
    Figure Legend Snippet: Survival of Galleria mellonella larvae (n = 10 per group) after infection with A. baumannii ATCC 17978, Δ0242, Δ3850 and Δ0242/Δ3850 strains. Survival was significantly higher in caterpillars infected with the Δ0242 mutant than those infected with the wild type strain ( p

    Techniques Used: Infection, Mutagenesis

    18) Product Images from "The primary transcriptome, small RNAs and regulation of antimicrobial resistance in Acinetobacter baumannii ATCC 17978"

    Article Title: The primary transcriptome, small RNAs and regulation of antimicrobial resistance in Acinetobacter baumannii ATCC 17978

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gky603

    Chromosomal location of coding sequences, small RNAs and TSS of A. baumannii ATCC 17978-mff. Coding sequences are depicted in blue and red (plus and minus strand), small RNAs in light blue (plus strand) and light red (minus strand) and TSS in outer dark grey ring: TSS on the plus strand, inner light gray ring: TSS on the minus strand). The figure was created using Circa (OMGenomics, http://omgenomics.com/circa/ ).
    Figure Legend Snippet: Chromosomal location of coding sequences, small RNAs and TSS of A. baumannii ATCC 17978-mff. Coding sequences are depicted in blue and red (plus and minus strand), small RNAs in light blue (plus strand) and light red (minus strand) and TSS in outer dark grey ring: TSS on the plus strand, inner light gray ring: TSS on the minus strand). The figure was created using Circa (OMGenomics, http://omgenomics.com/circa/ ).

    Techniques Used:

    sRNA conservation in representative members of the order Pseudomonadales. Genomes of multiple Acinetobacter species, Pseudomonas species, and Moraxella catarrhalis were compared using GLSEARCH. The colour scale shows the percentage sequence identity of the 110 sRNAs compared to the reference sequence from A. baumannii ATCC 17978.
    Figure Legend Snippet: sRNA conservation in representative members of the order Pseudomonadales. Genomes of multiple Acinetobacter species, Pseudomonas species, and Moraxella catarrhalis were compared using GLSEARCH. The colour scale shows the percentage sequence identity of the 110 sRNAs compared to the reference sequence from A. baumannii ATCC 17978.

    Techniques Used: Sequencing

    sRNA in A. baumannii ATCC 17978. ( A ) Normalized, mapped sequence reads from RNA-seq show the expression of sRNAs 17, 37, 75, 76, 77, 84, 99 and 100 (yellow arrows). Curved arrows depict TSS identified in this study and lollipop structures are predicted rho-independent terminators. Northern blotting of selected sRNAs are shown to the right. RNA was isolated from ESP and five μg of total RNA was loaded per lane. The sRNA sizes below the individual blots have been predicted from dRNA-seq data. ( B ) Sequence alignment of Group I and Group III sRNAs created with the Geneious Software (v. 8.1.8); colored bases indicate conservation in at least 50% of aligned sequences (A, red; C, blue; G, yellow; T, green). The riboprobes used in Northern blotting are depicted as black bars atop the sRNA alignments.
    Figure Legend Snippet: sRNA in A. baumannii ATCC 17978. ( A ) Normalized, mapped sequence reads from RNA-seq show the expression of sRNAs 17, 37, 75, 76, 77, 84, 99 and 100 (yellow arrows). Curved arrows depict TSS identified in this study and lollipop structures are predicted rho-independent terminators. Northern blotting of selected sRNAs are shown to the right. RNA was isolated from ESP and five μg of total RNA was loaded per lane. The sRNA sizes below the individual blots have been predicted from dRNA-seq data. ( B ) Sequence alignment of Group I and Group III sRNAs created with the Geneious Software (v. 8.1.8); colored bases indicate conservation in at least 50% of aligned sequences (A, red; C, blue; G, yellow; T, green). The riboprobes used in Northern blotting are depicted as black bars atop the sRNA alignments.

    Techniques Used: Sequencing, RNA Sequencing Assay, Expressing, Northern Blot, Isolation, End-sequence Profiling, Software

    Characterization of TSS. (A) Schematic explaining TSS categories. P: Primary TSS, S: Secondary TSS, I: Internal TSS, A: Antisense TSS, O: Orphan TSS. (B) Pie chart showing the number of TSS per category. (C) Histogram showing the number and length of 5′ UTRs of primary (red) and secondary (black) TSS. The inset illustrates the frequency of occurrence of nucleotides around the TSS. (D) Meme-derived motifs showing −35 and −10 region of A. baumannii ATCC 17978 promoters.
    Figure Legend Snippet: Characterization of TSS. (A) Schematic explaining TSS categories. P: Primary TSS, S: Secondary TSS, I: Internal TSS, A: Antisense TSS, O: Orphan TSS. (B) Pie chart showing the number of TSS per category. (C) Histogram showing the number and length of 5′ UTRs of primary (red) and secondary (black) TSS. The inset illustrates the frequency of occurrence of nucleotides around the TSS. (D) Meme-derived motifs showing −35 and −10 region of A. baumannii ATCC 17978 promoters.

    Techniques Used: Derivative Assay

    19) Product Images from "2-Hydroxylation of Acinetobacter baumannii Lipid A Contributes to Virulence"

    Article Title: 2-Hydroxylation of Acinetobacter baumannii Lipid A Contributes to Virulence

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00066-19

    Deletion of lpxO decreases A. baumannii resistance to CAMPs. Shown is percent survival of A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn7 lpxO ) following 1 h of exposure to polymyxin B (A), colistin (B), and human β-defensin 3 (C). Values are presented as the means ± standard deviations (SD) from three independent experiments measured in duplicate. P values were
    Figure Legend Snippet: Deletion of lpxO decreases A. baumannii resistance to CAMPs. Shown is percent survival of A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn7 lpxO ) following 1 h of exposure to polymyxin B (A), colistin (B), and human β-defensin 3 (C). Values are presented as the means ± standard deviations (SD) from three independent experiments measured in duplicate. P values were

    Techniques Used: Mutagenesis

    lpxO deletion in A. baumannii results in upregulation of inflammatory responses in macrophages upon infection. (A) tnf α expression in iBMDMs infected for 5 h with UV-killed A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ) by reverse transcriptase quantitative real‐time PCR. Values are presented as the means ± SD from three independent cDNA preparations measured in duplicate. (B) TNF-α secretion by iBMDMs stimulated for 5 h with UV-killed A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ). In panels A and B, P values were
    Figure Legend Snippet: lpxO deletion in A. baumannii results in upregulation of inflammatory responses in macrophages upon infection. (A) tnf α expression in iBMDMs infected for 5 h with UV-killed A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ) by reverse transcriptase quantitative real‐time PCR. Values are presented as the means ± SD from three independent cDNA preparations measured in duplicate. (B) TNF-α secretion by iBMDMs stimulated for 5 h with UV-killed A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ). In panels A and B, P values were

    Techniques Used: Infection, Expressing, Mutagenesis, Real-time Polymerase Chain Reaction

    Deletion of lpxO increases human phagocyte-mediated killing of A. baumannii . Three hundred μl of fresh human blood (from three different donors) was mixed with 1 × 10 7 CFU of A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ) and incubated at 37°C for 3 h. The bacterial counts recovered were then divided by the initial counts. Experiments were performed with duplicate samples on three independent occasions. *, P
    Figure Legend Snippet: Deletion of lpxO increases human phagocyte-mediated killing of A. baumannii . Three hundred μl of fresh human blood (from three different donors) was mixed with 1 × 10 7 CFU of A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ) and incubated at 37°C for 3 h. The bacterial counts recovered were then divided by the initial counts. Experiments were performed with duplicate samples on three independent occasions. *, P

    Techniques Used: Mutagenesis, Incubation

    A. baumannii LpxO 2-hydroxylates lipid A. (A) A. baumannii encodes an LpxO homolog with 62% and 59% sequence similarity to K. pneumoniae and S. enterica serovar Typhimurium LpxO, respectively. Shown are negative-ion MALDI-TOF mass spectrometry spectra of lipid A purified from A. baumannii ATCC 17978 (B), A. baumannii Δ lpxO mutant (Δ lpxO ) (C), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ) (D). Data represent the mass-to-charge ratios ( m/z ) of each lipid A species detected and are representative of three extractions. (E) Proposed lipid A structures of the species produced by wild-type A. baumannii based on previous publications ( 7 – 11 ).
    Figure Legend Snippet: A. baumannii LpxO 2-hydroxylates lipid A. (A) A. baumannii encodes an LpxO homolog with 62% and 59% sequence similarity to K. pneumoniae and S. enterica serovar Typhimurium LpxO, respectively. Shown are negative-ion MALDI-TOF mass spectrometry spectra of lipid A purified from A. baumannii ATCC 17978 (B), A. baumannii Δ lpxO mutant (Δ lpxO ) (C), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ) (D). Data represent the mass-to-charge ratios ( m/z ) of each lipid A species detected and are representative of three extractions. (E) Proposed lipid A structures of the species produced by wild-type A. baumannii based on previous publications ( 7 – 11 ).

    Techniques Used: Sequencing, Mass Spectrometry, Purification, Mutagenesis, Produced

    lpxO deletion in A. baumannii results in a decrease in il10 expression. (A) il10 expression in iBMDMs infected for 5 h with UV-killed A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn7 lpxO mutant (Δ lpxO ::Tn7 lpxO ) by reverse transcriptase quantitative real‐time PCR. Values are presented as the means ± SD from three independent cDNA preparations measured in duplicate. (B) tnf α expression in il10 −/− iBMDMs infected for 5 h with UV-killed A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn7 lpxO mutant (Δ lpxO ::Tn7 lpxO ) by reverse transcriptase quantitative real‐time PCR. Values are presented as the means ± SD from two independent cDNA preparations measured in duplicate. (C) il10 expression in control (AS) and CREB siRNA-transfected iBMDMs infected for 5 h with UV-killed A. baumannii ATCC 17978 by reverse transcriptase quantitative real‐time PCR. Values are presented as the means ± SD from two independent cDNA preparations measured in duplicate. (D) Immunoblot analysis of phosphor-CREB (pCREB) and tubulin levels in lysates of iBMDMs infected with UV-killed A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn7 lpxO mutant (Δ lpxO ::Tn7 lpxO ). Data are representative of at least three independent experiments. In panels A to C, P values were
    Figure Legend Snippet: lpxO deletion in A. baumannii results in a decrease in il10 expression. (A) il10 expression in iBMDMs infected for 5 h with UV-killed A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn7 lpxO mutant (Δ lpxO ::Tn7 lpxO ) by reverse transcriptase quantitative real‐time PCR. Values are presented as the means ± SD from three independent cDNA preparations measured in duplicate. (B) tnf α expression in il10 −/− iBMDMs infected for 5 h with UV-killed A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn7 lpxO mutant (Δ lpxO ::Tn7 lpxO ) by reverse transcriptase quantitative real‐time PCR. Values are presented as the means ± SD from two independent cDNA preparations measured in duplicate. (C) il10 expression in control (AS) and CREB siRNA-transfected iBMDMs infected for 5 h with UV-killed A. baumannii ATCC 17978 by reverse transcriptase quantitative real‐time PCR. Values are presented as the means ± SD from two independent cDNA preparations measured in duplicate. (D) Immunoblot analysis of phosphor-CREB (pCREB) and tubulin levels in lysates of iBMDMs infected with UV-killed A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn7 lpxO mutant (Δ lpxO ::Tn7 lpxO ). Data are representative of at least three independent experiments. In panels A to C, P values were

    Techniques Used: Expressing, Infection, Mutagenesis, Real-time Polymerase Chain Reaction, Transfection

    A. baumannii lpxO mutant displays decreased virulence in the G. mellonella waxworm infection model. (A) Percent survival of G. mellonella over 72 h postinfection with 5 × 10 4 organisms of A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ). Thirty larvae were infected in each group. Level of significance was determined using the log-rank (Mantel-Cox) test with Bonferroni correction for multiple comparisons. (B) Percent survival of A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ) following 1 h of exposure to G. mellonella hemolymph obtained from larvae challenged with heat-killed E. coli . (C to E) G. mellonella antimicrobial peptide expression determined after 12 h of infection with A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ) by reverse transcriptase quantitative real‐time PCR. Three larvae per group were infected, and values are presented as the means ± SD from two independent cDNA preparations measured in duplicate. In panels B to E, P values were
    Figure Legend Snippet: A. baumannii lpxO mutant displays decreased virulence in the G. mellonella waxworm infection model. (A) Percent survival of G. mellonella over 72 h postinfection with 5 × 10 4 organisms of A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ). Thirty larvae were infected in each group. Level of significance was determined using the log-rank (Mantel-Cox) test with Bonferroni correction for multiple comparisons. (B) Percent survival of A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ) following 1 h of exposure to G. mellonella hemolymph obtained from larvae challenged with heat-killed E. coli . (C to E) G. mellonella antimicrobial peptide expression determined after 12 h of infection with A. baumannii ATCC 17978, A. baumannii Δ lpxO mutant (Δ lpxO ), and A. baumannii Δ lpxO ::Tn 7lpxO mutant (Δ lpxO ::Tn 7lpxO ) by reverse transcriptase quantitative real‐time PCR. Three larvae per group were infected, and values are presented as the means ± SD from two independent cDNA preparations measured in duplicate. In panels B to E, P values were

    Techniques Used: Mutagenesis, Infection, Expressing, Real-time Polymerase Chain Reaction

    20) Product Images from "Antibiotic Resistance Acquired through a DNA Damage-Inducible Response in Acinetobacter baumannii"

    Article Title: Antibiotic Resistance Acquired through a DNA Damage-Inducible Response in Acinetobacter baumannii

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.02176-12

    Intracellular concentrations of A. baumannii ATCC 17978 DNA damage-inducible proteins increase upon UV irradiation. At 160 J m −2 , there was 40-fold more RecA protein, 2.5-fold more UvrA, and 3-fold more DinB than for the untreated samples, while RpoB remained constant. A. baumannii cultures were grown to exponential phase, as indicated in Materials and Methods, and irradiated with increasing amounts of UV (J m −2 ). Equal amounts of whole-cell lysates per treatment were probed with polyclonal anti-RecA, polyclonal anti-UvrA, polyclonal anti-DinB, and monoclonal anti-RpoB antibodies (refer to Materials and Methods). Antibodies used were raised against the E. coli proteins. A comparative experiment using the isogenic recA ::Km strain could not be performed due to its extreme sensitivity to UV irradiation.
    Figure Legend Snippet: Intracellular concentrations of A. baumannii ATCC 17978 DNA damage-inducible proteins increase upon UV irradiation. At 160 J m −2 , there was 40-fold more RecA protein, 2.5-fold more UvrA, and 3-fold more DinB than for the untreated samples, while RpoB remained constant. A. baumannii cultures were grown to exponential phase, as indicated in Materials and Methods, and irradiated with increasing amounts of UV (J m −2 ). Equal amounts of whole-cell lysates per treatment were probed with polyclonal anti-RecA, polyclonal anti-UvrA, polyclonal anti-DinB, and monoclonal anti-RpoB antibodies (refer to Materials and Methods). Antibodies used were raised against the E. coli proteins. A comparative experiment using the isogenic recA ::Km strain could not be performed due to its extreme sensitivity to UV irradiation.

    Techniques Used: Irradiation

    The A. baumannii ATCC 17978 predicted umuC and umuD genes are organized differently in A. baumannii ATCC 17978 than they are in E. coli . (A) There is one umuDC operon in the E. coli (Ec) chromosome in which the umuD ORF is expressed approximately 10-fold better than umuC ). This frameshift in the gene is depicted as overlapping arrows. (B) A. baumannii ATCC 17978 (Ab) has two putative umuDC operons in an organization similar to the one in E. coli , but within the umuDC (0636 to 0637) operon there is an overlap between the umuD and umuC genes of 20 nucleotides (depicted by overlapping arrows). In the umuDC (1174 to 1173) operon, we find no overlap between the two predicted genes. There are also two unlinked predicted umuC genes and one unlinked predicted umuD gene. For easier identification, locus tags (“A1S_” is not included before the numbers) are included as part of each A. baumannii gene name. Arrows represent predicted ORFs, and white boxes represent promoter (P) or putative promoter (P*) regions.
    Figure Legend Snippet: The A. baumannii ATCC 17978 predicted umuC and umuD genes are organized differently in A. baumannii ATCC 17978 than they are in E. coli . (A) There is one umuDC operon in the E. coli (Ec) chromosome in which the umuD ORF is expressed approximately 10-fold better than umuC ). This frameshift in the gene is depicted as overlapping arrows. (B) A. baumannii ATCC 17978 (Ab) has two putative umuDC operons in an organization similar to the one in E. coli , but within the umuDC (0636 to 0637) operon there is an overlap between the umuD and umuC genes of 20 nucleotides (depicted by overlapping arrows). In the umuDC (1174 to 1173) operon, we find no overlap between the two predicted genes. There are also two unlinked predicted umuC genes and one unlinked predicted umuD gene. For easier identification, locus tags (“A1S_” is not included before the numbers) are included as part of each A. baumannii gene name. Arrows represent predicted ORFs, and white boxes represent promoter (P) or putative promoter (P*) regions.

    Techniques Used:

    Representative, evolutionarily conserved DNA damage response genes are expressed in A. baumannii ATCC 17978. The predicted genes encoding DNA damage response genes are all expressed in the recA + strain, though at different levels. The relative expression of each gene is shown as a percentage of the level of expression of 16S rRNA, a standard housekeeping gene. In the recA ::Km strain, most genes analyzed had no detectable change in relative basal-level gene expression. Some genes showed modest detectable decreases and modest to moderate increases in expression, which suggests a role for RecA in gene regulation. Semiquantitative RT-PCR was performed on total RNA purified from untreated cultures of A. baumannii ATCC 17978. See Materials and Methods for details of this experimental procedure. Gene-specific RT-PCR primers were used to amplify approximately 300 bp of either the unique junctions between the umuD and umuC genes organized as operons or unique sequences of the unlinked genes. Locus tags from the A. baumannii ATCC 17978 genome (“A1S_” is not included before the numbers) are included as part of the umuD and umuC names. Data from a representative experiment are shown.
    Figure Legend Snippet: Representative, evolutionarily conserved DNA damage response genes are expressed in A. baumannii ATCC 17978. The predicted genes encoding DNA damage response genes are all expressed in the recA + strain, though at different levels. The relative expression of each gene is shown as a percentage of the level of expression of 16S rRNA, a standard housekeeping gene. In the recA ::Km strain, most genes analyzed had no detectable change in relative basal-level gene expression. Some genes showed modest detectable decreases and modest to moderate increases in expression, which suggests a role for RecA in gene regulation. Semiquantitative RT-PCR was performed on total RNA purified from untreated cultures of A. baumannii ATCC 17978. See Materials and Methods for details of this experimental procedure. Gene-specific RT-PCR primers were used to amplify approximately 300 bp of either the unique junctions between the umuD and umuC genes organized as operons or unique sequences of the unlinked genes. Locus tags from the A. baumannii ATCC 17978 genome (“A1S_” is not included before the numbers) are included as part of the umuD and umuC names. Data from a representative experiment are shown.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Purification

    21) Product Images from "Antibiotic Resistance Acquired through a DNA Damage-Inducible Response in Acinetobacter baumannii"

    Article Title: Antibiotic Resistance Acquired through a DNA Damage-Inducible Response in Acinetobacter baumannii

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.02176-12

    Intracellular concentrations of A. baumannii ATCC 17978 DNA damage-inducible proteins increase upon UV irradiation. At 160 J m −2 , there was 40-fold more RecA protein, 2.5-fold more UvrA, and 3-fold more DinB than for the untreated samples, while RpoB remained constant. A. baumannii cultures were grown to exponential phase, as indicated in Materials and Methods, and irradiated with increasing amounts of UV (J m −2 ). Equal amounts of whole-cell lysates per treatment were probed with polyclonal anti-RecA, polyclonal anti-UvrA, polyclonal anti-DinB, and monoclonal anti-RpoB antibodies (refer to Materials and Methods). Antibodies used were raised against the E. coli proteins. A comparative experiment using the isogenic recA ::Km strain could not be performed due to its extreme sensitivity to UV irradiation.
    Figure Legend Snippet: Intracellular concentrations of A. baumannii ATCC 17978 DNA damage-inducible proteins increase upon UV irradiation. At 160 J m −2 , there was 40-fold more RecA protein, 2.5-fold more UvrA, and 3-fold more DinB than for the untreated samples, while RpoB remained constant. A. baumannii cultures were grown to exponential phase, as indicated in Materials and Methods, and irradiated with increasing amounts of UV (J m −2 ). Equal amounts of whole-cell lysates per treatment were probed with polyclonal anti-RecA, polyclonal anti-UvrA, polyclonal anti-DinB, and monoclonal anti-RpoB antibodies (refer to Materials and Methods). Antibodies used were raised against the E. coli proteins. A comparative experiment using the isogenic recA ::Km strain could not be performed due to its extreme sensitivity to UV irradiation.

    Techniques Used: Irradiation

    The A. baumannii ATCC 17978 predicted umuC and umuD genes are organized differently in A. baumannii ATCC 17978 than they are in E. coli . (A) There is one umuDC operon in the E. coli (Ec) chromosome in which the umuD ORF is expressed approximately 10-fold better than umuC ). This frameshift in the gene is depicted as overlapping arrows. (B) A. baumannii ATCC 17978 (Ab) has two putative umuDC operons in an organization similar to the one in E. coli , but within the umuDC (0636 to 0637) operon there is an overlap between the umuD and umuC genes of 20 nucleotides (depicted by overlapping arrows). In the umuDC (1174 to 1173) operon, we find no overlap between the two predicted genes. There are also two unlinked predicted umuC genes and one unlinked predicted umuD gene. For easier identification, locus tags (“A1S_” is not included before the numbers) are included as part of each A. baumannii gene name. Arrows represent predicted ORFs, and white boxes represent promoter (P) or putative promoter (P*) regions.
    Figure Legend Snippet: The A. baumannii ATCC 17978 predicted umuC and umuD genes are organized differently in A. baumannii ATCC 17978 than they are in E. coli . (A) There is one umuDC operon in the E. coli (Ec) chromosome in which the umuD ORF is expressed approximately 10-fold better than umuC ). This frameshift in the gene is depicted as overlapping arrows. (B) A. baumannii ATCC 17978 (Ab) has two putative umuDC operons in an organization similar to the one in E. coli , but within the umuDC (0636 to 0637) operon there is an overlap between the umuD and umuC genes of 20 nucleotides (depicted by overlapping arrows). In the umuDC (1174 to 1173) operon, we find no overlap between the two predicted genes. There are also two unlinked predicted umuC genes and one unlinked predicted umuD gene. For easier identification, locus tags (“A1S_” is not included before the numbers) are included as part of each A. baumannii gene name. Arrows represent predicted ORFs, and white boxes represent promoter (P) or putative promoter (P*) regions.

    Techniques Used:

    Representative, evolutionarily conserved DNA damage response genes are expressed in A. baumannii ATCC 17978. The predicted genes encoding DNA damage response genes are all expressed in the recA + strain, though at different levels. The relative expression of each gene is shown as a percentage of the level of expression of 16S rRNA, a standard housekeeping gene. In the recA ::Km strain, most genes analyzed had no detectable change in relative basal-level gene expression. Some genes showed modest detectable decreases and modest to moderate increases in expression, which suggests a role for RecA in gene regulation. Semiquantitative RT-PCR was performed on total RNA purified from untreated cultures of A. baumannii ATCC 17978. See Materials and Methods for details of this experimental procedure. Gene-specific RT-PCR primers were used to amplify approximately 300 bp of either the unique junctions between the umuD and umuC genes organized as operons or unique sequences of the unlinked genes. Locus tags from the A. baumannii ATCC 17978 genome (“A1S_” is not included before the numbers) are included as part of the umuD and umuC names. Data from a representative experiment are shown.
    Figure Legend Snippet: Representative, evolutionarily conserved DNA damage response genes are expressed in A. baumannii ATCC 17978. The predicted genes encoding DNA damage response genes are all expressed in the recA + strain, though at different levels. The relative expression of each gene is shown as a percentage of the level of expression of 16S rRNA, a standard housekeeping gene. In the recA ::Km strain, most genes analyzed had no detectable change in relative basal-level gene expression. Some genes showed modest detectable decreases and modest to moderate increases in expression, which suggests a role for RecA in gene regulation. Semiquantitative RT-PCR was performed on total RNA purified from untreated cultures of A. baumannii ATCC 17978. See Materials and Methods for details of this experimental procedure. Gene-specific RT-PCR primers were used to amplify approximately 300 bp of either the unique junctions between the umuD and umuC genes organized as operons or unique sequences of the unlinked genes. Locus tags from the A. baumannii ATCC 17978 genome (“A1S_” is not included before the numbers) are included as part of the umuD and umuC names. Data from a representative experiment are shown.

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Purification

    22) Product Images from "Horizontal Gene Transfer and Assortative Recombination within the Acinetobacter baumannii Clinical Population Provide Genetic Diversity at the Single carO Gene, Encoding a Major Outer Membrane Protein Channel ▿ Gene, Encoding a Major Outer Membrane Protein Channel ▿ † Gene, Encoding a Major Outer Membrane Protein Channel ▿ † ‡"

    Article Title: Horizontal Gene Transfer and Assortative Recombination within the Acinetobacter baumannii Clinical Population Provide Genetic Diversity at the Single carO Gene, Encoding a Major Outer Membrane Protein Channel ▿ Gene, Encoding a Major Outer Membrane Protein Channel ▿ † Gene, Encoding a Major Outer Membrane Protein Channel ▿ † ‡

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.01533-10

    Characterization of A. baumannii ATCC 17978 ΔcarO mutant cells separately expressing the different plasmid-carried carO variants. (A) Growth curves on l -ornithine as the sole carbon source of A. baumannii ATCC 17978 ΔcarO cells transformed
    Figure Legend Snippet: Characterization of A. baumannii ATCC 17978 ΔcarO mutant cells separately expressing the different plasmid-carried carO variants. (A) Growth curves on l -ornithine as the sole carbon source of A. baumannii ATCC 17978 ΔcarO cells transformed

    Techniques Used: Mutagenesis, Expressing, Plasmid Preparation, Transformation Assay

    23) Product Images from "Simple Method for Markerless Gene Deletion in Multidrug-Resistant Acinetobacter baumannii"

    Article Title: Simple Method for Markerless Gene Deletion in Multidrug-Resistant Acinetobacter baumannii

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.03975-14

    Gene deletion in A. baumannii strains other than ATCC 19606. (A) The susceptibilities of A. baumannii strains ATCC 17978 and 1656-2 to increasing amounts of antibiotics were explored by measuring their growth after 12 h on LB medium containing different concentrations of antibiotics. Asterisks indicate significant differences ( P
    Figure Legend Snippet: Gene deletion in A. baumannii strains other than ATCC 19606. (A) The susceptibilities of A. baumannii strains ATCC 17978 and 1656-2 to increasing amounts of antibiotics were explored by measuring their growth after 12 h on LB medium containing different concentrations of antibiotics. Asterisks indicate significant differences ( P

    Techniques Used:

    24) Product Images from "The Opportunistic Human Pathogen Acinetobacter baumannii Senses and Responds to Light ▿ Senses and Responds to Light ▿ †"

    Article Title: The Opportunistic Human Pathogen Acinetobacter baumannii Senses and Responds to Light ▿ Senses and Responds to Light ▿ †

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00917-10

    Killing of tup1 mutant C. albicans by A. baumannii cells. (A) Fungal filaments were incubated in sterile medium or medium inoculated with bacterial cells from the ATCC 17978 parental strain or the BlsA-deficient derivative ATCC 17978.OR either in the presence of blue light (L) or in darkness (D). (B) Experiment similar to that shown in panel A but using the ATCC 17978.ORc and ATCC 17978.ORp derivatives harboring the complementing plasmid pWHBLSA and the empty vector pWH1266, respectively. Error bars show the standard error of the mean.
    Figure Legend Snippet: Killing of tup1 mutant C. albicans by A. baumannii cells. (A) Fungal filaments were incubated in sterile medium or medium inoculated with bacterial cells from the ATCC 17978 parental strain or the BlsA-deficient derivative ATCC 17978.OR either in the presence of blue light (L) or in darkness (D). (B) Experiment similar to that shown in panel A but using the ATCC 17978.ORc and ATCC 17978.ORp derivatives harboring the complementing plasmid pWHBLSA and the empty vector pWH1266, respectively. Error bars show the standard error of the mean.

    Techniques Used: Mutagenesis, Incubation, Plasmid Preparation

    Effects of light and temperature on blsA transcript levels. RNA from A. baumannii ATCC 17978 cells grown overnight on motility plates at 24°C and 37°C in the presence of blue light (L) or in darkness (D) was used as the template for qRT-PCR using blsA -specific primers. Transcription of recA was used as a constitutively expressed internal control. Error bars show the standard error of the mean.
    Figure Legend Snippet: Effects of light and temperature on blsA transcript levels. RNA from A. baumannii ATCC 17978 cells grown overnight on motility plates at 24°C and 37°C in the presence of blue light (L) or in darkness (D) was used as the template for qRT-PCR using blsA -specific primers. Transcription of recA was used as a constitutively expressed internal control. Error bars show the standard error of the mean.

    Techniques Used: Quantitative RT-PCR

    25) Product Images from "The Opportunistic Human Pathogen Acinetobacter baumannii Senses and Responds to Light ▿ Senses and Responds to Light ▿ †"

    Article Title: The Opportunistic Human Pathogen Acinetobacter baumannii Senses and Responds to Light ▿ Senses and Responds to Light ▿ †

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00917-10

    Killing of tup1 mutant C. albicans by A. baumannii cells. (A) Fungal filaments were incubated in sterile medium or medium inoculated with bacterial cells from the ATCC 17978 parental strain or the BlsA-deficient derivative ATCC 17978.OR either in the presence of blue light (L) or in darkness (D). (B) Experiment similar to that shown in panel A but using the ATCC 17978.ORc and ATCC 17978.ORp derivatives harboring the complementing plasmid pWHBLSA and the empty vector pWH1266, respectively. Error bars show the standard error of the mean.
    Figure Legend Snippet: Killing of tup1 mutant C. albicans by A. baumannii cells. (A) Fungal filaments were incubated in sterile medium or medium inoculated with bacterial cells from the ATCC 17978 parental strain or the BlsA-deficient derivative ATCC 17978.OR either in the presence of blue light (L) or in darkness (D). (B) Experiment similar to that shown in panel A but using the ATCC 17978.ORc and ATCC 17978.ORp derivatives harboring the complementing plasmid pWHBLSA and the empty vector pWH1266, respectively. Error bars show the standard error of the mean.

    Techniques Used: Mutagenesis, Incubation, Plasmid Preparation

    Effects of light and temperature on blsA transcript levels. RNA from A. baumannii ATCC 17978 cells grown overnight on motility plates at 24°C and 37°C in the presence of blue light (L) or in darkness (D) was used as the template for qRT-PCR using blsA -specific primers. Transcription of recA was used as a constitutively expressed internal control. Error bars show the standard error of the mean.
    Figure Legend Snippet: Effects of light and temperature on blsA transcript levels. RNA from A. baumannii ATCC 17978 cells grown overnight on motility plates at 24°C and 37°C in the presence of blue light (L) or in darkness (D) was used as the template for qRT-PCR using blsA -specific primers. Transcription of recA was used as a constitutively expressed internal control. Error bars show the standard error of the mean.

    Techniques Used: Quantitative RT-PCR

    26) Product Images from "The Acinetobacter baumannii Omp33-36 Porin Is a Virulence Factor That Induces Apoptosis and Modulates Autophagy in Human Cells"

    Article Title: The Acinetobacter baumannii Omp33-36 Porin Is a Virulence Factor That Induces Apoptosis and Modulates Autophagy in Human Cells

    Journal: Infection and Immunity

    doi: 10.1128/IAI.02034-14

    (A to D) Autophagosomes (double-membrane structures) with A. baumannii cells replicating within them. (A) TEM of RAW 264.7 macrophages incubated for 12 h with A. baumannii ATCC 17978; (B) group of intact bacteria enclosed by double membranes in RAW 264.7
    Figure Legend Snippet: (A to D) Autophagosomes (double-membrane structures) with A. baumannii cells replicating within them. (A) TEM of RAW 264.7 macrophages incubated for 12 h with A. baumannii ATCC 17978; (B) group of intact bacteria enclosed by double membranes in RAW 264.7

    Techniques Used: Transmission Electron Microscopy, Incubation

    (A) Blockage of autophagy by OMVs carrying porins (with and without Omp33-36). p62 and LC3B levels of OMVs from A. baumannii ATCC 17978 and A. baumannii ATCCΔ mapA analyzed by Western blotting. *, densitometry of p62 and LC3B-II at 0 and 24 h from
    Figure Legend Snippet: (A) Blockage of autophagy by OMVs carrying porins (with and without Omp33-36). p62 and LC3B levels of OMVs from A. baumannii ATCC 17978 and A. baumannii ATCCΔ mapA analyzed by Western blotting. *, densitometry of p62 and LC3B-II at 0 and 24 h from

    Techniques Used: Western Blot

    (A) Greater intracellular persistence of A. baumannii ATCC 17978 (carrying Omp33-36). The numbers of CFU (indicated on the y axis) recovered after infection with A. baumannii ATCC 17978 and A. baumannii ATCCΔ mapA for 12 h and 16 h are shown. (B)
    Figure Legend Snippet: (A) Greater intracellular persistence of A. baumannii ATCC 17978 (carrying Omp33-36). The numbers of CFU (indicated on the y axis) recovered after infection with A. baumannii ATCC 17978 and A. baumannii ATCCΔ mapA for 12 h and 16 h are shown. (B)

    Techniques Used: Infection

    27) Product Images from "The Acinetobacter baumannii Omp33-36 Porin Is a Virulence Factor That Induces Apoptosis and Modulates Autophagy in Human Cells"

    Article Title: The Acinetobacter baumannii Omp33-36 Porin Is a Virulence Factor That Induces Apoptosis and Modulates Autophagy in Human Cells

    Journal: Infection and Immunity

    doi: 10.1128/IAI.02034-14

    (A to D) Autophagosomes (double-membrane structures) with A. baumannii cells replicating within them. (A) TEM of RAW 264.7 macrophages incubated for 12 h with A. baumannii ATCC 17978; (B) group of intact bacteria enclosed by double membranes in RAW 264.7
    Figure Legend Snippet: (A to D) Autophagosomes (double-membrane structures) with A. baumannii cells replicating within them. (A) TEM of RAW 264.7 macrophages incubated for 12 h with A. baumannii ATCC 17978; (B) group of intact bacteria enclosed by double membranes in RAW 264.7

    Techniques Used: Transmission Electron Microscopy, Incubation

    (A) Blockage of autophagy by OMVs carrying porins (with and without Omp33-36). p62 and LC3B levels of OMVs from A. baumannii ATCC 17978 and A. baumannii ATCCΔ mapA analyzed by Western blotting. *, densitometry of p62 and LC3B-II at 0 and 24 h from
    Figure Legend Snippet: (A) Blockage of autophagy by OMVs carrying porins (with and without Omp33-36). p62 and LC3B levels of OMVs from A. baumannii ATCC 17978 and A. baumannii ATCCΔ mapA analyzed by Western blotting. *, densitometry of p62 and LC3B-II at 0 and 24 h from

    Techniques Used: Western Blot

    (A) Greater intracellular persistence of A. baumannii ATCC 17978 (carrying Omp33-36). The numbers of CFU (indicated on the y axis) recovered after infection with A. baumannii ATCC 17978 and A. baumannii ATCCΔ mapA for 12 h and 16 h are shown. (B)
    Figure Legend Snippet: (A) Greater intracellular persistence of A. baumannii ATCC 17978 (carrying Omp33-36). The numbers of CFU (indicated on the y axis) recovered after infection with A. baumannii ATCC 17978 and A. baumannii ATCCΔ mapA for 12 h and 16 h are shown. (B)

    Techniques Used: Infection

    28) Product Images from "Identification of Ata, a Multifunctional Trimeric Autotransporter of Acinetobacter baumannii"

    Article Title: Identification of Ata, a Multifunctional Trimeric Autotransporter of Acinetobacter baumannii

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.06769-11

    Quantification of Ata expression in A. baumannii ATCC 17978 by flow cytometry during various growth phases. A. baumannii was grown in LB to early exponential (OD 650 of 0.025 or 0.1), mid-exponential (OD 650 of 0.4), late exponential (OD 650 of 0.8), or stationary phase (OD 650 of 1.2), labeled with rabbit antibody to Ata and secondary goat antibody to rabbit IgG conjugated to Alexa 488 fluorescent dye, and analyzed by flow cytometry. Results represent the mean fluorescent intensity (MFI) of 500,000 cells, and bars indicate the averages from three independent experiments ± standard errors of the means (SEM).
    Figure Legend Snippet: Quantification of Ata expression in A. baumannii ATCC 17978 by flow cytometry during various growth phases. A. baumannii was grown in LB to early exponential (OD 650 of 0.025 or 0.1), mid-exponential (OD 650 of 0.4), late exponential (OD 650 of 0.8), or stationary phase (OD 650 of 1.2), labeled with rabbit antibody to Ata and secondary goat antibody to rabbit IgG conjugated to Alexa 488 fluorescent dye, and analyzed by flow cytometry. Results represent the mean fluorescent intensity (MFI) of 500,000 cells, and bars indicate the averages from three independent experiments ± standard errors of the means (SEM).

    Techniques Used: Expressing, Flow Cytometry, Cytometry, Labeling

    Survival curves of mice ( n = 8; C57BL/6) following intraperitoneal infection with A. baumannii ATCC 17978 (1.6 × 10 7 CFU/mouse), Δ ata (1.6 × 10 7 CFU/mouse), Δ ata -pLVB-Ata ( ata gene with its native promoter cloned into pBAD18Kan-Ori; 1.4 × 10 7 CFU/mouse), and Δ ata -pBAD18Kan-Ori (empty vector pBAD18Kan-Ori; 1.2 × 10 7 CFU/mouse). P = 0.0005 by log-rank test in a Kaplan-Meier analysis of wild-type ATCC 17978 versus Δ ata and ATCC 17978 versus Δ ata -pBAD18kan-Ori.
    Figure Legend Snippet: Survival curves of mice ( n = 8; C57BL/6) following intraperitoneal infection with A. baumannii ATCC 17978 (1.6 × 10 7 CFU/mouse), Δ ata (1.6 × 10 7 CFU/mouse), Δ ata -pLVB-Ata ( ata gene with its native promoter cloned into pBAD18Kan-Ori; 1.4 × 10 7 CFU/mouse), and Δ ata -pBAD18Kan-Ori (empty vector pBAD18Kan-Ori; 1.2 × 10 7 CFU/mouse). P = 0.0005 by log-rank test in a Kaplan-Meier analysis of wild-type ATCC 17978 versus Δ ata and ATCC 17978 versus Δ ata -pBAD18kan-Ori.

    Techniques Used: Mouse Assay, Infection, Clone Assay, Plasmid Preparation

    Quantitative biofilm formation by A. baumannii ATCC 17978, Δ ata , and Δ ata -c on polystyrene surfaces. A. baumannii 17978 and Δ ata were grown in LB and ATCC 17978 Δ ata -c in LB plus 2% arabinose or 0.2% glucose under static conditions for 24 h at 37°C. Total biofilm formation (OD 595 ) was normalized by bacterial growth (OD 650 ). The bars indicate the means of 15 tubes from five independent experiments ± SEM. P values were determined by one-way ANOVA with Tukey's post hoc analysis. ns, not significant.
    Figure Legend Snippet: Quantitative biofilm formation by A. baumannii ATCC 17978, Δ ata , and Δ ata -c on polystyrene surfaces. A. baumannii 17978 and Δ ata were grown in LB and ATCC 17978 Δ ata -c in LB plus 2% arabinose or 0.2% glucose under static conditions for 24 h at 37°C. Total biofilm formation (OD 595 ) was normalized by bacterial growth (OD 650 ). The bars indicate the means of 15 tubes from five independent experiments ± SEM. P values were determined by one-way ANOVA with Tukey's post hoc analysis. ns, not significant.

    Techniques Used:

    (A) Immunoblot of Ata-V5-His fusion protein expressed in E. coli . Outer membrane proteins (OMPs) of E. coli LMG194 harboring pAta-V5-6×His were prepared after induction with either 0.2% glucose (lane 1) or 0.2% arabinose (lane 2) and then tested by Western blotting with a monoclonal antibody (MAb) directed against the V5 peptide. Samples from arabinose-induced OMPs were treated with 70% formic acid and detected as before (lane 3). The predicted monomeric mass of Ata-V5-His fusion proteins is 13.8 kDa. Migration distances of molecular mass markers (in kDa) are indicated on the right. (B) Detection of Ata-V5-His fusion protein in E. coli pAta-V5-6×His by CLSM. E. coli pAta-V5-6×His cultures were grown in M9 plus 0.25% Casamino Acids and induced with 0.2% arabinose or 0.2% glucose. The V5 epitope was labeled with a mouse anti-V5 MAb and a secondary goat anti-mouse Alexa 488 fluorescent antibody (green channel), and cell nucleic acids were stained with DAPI (blue channel). (a) E. coli pAta-V5-6×His induced with arabinose (green channel) or (b) glucose (green channel). Also shown are DAPI-stained nuclei of E. coli cells (blue channel) after induction with arabinose (c) or glucose (d). (C) CLSM detection of Ata in A. baumannii ATCC 17978, Δ ata , and Δ ata -c strains. A. baumannii ATCC 17978, Δ ata , and Δ ata -c induced with arabinose or glucose were labeled with rabbit anti-Ata antisera and a secondary goat anti-rabbit Alexa 488 fluorescent antibody. Nucleic acids were stained with DAPI. Merged images of Alexa 488- and DAPI-stained A. baumannii ATCC 17978 (a) and Δ ata (b), Δ ata -c induced with glucose (c), and Δ ata -c induced with arabinose (d) are shown.
    Figure Legend Snippet: (A) Immunoblot of Ata-V5-His fusion protein expressed in E. coli . Outer membrane proteins (OMPs) of E. coli LMG194 harboring pAta-V5-6×His were prepared after induction with either 0.2% glucose (lane 1) or 0.2% arabinose (lane 2) and then tested by Western blotting with a monoclonal antibody (MAb) directed against the V5 peptide. Samples from arabinose-induced OMPs were treated with 70% formic acid and detected as before (lane 3). The predicted monomeric mass of Ata-V5-His fusion proteins is 13.8 kDa. Migration distances of molecular mass markers (in kDa) are indicated on the right. (B) Detection of Ata-V5-His fusion protein in E. coli pAta-V5-6×His by CLSM. E. coli pAta-V5-6×His cultures were grown in M9 plus 0.25% Casamino Acids and induced with 0.2% arabinose or 0.2% glucose. The V5 epitope was labeled with a mouse anti-V5 MAb and a secondary goat anti-mouse Alexa 488 fluorescent antibody (green channel), and cell nucleic acids were stained with DAPI (blue channel). (a) E. coli pAta-V5-6×His induced with arabinose (green channel) or (b) glucose (green channel). Also shown are DAPI-stained nuclei of E. coli cells (blue channel) after induction with arabinose (c) or glucose (d). (C) CLSM detection of Ata in A. baumannii ATCC 17978, Δ ata , and Δ ata -c strains. A. baumannii ATCC 17978, Δ ata , and Δ ata -c induced with arabinose or glucose were labeled with rabbit anti-Ata antisera and a secondary goat anti-rabbit Alexa 488 fluorescent antibody. Nucleic acids were stained with DAPI. Merged images of Alexa 488- and DAPI-stained A. baumannii ATCC 17978 (a) and Δ ata (b), Δ ata -c induced with glucose (c), and Δ ata -c induced with arabinose (d) are shown.

    Techniques Used: Western Blot, Migration, Confocal Laser Scanning Microscopy, Labeling, Staining

    Adhesive properties of Ata. (A) Dose-dependent binding of Ata to selected ECM/BM components. ECM/BM proteins, including collagen types I, II, III, IV, and V, heparan, fibronectin, vitronectin, and laminin, and BSA (as a control) were used at 5 μg/well, and binding of Ata was quantified by ELISA. Data points represent the means from four independent experiments ± SEM. (B) Binding of A. baumannii ATCC 17978, Δ ata , and Δ ata -c induced with 2% arabinose or 0.2% glucose to immobilized collagen type IV. The bars indicate the means from three independent experiments ± SEM. P values were determined by one-way ANOVA with Tukey's post hoc analysis. ns, not significant.
    Figure Legend Snippet: Adhesive properties of Ata. (A) Dose-dependent binding of Ata to selected ECM/BM components. ECM/BM proteins, including collagen types I, II, III, IV, and V, heparan, fibronectin, vitronectin, and laminin, and BSA (as a control) were used at 5 μg/well, and binding of Ata was quantified by ELISA. Data points represent the means from four independent experiments ± SEM. (B) Binding of A. baumannii ATCC 17978, Δ ata , and Δ ata -c induced with 2% arabinose or 0.2% glucose to immobilized collagen type IV. The bars indicate the means from three independent experiments ± SEM. P values were determined by one-way ANOVA with Tukey's post hoc analysis. ns, not significant.

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay

    Analysis of Ata expression among A. baumannii clinical isolates. (A) Flow-cytometric analysis of Ata production among A. baumannii clinical isolates positive for the ata gene by PCR. A. baumannii strains were grown in LB and probed with rabbit anti-Ata antibodies and a secondary goat anti-rabbit Alexa 488 fluorescent antibody. Approximately 500,000 cells were then analyzed by flow cytometry, and antibody binding was expressed as mean fluorescent intensity (MFI). Bars represent the fold ratio (in MFI) of each A. baumannii clinical isolate to the MFI of the ATCC 17978 Δ ata strain. (B) Western blot (WB) analysis of Ata levels in 4 strains producing high levels of Ata (ATCC 17978, S19, I30, and N10) and in 4 strains producing low levels of Ata (I42, I25, I31, and I28) as determined by FACS and the Ata-negative strain ATCC 17978 Δ ata , which was used as a control. Migration distances of molecular mass markers (in kDa) are indicated on the left. (Strains used for WB analysis are highlighted with arrows in panel A). Outer membrane proteins were extracted, resolved by SDS-PAGE, and transferred to a PVDF membrane, and Ata was detected by WB with anti-Ata rabbit antibodies and a secondary goat anti-rabbit IgG.
    Figure Legend Snippet: Analysis of Ata expression among A. baumannii clinical isolates. (A) Flow-cytometric analysis of Ata production among A. baumannii clinical isolates positive for the ata gene by PCR. A. baumannii strains were grown in LB and probed with rabbit anti-Ata antibodies and a secondary goat anti-rabbit Alexa 488 fluorescent antibody. Approximately 500,000 cells were then analyzed by flow cytometry, and antibody binding was expressed as mean fluorescent intensity (MFI). Bars represent the fold ratio (in MFI) of each A. baumannii clinical isolate to the MFI of the ATCC 17978 Δ ata strain. (B) Western blot (WB) analysis of Ata levels in 4 strains producing high levels of Ata (ATCC 17978, S19, I30, and N10) and in 4 strains producing low levels of Ata (I42, I25, I31, and I28) as determined by FACS and the Ata-negative strain ATCC 17978 Δ ata , which was used as a control. Migration distances of molecular mass markers (in kDa) are indicated on the left. (Strains used for WB analysis are highlighted with arrows in panel A). Outer membrane proteins were extracted, resolved by SDS-PAGE, and transferred to a PVDF membrane, and Ata was detected by WB with anti-Ata rabbit antibodies and a secondary goat anti-rabbit IgG.

    Techniques Used: Expressing, Flow Cytometry, Polymerase Chain Reaction, Cytometry, Binding Assay, Western Blot, FACS, Migration, SDS Page

    29) Product Images from "Identification of Ata, a Multifunctional Trimeric Autotransporter of Acinetobacter baumannii"

    Article Title: Identification of Ata, a Multifunctional Trimeric Autotransporter of Acinetobacter baumannii

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.06769-11

    Quantification of Ata expression in A. baumannii ATCC 17978 by flow cytometry during various growth phases. A. baumannii was grown in LB to early exponential (OD 650 of 0.025 or 0.1), mid-exponential (OD 650 of 0.4), late exponential (OD 650 of 0.8), or stationary phase (OD 650 of 1.2), labeled with rabbit antibody to Ata and secondary goat antibody to rabbit IgG conjugated to Alexa 488 fluorescent dye, and analyzed by flow cytometry. Results represent the mean fluorescent intensity (MFI) of 500,000 cells, and bars indicate the averages from three independent experiments ± standard errors of the means (SEM).
    Figure Legend Snippet: Quantification of Ata expression in A. baumannii ATCC 17978 by flow cytometry during various growth phases. A. baumannii was grown in LB to early exponential (OD 650 of 0.025 or 0.1), mid-exponential (OD 650 of 0.4), late exponential (OD 650 of 0.8), or stationary phase (OD 650 of 1.2), labeled with rabbit antibody to Ata and secondary goat antibody to rabbit IgG conjugated to Alexa 488 fluorescent dye, and analyzed by flow cytometry. Results represent the mean fluorescent intensity (MFI) of 500,000 cells, and bars indicate the averages from three independent experiments ± standard errors of the means (SEM).

    Techniques Used: Expressing, Flow Cytometry, Cytometry, Labeling

    Survival curves of mice ( n = 8; C57BL/6) following intraperitoneal infection with A. baumannii ATCC 17978 (1.6 × 10 7 CFU/mouse), Δ ata (1.6 × 10 7 CFU/mouse), Δ ata -pLVB-Ata ( ata gene with its native promoter cloned into pBAD18Kan-Ori; 1.4 × 10 7 CFU/mouse), and Δ ata -pBAD18Kan-Ori (empty vector pBAD18Kan-Ori; 1.2 × 10 7 CFU/mouse). P = 0.0005 by log-rank test in a Kaplan-Meier analysis of wild-type ATCC 17978 versus Δ ata and ATCC 17978 versus Δ ata -pBAD18kan-Ori.
    Figure Legend Snippet: Survival curves of mice ( n = 8; C57BL/6) following intraperitoneal infection with A. baumannii ATCC 17978 (1.6 × 10 7 CFU/mouse), Δ ata (1.6 × 10 7 CFU/mouse), Δ ata -pLVB-Ata ( ata gene with its native promoter cloned into pBAD18Kan-Ori; 1.4 × 10 7 CFU/mouse), and Δ ata -pBAD18Kan-Ori (empty vector pBAD18Kan-Ori; 1.2 × 10 7 CFU/mouse). P = 0.0005 by log-rank test in a Kaplan-Meier analysis of wild-type ATCC 17978 versus Δ ata and ATCC 17978 versus Δ ata -pBAD18kan-Ori.

    Techniques Used: Mouse Assay, Infection, Clone Assay, Plasmid Preparation

    Quantitative biofilm formation by A. baumannii ATCC 17978, Δ ata , and Δ ata -c on polystyrene surfaces. A. baumannii 17978 and Δ ata were grown in LB and ATCC 17978 Δ ata -c in LB plus 2% arabinose or 0.2% glucose under static conditions for 24 h at 37°C. Total biofilm formation (OD 595 ) was normalized by bacterial growth (OD 650 ). The bars indicate the means of 15 tubes from five independent experiments ± SEM. P values were determined by one-way ANOVA with Tukey's post hoc analysis. ns, not significant.
    Figure Legend Snippet: Quantitative biofilm formation by A. baumannii ATCC 17978, Δ ata , and Δ ata -c on polystyrene surfaces. A. baumannii 17978 and Δ ata were grown in LB and ATCC 17978 Δ ata -c in LB plus 2% arabinose or 0.2% glucose under static conditions for 24 h at 37°C. Total biofilm formation (OD 595 ) was normalized by bacterial growth (OD 650 ). The bars indicate the means of 15 tubes from five independent experiments ± SEM. P values were determined by one-way ANOVA with Tukey's post hoc analysis. ns, not significant.

    Techniques Used:

    (A) Immunoblot of Ata-V5-His fusion protein expressed in E. coli . Outer membrane proteins (OMPs) of E. coli LMG194 harboring pAta-V5-6×His were prepared after induction with either 0.2% glucose (lane 1) or 0.2% arabinose (lane 2) and then tested by Western blotting with a monoclonal antibody (MAb) directed against the V5 peptide. Samples from arabinose-induced OMPs were treated with 70% formic acid and detected as before (lane 3). The predicted monomeric mass of Ata-V5-His fusion proteins is 13.8 kDa. Migration distances of molecular mass markers (in kDa) are indicated on the right. (B) Detection of Ata-V5-His fusion protein in E. coli pAta-V5-6×His by CLSM. E. coli pAta-V5-6×His cultures were grown in M9 plus 0.25% Casamino Acids and induced with 0.2% arabinose or 0.2% glucose. The V5 epitope was labeled with a mouse anti-V5 MAb and a secondary goat anti-mouse Alexa 488 fluorescent antibody (green channel), and cell nucleic acids were stained with DAPI (blue channel). (a) E. coli pAta-V5-6×His induced with arabinose (green channel) or (b) glucose (green channel). Also shown are DAPI-stained nuclei of E. coli cells (blue channel) after induction with arabinose (c) or glucose (d). (C) CLSM detection of Ata in A. baumannii ATCC 17978, Δ ata , and Δ ata -c strains. A. baumannii ATCC 17978, Δ ata , and Δ ata -c induced with arabinose or glucose were labeled with rabbit anti-Ata antisera and a secondary goat anti-rabbit Alexa 488 fluorescent antibody. Nucleic acids were stained with DAPI. Merged images of Alexa 488- and DAPI-stained A. baumannii ATCC 17978 (a) and Δ ata (b), Δ ata -c induced with glucose (c), and Δ ata -c induced with arabinose (d) are shown.
    Figure Legend Snippet: (A) Immunoblot of Ata-V5-His fusion protein expressed in E. coli . Outer membrane proteins (OMPs) of E. coli LMG194 harboring pAta-V5-6×His were prepared after induction with either 0.2% glucose (lane 1) or 0.2% arabinose (lane 2) and then tested by Western blotting with a monoclonal antibody (MAb) directed against the V5 peptide. Samples from arabinose-induced OMPs were treated with 70% formic acid and detected as before (lane 3). The predicted monomeric mass of Ata-V5-His fusion proteins is 13.8 kDa. Migration distances of molecular mass markers (in kDa) are indicated on the right. (B) Detection of Ata-V5-His fusion protein in E. coli pAta-V5-6×His by CLSM. E. coli pAta-V5-6×His cultures were grown in M9 plus 0.25% Casamino Acids and induced with 0.2% arabinose or 0.2% glucose. The V5 epitope was labeled with a mouse anti-V5 MAb and a secondary goat anti-mouse Alexa 488 fluorescent antibody (green channel), and cell nucleic acids were stained with DAPI (blue channel). (a) E. coli pAta-V5-6×His induced with arabinose (green channel) or (b) glucose (green channel). Also shown are DAPI-stained nuclei of E. coli cells (blue channel) after induction with arabinose (c) or glucose (d). (C) CLSM detection of Ata in A. baumannii ATCC 17978, Δ ata , and Δ ata -c strains. A. baumannii ATCC 17978, Δ ata , and Δ ata -c induced with arabinose or glucose were labeled with rabbit anti-Ata antisera and a secondary goat anti-rabbit Alexa 488 fluorescent antibody. Nucleic acids were stained with DAPI. Merged images of Alexa 488- and DAPI-stained A. baumannii ATCC 17978 (a) and Δ ata (b), Δ ata -c induced with glucose (c), and Δ ata -c induced with arabinose (d) are shown.

    Techniques Used: Western Blot, Migration, Confocal Laser Scanning Microscopy, Labeling, Staining

    Adhesive properties of Ata. (A) Dose-dependent binding of Ata to selected ECM/BM components. ECM/BM proteins, including collagen types I, II, III, IV, and V, heparan, fibronectin, vitronectin, and laminin, and BSA (as a control) were used at 5 μg/well, and binding of Ata was quantified by ELISA. Data points represent the means from four independent experiments ± SEM. (B) Binding of A. baumannii ATCC 17978, Δ ata , and Δ ata -c induced with 2% arabinose or 0.2% glucose to immobilized collagen type IV. The bars indicate the means from three independent experiments ± SEM. P values were determined by one-way ANOVA with Tukey's post hoc analysis. ns, not significant.
    Figure Legend Snippet: Adhesive properties of Ata. (A) Dose-dependent binding of Ata to selected ECM/BM components. ECM/BM proteins, including collagen types I, II, III, IV, and V, heparan, fibronectin, vitronectin, and laminin, and BSA (as a control) were used at 5 μg/well, and binding of Ata was quantified by ELISA. Data points represent the means from four independent experiments ± SEM. (B) Binding of A. baumannii ATCC 17978, Δ ata , and Δ ata -c induced with 2% arabinose or 0.2% glucose to immobilized collagen type IV. The bars indicate the means from three independent experiments ± SEM. P values were determined by one-way ANOVA with Tukey's post hoc analysis. ns, not significant.

    Techniques Used: Binding Assay, Enzyme-linked Immunosorbent Assay

    Analysis of Ata expression among A. baumannii clinical isolates. (A) Flow-cytometric analysis of Ata production among A. baumannii clinical isolates positive for the ata gene by PCR. A. baumannii strains were grown in LB and probed with rabbit anti-Ata antibodies and a secondary goat anti-rabbit Alexa 488 fluorescent antibody. Approximately 500,000 cells were then analyzed by flow cytometry, and antibody binding was expressed as mean fluorescent intensity (MFI). Bars represent the fold ratio (in MFI) of each A. baumannii clinical isolate to the MFI of the ATCC 17978 Δ ata strain. (B) Western blot (WB) analysis of Ata levels in 4 strains producing high levels of Ata (ATCC 17978, S19, I30, and N10) and in 4 strains producing low levels of Ata (I42, I25, I31, and I28) as determined by FACS and the Ata-negative strain ATCC 17978 Δ ata , which was used as a control. Migration distances of molecular mass markers (in kDa) are indicated on the left. (Strains used for WB analysis are highlighted with arrows in panel A). Outer membrane proteins were extracted, resolved by SDS-PAGE, and transferred to a PVDF membrane, and Ata was detected by WB with anti-Ata rabbit antibodies and a secondary goat anti-rabbit IgG.
    Figure Legend Snippet: Analysis of Ata expression among A. baumannii clinical isolates. (A) Flow-cytometric analysis of Ata production among A. baumannii clinical isolates positive for the ata gene by PCR. A. baumannii strains were grown in LB and probed with rabbit anti-Ata antibodies and a secondary goat anti-rabbit Alexa 488 fluorescent antibody. Approximately 500,000 cells were then analyzed by flow cytometry, and antibody binding was expressed as mean fluorescent intensity (MFI). Bars represent the fold ratio (in MFI) of each A. baumannii clinical isolate to the MFI of the ATCC 17978 Δ ata strain. (B) Western blot (WB) analysis of Ata levels in 4 strains producing high levels of Ata (ATCC 17978, S19, I30, and N10) and in 4 strains producing low levels of Ata (I42, I25, I31, and I28) as determined by FACS and the Ata-negative strain ATCC 17978 Δ ata , which was used as a control. Migration distances of molecular mass markers (in kDa) are indicated on the left. (Strains used for WB analysis are highlighted with arrows in panel A). Outer membrane proteins were extracted, resolved by SDS-PAGE, and transferred to a PVDF membrane, and Ata was detected by WB with anti-Ata rabbit antibodies and a secondary goat anti-rabbit IgG.

    Techniques Used: Expressing, Flow Cytometry, Polymerase Chain Reaction, Cytometry, Binding Assay, Western Blot, FACS, Migration, SDS Page

    30) Product Images from "Triclosan Can Select for an AdeIJK-Overexpressing Mutant of Acinetobacter baumannii ATCC 17978 That Displays Reduced Susceptibility to Multiple Antibiotics"

    Article Title: Triclosan Can Select for an AdeIJK-Overexpressing Mutant of Acinetobacter baumannii ATCC 17978 That Displays Reduced Susceptibility to Multiple Antibiotics

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.03074-14

    Expression of adeJ in Acinetobacter baumannii ATCC 17978 complemented with adeN . Expression of the adeJ was measured by qRT-PCR in Acinetobacter baumannii ATCC 17978 complemented with the wild-type adeN . We used 16S rRNA as the housekeeping gene control.
    Figure Legend Snippet: Expression of adeJ in Acinetobacter baumannii ATCC 17978 complemented with adeN . Expression of the adeJ was measured by qRT-PCR in Acinetobacter baumannii ATCC 17978 complemented with the wild-type adeN . We used 16S rRNA as the housekeeping gene control.

    Techniques Used: Expressing, Quantitative RT-PCR

    31) Product Images from "The biochemical origins of the surface enhanced Raman spectra of bacteria: metabolomics profiling by SERS"

    Article Title: The biochemical origins of the surface enhanced Raman spectra of bacteria: metabolomics profiling by SERS

    Journal: Analytical and bioanalytical chemistry

    doi: 10.1007/s00216-016-9540-x

    Comparison of SERS spectra of three bacterial species and some model compounds. The SERS spectra of S. aureus (NCTC 8325), E. faecalis (ATCC 29212) and A baumannii (ATCC 17978) are compared with adenine, hypoxanthine and xanthine SERS spectra, respectively.
    Figure Legend Snippet: Comparison of SERS spectra of three bacterial species and some model compounds. The SERS spectra of S. aureus (NCTC 8325), E. faecalis (ATCC 29212) and A baumannii (ATCC 17978) are compared with adenine, hypoxanthine and xanthine SERS spectra, respectively.

    Techniques Used:

    32) Product Images from "Impact of a Cross-Kingdom Signaling Molecule of Candida albicans on Acinetobacter baumannii Physiology"

    Article Title: Impact of a Cross-Kingdom Signaling Molecule of Candida albicans on Acinetobacter baumannii Physiology

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01540-15

    Drug efflux pumps are required for farnesol resistance in A. baumannii ATCC 17978. Bacteria were cultured overnight in either HI medium containing either farnesol (0.5 mM), the efflux pump inhibitor phenyl-arginine-β-naphthylamide dihydrochloride
    Figure Legend Snippet: Drug efflux pumps are required for farnesol resistance in A. baumannii ATCC 17978. Bacteria were cultured overnight in either HI medium containing either farnesol (0.5 mM), the efflux pump inhibitor phenyl-arginine-β-naphthylamide dihydrochloride

    Techniques Used: Cell Culture

    Impact of farnesol on susceptibility to the antibiotic colistin. A. baumannii strains ATCC 17978 (A) and AB0059 (B) were grown in HI medium alone (control) or HI medium containing either a subinhibitory concentration of farnesol (0.5 mM), colistin (1
    Figure Legend Snippet: Impact of farnesol on susceptibility to the antibiotic colistin. A. baumannii strains ATCC 17978 (A) and AB0059 (B) were grown in HI medium alone (control) or HI medium containing either a subinhibitory concentration of farnesol (0.5 mM), colistin (1

    Techniques Used: Concentration Assay

    Impact of farnesol on various phenotypes of A. baumannii ATCC 17978. (A) Cellular morphology in the presence of farnesol. Overnight cultures of bacteria grown in the presence or absence of farnesol were fixed and viewed by scanning electron microscopy.
    Figure Legend Snippet: Impact of farnesol on various phenotypes of A. baumannii ATCC 17978. (A) Cellular morphology in the presence of farnesol. Overnight cultures of bacteria grown in the presence or absence of farnesol were fixed and viewed by scanning electron microscopy.

    Techniques Used: Electron Microscopy

    33) Product Images from "Impact of a Cross-Kingdom Signaling Molecule of Candida albicans on Acinetobacter baumannii Physiology"

    Article Title: Impact of a Cross-Kingdom Signaling Molecule of Candida albicans on Acinetobacter baumannii Physiology

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01540-15

    Drug efflux pumps are required for farnesol resistance in A. baumannii ATCC 17978. Bacteria were cultured overnight in either HI medium containing either farnesol (0.5 mM), the efflux pump inhibitor phenyl-arginine-β-naphthylamide dihydrochloride
    Figure Legend Snippet: Drug efflux pumps are required for farnesol resistance in A. baumannii ATCC 17978. Bacteria were cultured overnight in either HI medium containing either farnesol (0.5 mM), the efflux pump inhibitor phenyl-arginine-β-naphthylamide dihydrochloride

    Techniques Used: Cell Culture

    Impact of farnesol on susceptibility to the antibiotic colistin. A. baumannii strains ATCC 17978 (A) and AB0059 (B) were grown in HI medium alone (control) or HI medium containing either a subinhibitory concentration of farnesol (0.5 mM), colistin (1
    Figure Legend Snippet: Impact of farnesol on susceptibility to the antibiotic colistin. A. baumannii strains ATCC 17978 (A) and AB0059 (B) were grown in HI medium alone (control) or HI medium containing either a subinhibitory concentration of farnesol (0.5 mM), colistin (1

    Techniques Used: Concentration Assay

    Impact of farnesol on various phenotypes of A. baumannii ATCC 17978. (A) Cellular morphology in the presence of farnesol. Overnight cultures of bacteria grown in the presence or absence of farnesol were fixed and viewed by scanning electron microscopy.
    Figure Legend Snippet: Impact of farnesol on various phenotypes of A. baumannii ATCC 17978. (A) Cellular morphology in the presence of farnesol. Overnight cultures of bacteria grown in the presence or absence of farnesol were fixed and viewed by scanning electron microscopy.

    Techniques Used: Electron Microscopy

    34) Product Images from "Impact of a Cross-Kingdom Signaling Molecule of Candida albicans on Acinetobacter baumannii Physiology"

    Article Title: Impact of a Cross-Kingdom Signaling Molecule of Candida albicans on Acinetobacter baumannii Physiology

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01540-15

    Drug efflux pumps are required for farnesol resistance in A. baumannii ATCC 17978. Bacteria were cultured overnight in either HI medium containing either farnesol (0.5 mM), the efflux pump inhibitor phenyl-arginine-β-naphthylamide dihydrochloride
    Figure Legend Snippet: Drug efflux pumps are required for farnesol resistance in A. baumannii ATCC 17978. Bacteria were cultured overnight in either HI medium containing either farnesol (0.5 mM), the efflux pump inhibitor phenyl-arginine-β-naphthylamide dihydrochloride

    Techniques Used: Cell Culture

    Impact of farnesol on susceptibility to the antibiotic colistin. A. baumannii strains ATCC 17978 (A) and AB0059 (B) were grown in HI medium alone (control) or HI medium containing either a subinhibitory concentration of farnesol (0.5 mM), colistin (1
    Figure Legend Snippet: Impact of farnesol on susceptibility to the antibiotic colistin. A. baumannii strains ATCC 17978 (A) and AB0059 (B) were grown in HI medium alone (control) or HI medium containing either a subinhibitory concentration of farnesol (0.5 mM), colistin (1

    Techniques Used: Concentration Assay

    Impact of farnesol on various phenotypes of A. baumannii ATCC 17978. (A) Cellular morphology in the presence of farnesol. Overnight cultures of bacteria grown in the presence or absence of farnesol were fixed and viewed by scanning electron microscopy.
    Figure Legend Snippet: Impact of farnesol on various phenotypes of A. baumannii ATCC 17978. (A) Cellular morphology in the presence of farnesol. Overnight cultures of bacteria grown in the presence or absence of farnesol were fixed and viewed by scanning electron microscopy.

    Techniques Used: Electron Microscopy

    35) Product Images from "Subinhibitory Concentrations of Trimethoprim and Sulfamethoxazole Prevent Biofilm Formation by Acinetobacter baumannii through Inhibition of Csu Pilus Expression"

    Article Title: Subinhibitory Concentrations of Trimethoprim and Sulfamethoxazole Prevent Biofilm Formation by Acinetobacter baumannii through Inhibition of Csu Pilus Expression

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00778-17

    Requirement for Csu pili for biofilm formation in A. baumannii ATCC 17978. (A) Representative crystal violet assay image of A. baumannii strains on polystyrene surfaces. The biofilms were stained with crystal violet and dissolved in 10% acetic acid solution. A. baumannii ATCC 17978 wild-type and complemented strains, but not the csuD mutant strain, formed biofilms. The endogenous pAB3 plasmid and the pBAV shuttle vector did not affect biofilm formation. The A. baumannii ATCC 19606 strain was used as a control. (B) Quantitative determination of biofilm formation. To normalize cell growth rates, A. baumannii strains were grown in LB medium and collected at OD 600 values of 1.0 to 1.2. To count CFU, collected cells were plated on LB medium (top); to quantify the biofilms, absorbance was measured at OD 550 (bottom). Points indicate mean values, and error bars indicate standard deviations of triplicate samples. *, P ≤ 0.002, significant reduction of biofilm production in csuD mutant strains, compared to 17978pAB3−.
    Figure Legend Snippet: Requirement for Csu pili for biofilm formation in A. baumannii ATCC 17978. (A) Representative crystal violet assay image of A. baumannii strains on polystyrene surfaces. The biofilms were stained with crystal violet and dissolved in 10% acetic acid solution. A. baumannii ATCC 17978 wild-type and complemented strains, but not the csuD mutant strain, formed biofilms. The endogenous pAB3 plasmid and the pBAV shuttle vector did not affect biofilm formation. The A. baumannii ATCC 19606 strain was used as a control. (B) Quantitative determination of biofilm formation. To normalize cell growth rates, A. baumannii strains were grown in LB medium and collected at OD 600 values of 1.0 to 1.2. To count CFU, collected cells were plated on LB medium (top); to quantify the biofilms, absorbance was measured at OD 550 (bottom). Points indicate mean values, and error bars indicate standard deviations of triplicate samples. *, P ≤ 0.002, significant reduction of biofilm production in csuD mutant strains, compared to 17978pAB3−.

    Techniques Used: Crystal Violet Assay, Staining, Mutagenesis, Plasmid Preparation

    36) Product Images from "Subinhibitory Concentrations of Trimethoprim and Sulfamethoxazole Prevent Biofilm Formation by Acinetobacter baumannii through Inhibition of Csu Pilus Expression"

    Article Title: Subinhibitory Concentrations of Trimethoprim and Sulfamethoxazole Prevent Biofilm Formation by Acinetobacter baumannii through Inhibition of Csu Pilus Expression

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00778-17

    Requirement for Csu pili for biofilm formation in A. baumannii ATCC 17978. (A) Representative crystal violet assay image of A. baumannii strains on polystyrene surfaces. The biofilms were stained with crystal violet and dissolved in 10% acetic acid solution. A. baumannii ATCC 17978 wild-type and complemented strains, but not the csuD mutant strain, formed biofilms. The endogenous pAB3 plasmid and the pBAV shuttle vector did not affect biofilm formation. The A. baumannii ATCC 19606 strain was used as a control. (B) Quantitative determination of biofilm formation. To normalize cell growth rates, A. baumannii strains were grown in LB medium and collected at OD 600 values of 1.0 to 1.2. To count CFU, collected cells were plated on LB medium (top); to quantify the biofilms, absorbance was measured at OD 550 (bottom). Points indicate mean values, and error bars indicate standard deviations of triplicate samples. *, P ≤ 0.002, significant reduction of biofilm production in csuD mutant strains, compared to 17978pAB3−.
    Figure Legend Snippet: Requirement for Csu pili for biofilm formation in A. baumannii ATCC 17978. (A) Representative crystal violet assay image of A. baumannii strains on polystyrene surfaces. The biofilms were stained with crystal violet and dissolved in 10% acetic acid solution. A. baumannii ATCC 17978 wild-type and complemented strains, but not the csuD mutant strain, formed biofilms. The endogenous pAB3 plasmid and the pBAV shuttle vector did not affect biofilm formation. The A. baumannii ATCC 19606 strain was used as a control. (B) Quantitative determination of biofilm formation. To normalize cell growth rates, A. baumannii strains were grown in LB medium and collected at OD 600 values of 1.0 to 1.2. To count CFU, collected cells were plated on LB medium (top); to quantify the biofilms, absorbance was measured at OD 550 (bottom). Points indicate mean values, and error bars indicate standard deviations of triplicate samples. *, P ≤ 0.002, significant reduction of biofilm production in csuD mutant strains, compared to 17978pAB3−.

    Techniques Used: Crystal Violet Assay, Staining, Mutagenesis, Plasmid Preparation

    37) Product Images from "Effect of Incubation Temperature on Antibiotic Resistance and Virulence Factors of Acinetobacter baumannii ATCC 17978"

    Article Title: Effect of Incubation Temperature on Antibiotic Resistance and Virulence Factors of Acinetobacter baumannii ATCC 17978

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01514-17

    Comparison of biofilm formation by A. baumannii ATCC 17978 at 28°C and 37°C. Biofilm formation was significantly greater at 28°C than at 37°C. Statistical analysis was carried out using two-way analysis of variance (ANOVA). *, P = 0.001.
    Figure Legend Snippet: Comparison of biofilm formation by A. baumannii ATCC 17978 at 28°C and 37°C. Biofilm formation was significantly greater at 28°C than at 37°C. Statistical analysis was carried out using two-way analysis of variance (ANOVA). *, P = 0.001.

    Techniques Used:

    Comparison of twitching motility of A. baumannii ATCC 17978 at 28°C and 37°C. (A) The bacteria were able to cover the entire surface of the plate after 18 h of incubation at 37°C, but growth was significantly less at 28°C than at 37°C. Each data point represents a biological replicate. (B) The picture shows representative plates inoculated with A. baumannii ATCC 17978 and incubated at 28°C and 37°C. Statistical analysis was carried out using a paired t test.
    Figure Legend Snippet: Comparison of twitching motility of A. baumannii ATCC 17978 at 28°C and 37°C. (A) The bacteria were able to cover the entire surface of the plate after 18 h of incubation at 37°C, but growth was significantly less at 28°C than at 37°C. Each data point represents a biological replicate. (B) The picture shows representative plates inoculated with A. baumannii ATCC 17978 and incubated at 28°C and 37°C. Statistical analysis was carried out using a paired t test.

    Techniques Used: Incubation

    38) Product Images from "Effect of Incubation Temperature on Antibiotic Resistance and Virulence Factors of Acinetobacter baumannii ATCC 17978"

    Article Title: Effect of Incubation Temperature on Antibiotic Resistance and Virulence Factors of Acinetobacter baumannii ATCC 17978

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01514-17

    Comparison of biofilm formation by A. baumannii ATCC 17978 at 28°C and 37°C. Biofilm formation was significantly greater at 28°C than at 37°C. Statistical analysis was carried out using two-way analysis of variance (ANOVA). *, P = 0.001.
    Figure Legend Snippet: Comparison of biofilm formation by A. baumannii ATCC 17978 at 28°C and 37°C. Biofilm formation was significantly greater at 28°C than at 37°C. Statistical analysis was carried out using two-way analysis of variance (ANOVA). *, P = 0.001.

    Techniques Used:

    Comparison of twitching motility of A. baumannii ATCC 17978 at 28°C and 37°C. (A) The bacteria were able to cover the entire surface of the plate after 18 h of incubation at 37°C, but growth was significantly less at 28°C than at 37°C. Each data point represents a biological replicate. (B) The picture shows representative plates inoculated with A. baumannii ATCC 17978 and incubated at 28°C and 37°C. Statistical analysis was carried out using a paired t test.
    Figure Legend Snippet: Comparison of twitching motility of A. baumannii ATCC 17978 at 28°C and 37°C. (A) The bacteria were able to cover the entire surface of the plate after 18 h of incubation at 37°C, but growth was significantly less at 28°C than at 37°C. Each data point represents a biological replicate. (B) The picture shows representative plates inoculated with A. baumannii ATCC 17978 and incubated at 28°C and 37°C. Statistical analysis was carried out using a paired t test.

    Techniques Used: Incubation

    39) Product Images from "Effect of Incubation Temperature on Antibiotic Resistance and Virulence Factors of Acinetobacter baumannii ATCC 17978"

    Article Title: Effect of Incubation Temperature on Antibiotic Resistance and Virulence Factors of Acinetobacter baumannii ATCC 17978

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01514-17

    Comparison of biofilm formation by A. baumannii ATCC 17978 at 28°C and 37°C. Biofilm formation was significantly greater at 28°C than at 37°C. Statistical analysis was carried out using two-way analysis of variance (ANOVA). *, P = 0.001.
    Figure Legend Snippet: Comparison of biofilm formation by A. baumannii ATCC 17978 at 28°C and 37°C. Biofilm formation was significantly greater at 28°C than at 37°C. Statistical analysis was carried out using two-way analysis of variance (ANOVA). *, P = 0.001.

    Techniques Used:

    Comparison of twitching motility of A. baumannii ATCC 17978 at 28°C and 37°C. (A) The bacteria were able to cover the entire surface of the plate after 18 h of incubation at 37°C, but growth was significantly less at 28°C than at 37°C. Each data point represents a biological replicate. (B) The picture shows representative plates inoculated with A. baumannii ATCC 17978 and incubated at 28°C and 37°C. Statistical analysis was carried out using a paired t test.
    Figure Legend Snippet: Comparison of twitching motility of A. baumannii ATCC 17978 at 28°C and 37°C. (A) The bacteria were able to cover the entire surface of the plate after 18 h of incubation at 37°C, but growth was significantly less at 28°C than at 37°C. Each data point represents a biological replicate. (B) The picture shows representative plates inoculated with A. baumannii ATCC 17978 and incubated at 28°C and 37°C. Statistical analysis was carried out using a paired t test.

    Techniques Used: Incubation

    40) Product Images from "Effect of Incubation Temperature on Antibiotic Resistance and Virulence Factors of Acinetobacter baumannii ATCC 17978"

    Article Title: Effect of Incubation Temperature on Antibiotic Resistance and Virulence Factors of Acinetobacter baumannii ATCC 17978

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01514-17

    Comparison of biofilm formation by A. baumannii ATCC 17978 at 28°C and 37°C. Biofilm formation was significantly greater at 28°C than at 37°C. Statistical analysis was carried out using two-way analysis of variance (ANOVA). *, P = 0.001.
    Figure Legend Snippet: Comparison of biofilm formation by A. baumannii ATCC 17978 at 28°C and 37°C. Biofilm formation was significantly greater at 28°C than at 37°C. Statistical analysis was carried out using two-way analysis of variance (ANOVA). *, P = 0.001.

    Techniques Used:

    Comparison of twitching motility of A. baumannii ATCC 17978 at 28°C and 37°C. (A) The bacteria were able to cover the entire surface of the plate after 18 h of incubation at 37°C, but growth was significantly less at 28°C than at 37°C. Each data point represents a biological replicate. (B) The picture shows representative plates inoculated with A. baumannii ATCC 17978 and incubated at 28°C and 37°C. Statistical analysis was carried out using a paired t test.
    Figure Legend Snippet: Comparison of twitching motility of A. baumannii ATCC 17978 at 28°C and 37°C. (A) The bacteria were able to cover the entire surface of the plate after 18 h of incubation at 37°C, but growth was significantly less at 28°C than at 37°C. Each data point represents a biological replicate. (B) The picture shows representative plates inoculated with A. baumannii ATCC 17978 and incubated at 28°C and 37°C. Statistical analysis was carried out using a paired t test.

    Techniques Used: Incubation

    Related Articles

    Produced:

    Article Title: Subinhibitory Concentrations of Trimethoprim and Sulfamethoxazole Prevent Biofilm Formation by Acinetobacter baumannii through Inhibition of Csu Pilus Expression
    Article Snippet: .. We found that, although ATCC 17978 was resistant to sulfamethoxazole (Smx) and trimethoprim (Tmp), subinhibitory concentrations of these antibiotics abolished the expression of Csu and consequently produced a dramatic reduction in biofilm formation by ATCC 17978. ..

    Mutagenesis:

    Article Title: Curcumin alleviates persistence of Acinetobacter baumannii against colistin
    Article Snippet: .. The efflux pump mutant strain A . baumannii ∆ adeB ATCC 17978 was obtained as gift from Maria Tomas from the Department of Microbiology, Complejo Hospitalario Universitario A Coruña-INIBIC, La Coruña, Spain . ..

    Expressing:

    Article Title: Subinhibitory Concentrations of Trimethoprim and Sulfamethoxazole Prevent Biofilm Formation by Acinetobacter baumannii through Inhibition of Csu Pilus Expression
    Article Snippet: .. We found that, although ATCC 17978 was resistant to sulfamethoxazole (Smx) and trimethoprim (Tmp), subinhibitory concentrations of these antibiotics abolished the expression of Csu and consequently produced a dramatic reduction in biofilm formation by ATCC 17978. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 91
    ATCC a baumannii strain atcc 17978
    ATCC 17978 mutants (A) and 29D2 mutants (B) deficient in surface-associated motility. Wildtypes and mutants of strains ATCC 17978 and 29D2 were inoculated on motility plates. Plates were incubated for 16 h at 37°C. The diameter (Ø) of the surface-associated motility spreading zone was measured and triplicates were statistically analyzed. All mutants of strains ATCC 17978 ( A ) and 29D2 ( B ) displayed a significant motility deficiency compared to their respective parental strain (p-value ≤ 0.05).
    A Baumannii Strain Atcc 17978, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a baumannii strain atcc 17978/product/ATCC
    Average 91 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    a baumannii strain atcc 17978 - by Bioz Stars, 2020-08
    91/100 stars
      Buy from Supplier

    91
    ATCC acinetobacter baumannii atcc 17978
    Antimicrobial resistance genes  (A)  and virulence genes  (B)  predicted in  A. baumannii  strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.
    Acinetobacter Baumannii Atcc 17978, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/acinetobacter baumannii atcc 17978/product/ATCC
    Average 91 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    acinetobacter baumannii atcc 17978 - by Bioz Stars, 2020-08
    91/100 stars
      Buy from Supplier

    93
    ATCC a baumannii atcc 17978ω tet
    (A) Appearance of antimicrobial-resistant A. <t>baumannii</t> <t>ATCC</t> 17978 derivatives following exposure to eDNA collected from the parent A. baumannii NU-60 strain. AMK, amikacin; CAR, carbenicillin; <t>TET,</t> tetracycline. (B) Number of CFU of antimicrobial-resistant derivatives obtained after exposure to eDNA collected from culture supernatants over 48 h. The eDNA collected from 20 ml of culture supernatant at each time point was used for one assay ( n = 3; means ± standard deviations [SD]). (C) Quantification of eDNA collected from 20 ml of culture supernatants (black bars) and optical density (O.D.) at 600 nm of bacterial cultures (black circles) ( n = 3; means ± SD). N.D., not detected.
    A Baumannii Atcc 17978ω Tet, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a baumannii atcc 17978ω tet/product/ATCC
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    a baumannii atcc 17978ω tet - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    93
    ATCC a baumannii atcc 17978
    PCR confirmation of mini-Tn 7 T-Apr-LAC and mini-Tn 7 T-Zeo-LAC genomic insertions in the A. <t>baumannii</t> strains used in this study. The mini-Tn 7 T-Apr-LAC and mini-Tn 7 T-Zeo-LAC genomic insertions in A. baumannii strains were confirmed using the ABglmS_F_New and Tn 7 R primers, which give the expected 368-bp band. Lane M, 100-bp molecular size marker (New England Biolabs); lane 1, A. baumannii ATCC 17978::mini-Tn 7 T-Apr-LAC; lane 2, A. baumannii ATCC 17978::mini-Tn 7 T-Zeo-LAC; lane 3, negative control for A. baumannii ATCC 17978; lane 4, A. baumannii AB030::mini-Tn 7 T-Apr-LAC; lane 5, negative control for A. baumannii AB030; lane 6, A. baumannii AB031::mini-Tn 7 T-Apr-LAC; lane 7, A. baumannii AB031::mini-Tn 7 T-Zeo-LAC; lane 8, negative control for A. baumannii AB031; lane 9, A. baumannii LAC-4::mini-Tn 7 T-Apr-LAC; lane 10, A. baumannii LAC-4::mini-Tn 7 T-Zeo-LAC; lane 11, negative control for A. baumannii LAC-4.
    A Baumannii Atcc 17978, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a baumannii atcc 17978/product/ATCC
    Average 93 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    a baumannii atcc 17978 - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    Image Search Results


    ATCC 17978 mutants (A) and 29D2 mutants (B) deficient in surface-associated motility. Wildtypes and mutants of strains ATCC 17978 and 29D2 were inoculated on motility plates. Plates were incubated for 16 h at 37°C. The diameter (Ø) of the surface-associated motility spreading zone was measured and triplicates were statistically analyzed. All mutants of strains ATCC 17978 ( A ) and 29D2 ( B ) displayed a significant motility deficiency compared to their respective parental strain (p-value ≤ 0.05).

    Journal: bioRxiv

    Article Title: Novel genes required for surface-associated motility in Acinetobacter baumannii

    doi: 10.1101/2020.03.18.992537

    Figure Lengend Snippet: ATCC 17978 mutants (A) and 29D2 mutants (B) deficient in surface-associated motility. Wildtypes and mutants of strains ATCC 17978 and 29D2 were inoculated on motility plates. Plates were incubated for 16 h at 37°C. The diameter (Ø) of the surface-associated motility spreading zone was measured and triplicates were statistically analyzed. All mutants of strains ATCC 17978 ( A ) and 29D2 ( B ) displayed a significant motility deficiency compared to their respective parental strain (p-value ≤ 0.05).

    Article Snippet: A recent study revealed the regulatory control of surface-associated motility and biofilm formation by a cyclic-di-GMP signaling network in A. baumannii strain ATCC 17978 [ ].

    Techniques: Incubation, Significance Assay

    Growth deficiency of mutant strains from ATCC 17978 (A) and 29D2 (B) mutant libraries. OD-adjusted bacterial cultures were grown for 9 h at 37°C under constant shaking. Every hour cultures were measured at an OD of 600 nm. For each strain data obtained from 3 independent cultures grown on the same day were averaged and represented by the mean ± SD. Growth defects compared to wildtype ATCC 17978 were observed for mutants that are involved in purine, pyrimidine, and diaminopropane biosynthesis ( A ). In strain 29D2, growth defects were observed for mutants involved in purine/pyrimidine/folate and diaminopropane biosynthesis, and for mutants  galE::Km ,  ompA::Km,  and  prpF::Km  ( B ). See Supplementary Figs. S4 and S5 for growth curves of all strains described in this study.

    Journal: bioRxiv

    Article Title: Novel genes required for surface-associated motility in Acinetobacter baumannii

    doi: 10.1101/2020.03.18.992537

    Figure Lengend Snippet: Growth deficiency of mutant strains from ATCC 17978 (A) and 29D2 (B) mutant libraries. OD-adjusted bacterial cultures were grown for 9 h at 37°C under constant shaking. Every hour cultures were measured at an OD of 600 nm. For each strain data obtained from 3 independent cultures grown on the same day were averaged and represented by the mean ± SD. Growth defects compared to wildtype ATCC 17978 were observed for mutants that are involved in purine, pyrimidine, and diaminopropane biosynthesis ( A ). In strain 29D2, growth defects were observed for mutants involved in purine/pyrimidine/folate and diaminopropane biosynthesis, and for mutants galE::Km , ompA::Km, and prpF::Km ( B ). See Supplementary Figs. S4 and S5 for growth curves of all strains described in this study.

    Article Snippet: A recent study revealed the regulatory control of surface-associated motility and biofilm formation by a cyclic-di-GMP signaling network in A. baumannii strain ATCC 17978 [ ].

    Techniques: Mutagenesis

    Pellicle biofilm formation of ATCC 17978 wildtype and mutants (A) and 29D2 wildtype and mutants (B). A. baumannii pellicle biofilms developed within 3 days of incubation and were stained with a 0.5% crystal violet solution. The biofilm was scrubbed and flushed off the tube walls and the absorption was determined at 550 nm. For each strain 3 independent experiments were performed and statistical significance was analyzed by the Student’s t test (2-tailed, unpaired). Significance as indicated: *, p-value ≤ 0.05; **, p-value ≤ 0.01; ***, p-value ≤ 0.001; ****, p-value ≤ 0.0001.

    Journal: bioRxiv

    Article Title: Novel genes required for surface-associated motility in Acinetobacter baumannii

    doi: 10.1101/2020.03.18.992537

    Figure Lengend Snippet: Pellicle biofilm formation of ATCC 17978 wildtype and mutants (A) and 29D2 wildtype and mutants (B). A. baumannii pellicle biofilms developed within 3 days of incubation and were stained with a 0.5% crystal violet solution. The biofilm was scrubbed and flushed off the tube walls and the absorption was determined at 550 nm. For each strain 3 independent experiments were performed and statistical significance was analyzed by the Student’s t test (2-tailed, unpaired). Significance as indicated: *, p-value ≤ 0.05; **, p-value ≤ 0.01; ***, p-value ≤ 0.001; ****, p-value ≤ 0.0001.

    Article Snippet: A recent study revealed the regulatory control of surface-associated motility and biofilm formation by a cyclic-di-GMP signaling network in A. baumannii strain ATCC 17978 [ ].

    Techniques: Incubation, Staining

    Genes inactivated in A. baumannii ATCC 17978 mutants with a surface-associated motility defect and their known/predicted/putative function in the bacterial cell. A common color code indicates that mutants belong to the same functional, processual, and/or structural category. OM, outer membrane; IM, inner membrane; Ap4A, diadenosine tetraphosphate; Ap5A, diadenosine pentaphosphate; SAM, S-adenosyl-L-methionine; SAH, S-adenosylhomocysteine; Ado, adenosine; Hcy, homocysteine; Met, methionine; ASA, L-aspartate 4-semialdehyde; DAB, L-2,4-diaminobutanoate; 2-MMC, 2-methylcitric acid cycle.

    Journal: bioRxiv

    Article Title: Novel genes required for surface-associated motility in Acinetobacter baumannii

    doi: 10.1101/2020.03.18.992537

    Figure Lengend Snippet: Genes inactivated in A. baumannii ATCC 17978 mutants with a surface-associated motility defect and their known/predicted/putative function in the bacterial cell. A common color code indicates that mutants belong to the same functional, processual, and/or structural category. OM, outer membrane; IM, inner membrane; Ap4A, diadenosine tetraphosphate; Ap5A, diadenosine pentaphosphate; SAM, S-adenosyl-L-methionine; SAH, S-adenosylhomocysteine; Ado, adenosine; Hcy, homocysteine; Met, methionine; ASA, L-aspartate 4-semialdehyde; DAB, L-2,4-diaminobutanoate; 2-MMC, 2-methylcitric acid cycle.

    Article Snippet: A recent study revealed the regulatory control of surface-associated motility and biofilm formation by a cyclic-di-GMP signaling network in A. baumannii strain ATCC 17978 [ ].

    Techniques: Functional Assay

    Attenuation of A. baumannii ATCC 17978 mutants (A) and 29D2 mutants (B) in the G. mellonella caterpillar infection model. Caterpillars were infected with 3 x 10 5 CFU of A. baumannii strains as indicated. Sterile PBS (black lines) was used as a control. Three independent experiments were performed with groups of 16 caterpillars for every bacterial strain and control. Data obtained from 3 independent experiments were averaged and represented by the mean ± SD. In strain ATCC 17978, 15 of 30 mutants showed a significant attenuation at 5 days post-infection (see Table 1 for p-values) compared to the wildtype strain ( A ). In strain 29D2, 11 of 21 mutants were attenuated (see Table 2 for p-values) in the G. mellonella infection model ( B ). See Supplementary Figs. S6 and S7 for infection data of all strains described in this study.

    Journal: bioRxiv

    Article Title: Novel genes required for surface-associated motility in Acinetobacter baumannii

    doi: 10.1101/2020.03.18.992537

    Figure Lengend Snippet: Attenuation of A. baumannii ATCC 17978 mutants (A) and 29D2 mutants (B) in the G. mellonella caterpillar infection model. Caterpillars were infected with 3 x 10 5 CFU of A. baumannii strains as indicated. Sterile PBS (black lines) was used as a control. Three independent experiments were performed with groups of 16 caterpillars for every bacterial strain and control. Data obtained from 3 independent experiments were averaged and represented by the mean ± SD. In strain ATCC 17978, 15 of 30 mutants showed a significant attenuation at 5 days post-infection (see Table 1 for p-values) compared to the wildtype strain ( A ). In strain 29D2, 11 of 21 mutants were attenuated (see Table 2 for p-values) in the G. mellonella infection model ( B ). See Supplementary Figs. S6 and S7 for infection data of all strains described in this study.

    Article Snippet: A recent study revealed the regulatory control of surface-associated motility and biofilm formation by a cyclic-di-GMP signaling network in A. baumannii strain ATCC 17978 [ ].

    Techniques: Infection

    Antimicrobial resistance genes  (A)  and virulence genes  (B)  predicted in  A. baumannii  strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.

    Journal: International journal of medical microbiology : IJMM

    Article Title: Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606

    doi: 10.1016/j.ijmm.2020.151412

    Figure Lengend Snippet: Antimicrobial resistance genes (A) and virulence genes (B) predicted in A. baumannii strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.

    Article Snippet: GI sul2 , a genomic island carrying the sul2 sulphonamide resistance gene and the small mobile element CR2 found in the Enterobacter cloacae subspecies cloacae type strain ATCC 13047 from 1890, Shigella flexneri ATCC 700930 from 1954 and Acinetobacter baumannii ATCC 17978 from 1951 .

    Techniques:

    (A) Appearance of antimicrobial-resistant A. baumannii ATCC 17978 derivatives following exposure to eDNA collected from the parent A. baumannii NU-60 strain. AMK, amikacin; CAR, carbenicillin; TET, tetracycline. (B) Number of CFU of antimicrobial-resistant derivatives obtained after exposure to eDNA collected from culture supernatants over 48 h. The eDNA collected from 20 ml of culture supernatant at each time point was used for one assay ( n = 3; means ± standard deviations [SD]). (C) Quantification of eDNA collected from 20 ml of culture supernatants (black bars) and optical density (O.D.) at 600 nm of bacterial cultures (black circles) ( n = 3; means ± SD). N.D., not detected.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Intercellular Transfer of Chromosomal Antimicrobial Resistance Genes between Acinetobacter baumannii Strains Mediated by Prophages

    doi: 10.1128/AAC.00334-19

    Figure Lengend Snippet: (A) Appearance of antimicrobial-resistant A. baumannii ATCC 17978 derivatives following exposure to eDNA collected from the parent A. baumannii NU-60 strain. AMK, amikacin; CAR, carbenicillin; TET, tetracycline. (B) Number of CFU of antimicrobial-resistant derivatives obtained after exposure to eDNA collected from culture supernatants over 48 h. The eDNA collected from 20 ml of culture supernatant at each time point was used for one assay ( n = 3; means ± standard deviations [SD]). (C) Quantification of eDNA collected from 20 ml of culture supernatants (black bars) and optical density (O.D.) at 600 nm of bacterial cultures (black circles) ( n = 3; means ± SD). N.D., not detected.

    Article Snippet: The transfer of tet (B) was confirmed by nucleotide sequencing of A. baumannii ATCC 17978Ω tet (B) (Fig. S3).

    Techniques:

    PCR confirmation of mini-Tn 7 T-Apr-LAC and mini-Tn 7 T-Zeo-LAC genomic insertions in the A. baumannii strains used in this study. The mini-Tn 7 T-Apr-LAC and mini-Tn 7 T-Zeo-LAC genomic insertions in A. baumannii strains were confirmed using the ABglmS_F_New and Tn 7 R primers, which give the expected 368-bp band. Lane M, 100-bp molecular size marker (New England Biolabs); lane 1, A. baumannii ATCC 17978::mini-Tn 7 T-Apr-LAC; lane 2, A. baumannii ATCC 17978::mini-Tn 7 T-Zeo-LAC; lane 3, negative control for A. baumannii ATCC 17978; lane 4, A. baumannii AB030::mini-Tn 7 T-Apr-LAC; lane 5, negative control for A. baumannii AB030; lane 6, A. baumannii AB031::mini-Tn 7 T-Apr-LAC; lane 7, A. baumannii AB031::mini-Tn 7 T-Zeo-LAC; lane 8, negative control for A. baumannii AB031; lane 9, A. baumannii LAC-4::mini-Tn 7 T-Apr-LAC; lane 10, A. baumannii LAC-4::mini-Tn 7 T-Zeo-LAC; lane 11, negative control for A. baumannii LAC-4.

    Journal: Applied and Environmental Microbiology

    Article Title: Next Generation of Tn7-Based Single-Copy Insertion Elements for Use in Multi- and Pan-Drug-Resistant Strains of Acinetobacter baumannii

    doi: 10.1128/AEM.00066-19

    Figure Lengend Snippet: PCR confirmation of mini-Tn 7 T-Apr-LAC and mini-Tn 7 T-Zeo-LAC genomic insertions in the A. baumannii strains used in this study. The mini-Tn 7 T-Apr-LAC and mini-Tn 7 T-Zeo-LAC genomic insertions in A. baumannii strains were confirmed using the ABglmS_F_New and Tn 7 R primers, which give the expected 368-bp band. Lane M, 100-bp molecular size marker (New England Biolabs); lane 1, A. baumannii ATCC 17978::mini-Tn 7 T-Apr-LAC; lane 2, A. baumannii ATCC 17978::mini-Tn 7 T-Zeo-LAC; lane 3, negative control for A. baumannii ATCC 17978; lane 4, A. baumannii AB030::mini-Tn 7 T-Apr-LAC; lane 5, negative control for A. baumannii AB030; lane 6, A. baumannii AB031::mini-Tn 7 T-Apr-LAC; lane 7, A. baumannii AB031::mini-Tn 7 T-Zeo-LAC; lane 8, negative control for A. baumannii AB031; lane 9, A. baumannii LAC-4::mini-Tn 7 T-Apr-LAC; lane 10, A. baumannii LAC-4::mini-Tn 7 T-Zeo-LAC; lane 11, negative control for A. baumannii LAC-4.

    Article Snippet: The genome sequences of A. baumannii ATCC 17978 , AB030 , AB031 , and LAC-4 ( ) were used to determine the presence of att Tn 7 ).

    Techniques: Polymerase Chain Reaction, Marker, Negative Control

    Deletion of apramycin or zeocin resistance markers from A. baumannii using pFLP2A. (a) Photographs of the LB agar plate (left) and the LB agar plate supplemented with 100 µg/ml of apramycin (right) to screen for the loss of the Apr r resistance gene by using pFLP2Z, encoding Flp recombinase. The absence of growth on LB plus Apr confirms the loss of aac(3)-IV (Apr r marker). (b) Photographs of the LB agar plate (left) and the LB agar plate supplemented with 100 µg/ml of zeocin (right) to screen for the loss of Zeo r resistance gene. The absence of growth on LB plus Zeo confirms the loss of ble (Zeo r marker). (c) The loss of the relevant resistance markers was further confirmed with PCR. Lanes 1, 3, and 5 show the 998-bp band corresponding to the aac(3)-IV gene in ATCC 17978, LAC-4, and AB031, respectively. Lanes 2, 4, and 6 show the loss of the 998-bp band in the pFLP2Z-treated ATCC 17978, LAC-4, and AB031 strains, respectively. Lanes 7 and 8, positive and negative controls for the 998-bp aac(3)-IV band, respectively. Lanes 9, 11, and 13 show the 569-bp band corresponding to the ble gene in ATCC 17978, LAC-4, and AB031, respectively. Lanes 10, 12, and 14 show the loss of the 569-bp band in the pFLP2A-treated ATCC 17978, LAC-4, and AB031 strains, respectively. Lanes 15 and 16, positive and negative controls for the 569-bp ble band, respectively. Lanes M, 100-bp molecular size marker.

    Journal: Applied and Environmental Microbiology

    Article Title: Next Generation of Tn7-Based Single-Copy Insertion Elements for Use in Multi- and Pan-Drug-Resistant Strains of Acinetobacter baumannii

    doi: 10.1128/AEM.00066-19

    Figure Lengend Snippet: Deletion of apramycin or zeocin resistance markers from A. baumannii using pFLP2A. (a) Photographs of the LB agar plate (left) and the LB agar plate supplemented with 100 µg/ml of apramycin (right) to screen for the loss of the Apr r resistance gene by using pFLP2Z, encoding Flp recombinase. The absence of growth on LB plus Apr confirms the loss of aac(3)-IV (Apr r marker). (b) Photographs of the LB agar plate (left) and the LB agar plate supplemented with 100 µg/ml of zeocin (right) to screen for the loss of Zeo r resistance gene. The absence of growth on LB plus Zeo confirms the loss of ble (Zeo r marker). (c) The loss of the relevant resistance markers was further confirmed with PCR. Lanes 1, 3, and 5 show the 998-bp band corresponding to the aac(3)-IV gene in ATCC 17978, LAC-4, and AB031, respectively. Lanes 2, 4, and 6 show the loss of the 998-bp band in the pFLP2Z-treated ATCC 17978, LAC-4, and AB031 strains, respectively. Lanes 7 and 8, positive and negative controls for the 998-bp aac(3)-IV band, respectively. Lanes 9, 11, and 13 show the 569-bp band corresponding to the ble gene in ATCC 17978, LAC-4, and AB031, respectively. Lanes 10, 12, and 14 show the loss of the 569-bp band in the pFLP2A-treated ATCC 17978, LAC-4, and AB031 strains, respectively. Lanes 15 and 16, positive and negative controls for the 569-bp ble band, respectively. Lanes M, 100-bp molecular size marker.

    Article Snippet: The genome sequences of A. baumannii ATCC 17978 , AB030 , AB031 , and LAC-4 ( ) were used to determine the presence of att Tn 7 ).

    Techniques: Marker, Polymerase Chain Reaction

    Sequence alignment of the att Tn 7 -associated sequence and the Tn 7 insertion site of Acinetobacter baumannii ATCC 17978 and clinical isolates used in this study, showing the highly conserved sequence homology between the strains. The insertion site is located 24 bp downstream of the glmS2 gene in a neutral location (as shown by the arrow). The stop codon of the glmS2 gene is outlined in the red box. The consensus sequence is shown at the bottom of the alignment and displays 100% identity.

    Journal: Applied and Environmental Microbiology

    Article Title: Next Generation of Tn7-Based Single-Copy Insertion Elements for Use in Multi- and Pan-Drug-Resistant Strains of Acinetobacter baumannii

    doi: 10.1128/AEM.00066-19

    Figure Lengend Snippet: Sequence alignment of the att Tn 7 -associated sequence and the Tn 7 insertion site of Acinetobacter baumannii ATCC 17978 and clinical isolates used in this study, showing the highly conserved sequence homology between the strains. The insertion site is located 24 bp downstream of the glmS2 gene in a neutral location (as shown by the arrow). The stop codon of the glmS2 gene is outlined in the red box. The consensus sequence is shown at the bottom of the alignment and displays 100% identity.

    Article Snippet: The genome sequences of A. baumannii ATCC 17978 , AB030 , AB031 , and LAC-4 ( ) were used to determine the presence of att Tn 7 ).

    Techniques: Sequencing