inactive ppad  (ATCC)


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  • 92
    Name:
    Treponema denticola a CIP 103919 DSM 14222
    Description:

    Catalog Number:
    35405
    Price:
    None
    Applications:
    Produces major surface proteinProduces methyl-accepting chemotaxis proteinProduces prolyl aminopeptidase proline iminopeptidase
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    Structured Review

    ATCC inactive ppad
    Identification and functional annotation of TIGK genes significantly dependent on <t>PPAD</t> activity. ( A ) Scatterplot of log2 fold-change (log2FC) in gene expression of TIGKs infected with P. gingivalis WT vs. control (y-axis), and P. gingivalis PPAD <t>C351A</t> vs. P. gingivalis WT (x-axis). Genes altered exclusively in the P. gingivalis WT vs. control comparison are displayed in red when significantly up-regulated (log2FC > 0.5, FDR q

    https://www.bioz.com/result/inactive ppad/product/ATCC
    Average 92 stars, based on 353 article reviews
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    inactive ppad - by Bioz Stars, 2020-08
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    Images

    1) Product Images from "Impact of Porphyromonas gingivalis Peptidylarginine Deiminase on Bacterial Biofilm Formation, Epithelial Cell Invasion, and Epithelial Cell Transcriptional Landscape"

    Article Title: Impact of Porphyromonas gingivalis Peptidylarginine Deiminase on Bacterial Biofilm Formation, Epithelial Cell Invasion, and Epithelial Cell Transcriptional Landscape

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-32603-y

    Identification and functional annotation of TIGK genes significantly dependent on PPAD activity. ( A ) Scatterplot of log2 fold-change (log2FC) in gene expression of TIGKs infected with P. gingivalis WT vs. control (y-axis), and P. gingivalis PPAD C351A vs. P. gingivalis WT (x-axis). Genes altered exclusively in the P. gingivalis WT vs. control comparison are displayed in red when significantly up-regulated (log2FC > 0.5, FDR q
    Figure Legend Snippet: Identification and functional annotation of TIGK genes significantly dependent on PPAD activity. ( A ) Scatterplot of log2 fold-change (log2FC) in gene expression of TIGKs infected with P. gingivalis WT vs. control (y-axis), and P. gingivalis PPAD C351A vs. P. gingivalis WT (x-axis). Genes altered exclusively in the P. gingivalis WT vs. control comparison are displayed in red when significantly up-regulated (log2FC > 0.5, FDR q

    Techniques Used: Functional Assay, Activity Assay, Expressing, Infection

    The effect of P. gingivalis PPAD on bacterial abundance ( a ), species composition ( b ) and ( c ) the biofilm structure in multispecies biofilms. A biofilm consisting of T. forsythia , F. nucleatum , A. naeslundii , S. gordonii , and one of three P. gingivalis strains, wild type (WT), ppad deletion strain ( Δppad ), or a strain harboring inactivated PPAD (C351A), was cultured for 48 h, following which ( a,b ) the bacterial DNA was extracted and assessed via qPCR, ( c ) biofilm was fixed and observed via SEM under 1500x magnification. The ( a ) bacterial load and ( b ) species composition of each biofilm model are presented as mean ± SD from three independent experiments. Data were plotted on a logarithmic scale.
    Figure Legend Snippet: The effect of P. gingivalis PPAD on bacterial abundance ( a ), species composition ( b ) and ( c ) the biofilm structure in multispecies biofilms. A biofilm consisting of T. forsythia , F. nucleatum , A. naeslundii , S. gordonii , and one of three P. gingivalis strains, wild type (WT), ppad deletion strain ( Δppad ), or a strain harboring inactivated PPAD (C351A), was cultured for 48 h, following which ( a,b ) the bacterial DNA was extracted and assessed via qPCR, ( c ) biofilm was fixed and observed via SEM under 1500x magnification. The ( a ) bacterial load and ( b ) species composition of each biofilm model are presented as mean ± SD from three independent experiments. Data were plotted on a logarithmic scale.

    Techniques Used: Cell Culture, Real-time Polymerase Chain Reaction

    2) Product Images from "Genome-Wide Relatedness of Treponema pedis, from Gingiva and Necrotic Skin Lesions of Pigs, with the Human Oral Pathogen Treponema denticola"

    Article Title: Genome-Wide Relatedness of Treponema pedis, from Gingiva and Necrotic Skin Lesions of Pigs, with the Human Oral Pathogen Treponema denticola

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0071281

    Circular representation of the T. pedis T A4 genome and complete genome alignment with T. denticola . (A.) Circular representation of the T. pedis T A4 genome. The CDSs are shown in violet where the outer circle represents predictions on the plus strand and the second circle those on the minus strand. CDSs with a best BLASTP hit in T. denticola ATCC 35405 are colored red and shown in the third circle. The fourth circle represents genes with best BLASTP hits in T. brennaborense (black), F. nucleatum (green), F. alocis (blue) and T. succinifaciens (grey). G+C skew is drawn in the inner circle. (B.) Complete genome alignment between T. pedis T A4 and T. denticola ATCC 35405. Dots represent maximum unique matches (MUMs) between the genomes. MUMs oriented in the same direction are depicted as red dots and reverse complemented MUMs are depicted as blue dots.
    Figure Legend Snippet: Circular representation of the T. pedis T A4 genome and complete genome alignment with T. denticola . (A.) Circular representation of the T. pedis T A4 genome. The CDSs are shown in violet where the outer circle represents predictions on the plus strand and the second circle those on the minus strand. CDSs with a best BLASTP hit in T. denticola ATCC 35405 are colored red and shown in the third circle. The fourth circle represents genes with best BLASTP hits in T. brennaborense (black), F. nucleatum (green), F. alocis (blue) and T. succinifaciens (grey). G+C skew is drawn in the inner circle. (B.) Complete genome alignment between T. pedis T A4 and T. denticola ATCC 35405. Dots represent maximum unique matches (MUMs) between the genomes. MUMs oriented in the same direction are depicted as red dots and reverse complemented MUMs are depicted as blue dots.

    Techniques Used:

    3) Product Images from "Anti-HmuY Antibodies Specifically Recognize Porphyromonas gingivalis HmuY Protein but Not Homologous Proteins in Other Periodontopathogens"

    Article Title: Anti-HmuY Antibodies Specifically Recognize Porphyromonas gingivalis HmuY Protein but Not Homologous Proteins in Other Periodontopathogens

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0117508

    Reactivity of anti-HmuY antibodies with bacteria. ELISA was carried out using antibodies raised against purified HmuY protein (anti-HmuY 1) and P . gingivalis wild-type (A7436 and ATCC 33277) and hmuY mutant (TO4) strains. Bacteria were cultured in high iron/heme (A) or low-iron/heme (B) media. No reactivity was observed for anti-HmuY 1–1 (epitope 1), anti-HmuY 1–2 (epitope 2), or anti-HmuY 1–3 (epitope 3) antibodies. Live bacteria were used as antigens.
    Figure Legend Snippet: Reactivity of anti-HmuY antibodies with bacteria. ELISA was carried out using antibodies raised against purified HmuY protein (anti-HmuY 1) and P . gingivalis wild-type (A7436 and ATCC 33277) and hmuY mutant (TO4) strains. Bacteria were cultured in high iron/heme (A) or low-iron/heme (B) media. No reactivity was observed for anti-HmuY 1–1 (epitope 1), anti-HmuY 1–2 (epitope 2), or anti-HmuY 1–3 (epitope 3) antibodies. Live bacteria were used as antigens.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Purification, Mutagenesis, Cell Culture

    Reactivity of anti-HmuY antibodies with P . gingivalis wild-type (A7436 and ATCC 33277) and hmuY mutant (TO4) strains. Bacteria were cultured in high iron/heme (Hm) or low-iron/heme (DIP) media. Western blotting was carried out using antibodies raised against purified HmuY protein (anti-HmuY 1), epitope 1 (anti-HmuY 1–1), epitope 2 (anti-HmuY 1–2), and epitope 3 (anti-HmuY 1–3). Arrows denote location of bands corresponding to P . gingivalis HmuY protein.
    Figure Legend Snippet: Reactivity of anti-HmuY antibodies with P . gingivalis wild-type (A7436 and ATCC 33277) and hmuY mutant (TO4) strains. Bacteria were cultured in high iron/heme (Hm) or low-iron/heme (DIP) media. Western blotting was carried out using antibodies raised against purified HmuY protein (anti-HmuY 1), epitope 1 (anti-HmuY 1–1), epitope 2 (anti-HmuY 1–2), and epitope 3 (anti-HmuY 1–3). Arrows denote location of bands corresponding to P . gingivalis HmuY protein.

    Techniques Used: Mutagenesis, Cell Culture, Western Blot, Purification

    4) Product Images from "Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms"

    Article Title: Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

    Journal: Molecular Oral Microbiology

    doi: 10.1111/omi.12182

    Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)
    Figure Legend Snippet: Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)

    Techniques Used: Mutagenesis

    5) Product Images from "Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms"

    Article Title: Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

    Journal: Molecular Oral Microbiology

    doi: 10.1111/omi.12182

    Dual fluorescence in situ hybridization staining of Tannerella forsythia and Campylobacter rectus for biofilms harboring ATCC 43037 wild‐type (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: T. forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)
    Figure Legend Snippet: Dual fluorescence in situ hybridization staining of Tannerella forsythia and Campylobacter rectus for biofilms harboring ATCC 43037 wild‐type (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: T. forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)

    Techniques Used: Fluorescence, In Situ Hybridization, Staining

    Fluorescence in situ hybridization staining of biofilms harboring Tannerella forsythia ATCC 43037 mutants (A) ∆pseC , (B) ∆wecC , and (C) ∆tfs AB . Red: T. forsythia , cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A) and 10 μm (B, C)
    Figure Legend Snippet: Fluorescence in situ hybridization staining of biofilms harboring Tannerella forsythia ATCC 43037 mutants (A) ∆pseC , (B) ∆wecC , and (C) ∆tfs AB . Red: T. forsythia , cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A) and 10 μm (B, C)

    Techniques Used: Fluorescence, In Situ Hybridization, Staining

    Comparison of colony‐forming unit ( CFU ) counting and quantitative polymerase chain reaction ( qPCR ) for Tannerella forsythia wild‐type strains and mutants in the subgingival “Zurich biofilm”. Total bacteria for 10‐species biofilms with different T. forsythia strains and mutants enumerated by CFU counts (red boxes) and qPCR (blue boxes) for three independent experiments with three technical replicates, each, are shown (Whiskers boxplots 5th to 95th centile)
    Figure Legend Snippet: Comparison of colony‐forming unit ( CFU ) counting and quantitative polymerase chain reaction ( qPCR ) for Tannerella forsythia wild‐type strains and mutants in the subgingival “Zurich biofilm”. Total bacteria for 10‐species biofilms with different T. forsythia strains and mutants enumerated by CFU counts (red boxes) and qPCR (blue boxes) for three independent experiments with three technical replicates, each, are shown (Whiskers boxplots 5th to 95th centile)

    Techniques Used: Real-time Polymerase Chain Reaction

    Box plots showing cell numbers of all species determined by quantitative real‐time PCR for biofilms with Tannerella forsythia ATCC 43037 wild‐type or mutants (∆pseC , ∆wecC , ∆tfs AB , ∆pseC comp ) (A) and UB 4 wild‐type or mutants (∆legC , ∆legC comp ), respectively (B). Data derived from three independent experiments were plotted on a logarithmic scale. Asterisk (*) indicates significant differences ( P ≤.05) between the groups
    Figure Legend Snippet: Box plots showing cell numbers of all species determined by quantitative real‐time PCR for biofilms with Tannerella forsythia ATCC 43037 wild‐type or mutants (∆pseC , ∆wecC , ∆tfs AB , ∆pseC comp ) (A) and UB 4 wild‐type or mutants (∆legC , ∆legC comp ), respectively (B). Data derived from three independent experiments were plotted on a logarithmic scale. Asterisk (*) indicates significant differences ( P ≤.05) between the groups

    Techniques Used: Real-time Polymerase Chain Reaction, Derivative Assay

    Comparison of 10‐species biofilms with two Tannerella forsythia wild‐type strains. (A) Whiskers boxplots (5th to 95th centile) show bacterial numbers determined by quantitative real‐time PCR from three independent experiments. Asterisk (*) indicates a statistically significant difference ( P ≤.05) between groups. The two groups represent biofilms with either T. forsythia ATCC 43037 wild‐type or T. forsythia UB 4 wild‐type. (B, C) Fluorescence in situ hybridization stainings of fixed biofilms showing the localization of ATCC 43037 wild‐type (B) and UB 4 wild‐type (C). Red/yellow: T. forsythia; cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Here a representative area for one disk each is shown with a top view in the left panel and a side view with the biofilm–disk interface directed towards the top view; scale bars 5 μm (B) and 10 μm (C)
    Figure Legend Snippet: Comparison of 10‐species biofilms with two Tannerella forsythia wild‐type strains. (A) Whiskers boxplots (5th to 95th centile) show bacterial numbers determined by quantitative real‐time PCR from three independent experiments. Asterisk (*) indicates a statistically significant difference ( P ≤.05) between groups. The two groups represent biofilms with either T. forsythia ATCC 43037 wild‐type or T. forsythia UB 4 wild‐type. (B, C) Fluorescence in situ hybridization stainings of fixed biofilms showing the localization of ATCC 43037 wild‐type (B) and UB 4 wild‐type (C). Red/yellow: T. forsythia; cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Here a representative area for one disk each is shown with a top view in the left panel and a side view with the biofilm–disk interface directed towards the top view; scale bars 5 μm (B) and 10 μm (C)

    Techniques Used: Real-time Polymerase Chain Reaction, Fluorescence, In Situ Hybridization, Staining

    Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)
    Figure Legend Snippet: Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)

    Techniques Used: Mutagenesis

    6) Product Images from "Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms"

    Article Title: Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

    Journal: Molecular Oral Microbiology

    doi: 10.1111/omi.12182

    Dual fluorescence in situ hybridization staining of Tannerella forsythia and Campylobacter rectus for biofilms harboring ATCC 43037 wild‐type (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: T. forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)
    Figure Legend Snippet: Dual fluorescence in situ hybridization staining of Tannerella forsythia and Campylobacter rectus for biofilms harboring ATCC 43037 wild‐type (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: T. forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)

    Techniques Used: Fluorescence, In Situ Hybridization, Staining

    Fluorescence in situ hybridization staining of biofilms harboring Tannerella forsythia ATCC 43037 mutants (A) ∆pseC , (B) ∆wecC , and (C) ∆tfs AB . Red: T. forsythia , cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A) and 10 μm (B, C)
    Figure Legend Snippet: Fluorescence in situ hybridization staining of biofilms harboring Tannerella forsythia ATCC 43037 mutants (A) ∆pseC , (B) ∆wecC , and (C) ∆tfs AB . Red: T. forsythia , cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A) and 10 μm (B, C)

    Techniques Used: Fluorescence, In Situ Hybridization, Staining

    Comparison of colony‐forming unit ( CFU ) counting and quantitative polymerase chain reaction ( qPCR ) for Tannerella forsythia wild‐type strains and mutants in the subgingival “Zurich biofilm”. Total bacteria for 10‐species biofilms with different T. forsythia strains and mutants enumerated by CFU counts (red boxes) and qPCR (blue boxes) for three independent experiments with three technical replicates, each, are shown (Whiskers boxplots 5th to 95th centile)
    Figure Legend Snippet: Comparison of colony‐forming unit ( CFU ) counting and quantitative polymerase chain reaction ( qPCR ) for Tannerella forsythia wild‐type strains and mutants in the subgingival “Zurich biofilm”. Total bacteria for 10‐species biofilms with different T. forsythia strains and mutants enumerated by CFU counts (red boxes) and qPCR (blue boxes) for three independent experiments with three technical replicates, each, are shown (Whiskers boxplots 5th to 95th centile)

    Techniques Used: Real-time Polymerase Chain Reaction

    Box plots showing cell numbers of all species determined by quantitative real‐time PCR for biofilms with Tannerella forsythia ATCC 43037 wild‐type or mutants (∆pseC , ∆wecC , ∆tfs AB , ∆pseC comp ) (A) and UB 4 wild‐type or mutants (∆legC , ∆legC comp ), respectively (B). Data derived from three independent experiments were plotted on a logarithmic scale. Asterisk (*) indicates significant differences ( P ≤.05) between the groups
    Figure Legend Snippet: Box plots showing cell numbers of all species determined by quantitative real‐time PCR for biofilms with Tannerella forsythia ATCC 43037 wild‐type or mutants (∆pseC , ∆wecC , ∆tfs AB , ∆pseC comp ) (A) and UB 4 wild‐type or mutants (∆legC , ∆legC comp ), respectively (B). Data derived from three independent experiments were plotted on a logarithmic scale. Asterisk (*) indicates significant differences ( P ≤.05) between the groups

    Techniques Used: Real-time Polymerase Chain Reaction, Derivative Assay

    Comparison of 10‐species biofilms with two Tannerella forsythia wild‐type strains. (A) Whiskers boxplots (5th to 95th centile) show bacterial numbers determined by quantitative real‐time PCR from three independent experiments. Asterisk (*) indicates a statistically significant difference ( P ≤.05) between groups. The two groups represent biofilms with either T. forsythia ATCC 43037 wild‐type or T. forsythia UB 4 wild‐type. (B, C) Fluorescence in situ hybridization stainings of fixed biofilms showing the localization of ATCC 43037 wild‐type (B) and UB 4 wild‐type (C). Red/yellow: T. forsythia; cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Here a representative area for one disk each is shown with a top view in the left panel and a side view with the biofilm–disk interface directed towards the top view; scale bars 5 μm (B) and 10 μm (C)
    Figure Legend Snippet: Comparison of 10‐species biofilms with two Tannerella forsythia wild‐type strains. (A) Whiskers boxplots (5th to 95th centile) show bacterial numbers determined by quantitative real‐time PCR from three independent experiments. Asterisk (*) indicates a statistically significant difference ( P ≤.05) between groups. The two groups represent biofilms with either T. forsythia ATCC 43037 wild‐type or T. forsythia UB 4 wild‐type. (B, C) Fluorescence in situ hybridization stainings of fixed biofilms showing the localization of ATCC 43037 wild‐type (B) and UB 4 wild‐type (C). Red/yellow: T. forsythia; cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Here a representative area for one disk each is shown with a top view in the left panel and a side view with the biofilm–disk interface directed towards the top view; scale bars 5 μm (B) and 10 μm (C)

    Techniques Used: Real-time Polymerase Chain Reaction, Fluorescence, In Situ Hybridization, Staining

    Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)
    Figure Legend Snippet: Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)

    Techniques Used: Mutagenesis

    7) Product Images from "Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms"

    Article Title: Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

    Journal: Molecular Oral Microbiology

    doi: 10.1111/omi.12182

    Dual fluorescence in situ hybridization staining of Tannerella forsythia and Campylobacter rectus for biofilms harboring ATCC 43037 wild‐type (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: T. forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)
    Figure Legend Snippet: Dual fluorescence in situ hybridization staining of Tannerella forsythia and Campylobacter rectus for biofilms harboring ATCC 43037 wild‐type (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: T. forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)

    Techniques Used: Fluorescence, In Situ Hybridization, Staining

    Fluorescence in situ hybridization staining of biofilms harboring Tannerella forsythia ATCC 43037 mutants (A) ∆pseC , (B) ∆wecC , and (C) ∆tfs AB . Red: T. forsythia , cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A) and 10 μm (B, C)
    Figure Legend Snippet: Fluorescence in situ hybridization staining of biofilms harboring Tannerella forsythia ATCC 43037 mutants (A) ∆pseC , (B) ∆wecC , and (C) ∆tfs AB . Red: T. forsythia , cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A) and 10 μm (B, C)

    Techniques Used: Fluorescence, In Situ Hybridization, Staining

    Comparison of colony‐forming unit ( CFU ) counting and quantitative polymerase chain reaction ( qPCR ) for Tannerella forsythia wild‐type strains and mutants in the subgingival “Zurich biofilm”. Total bacteria for 10‐species biofilms with different T. forsythia strains and mutants enumerated by CFU counts (red boxes) and qPCR (blue boxes) for three independent experiments with three technical replicates, each, are shown (Whiskers boxplots 5th to 95th centile)
    Figure Legend Snippet: Comparison of colony‐forming unit ( CFU ) counting and quantitative polymerase chain reaction ( qPCR ) for Tannerella forsythia wild‐type strains and mutants in the subgingival “Zurich biofilm”. Total bacteria for 10‐species biofilms with different T. forsythia strains and mutants enumerated by CFU counts (red boxes) and qPCR (blue boxes) for three independent experiments with three technical replicates, each, are shown (Whiskers boxplots 5th to 95th centile)

    Techniques Used: Real-time Polymerase Chain Reaction

    Box plots showing cell numbers of all species determined by quantitative real‐time PCR for biofilms with Tannerella forsythia ATCC 43037 wild‐type or mutants (∆pseC , ∆wecC , ∆tfs AB , ∆pseC comp ) (A) and UB 4 wild‐type or mutants (∆legC , ∆legC comp ), respectively (B). Data derived from three independent experiments were plotted on a logarithmic scale. Asterisk (*) indicates significant differences ( P ≤.05) between the groups
    Figure Legend Snippet: Box plots showing cell numbers of all species determined by quantitative real‐time PCR for biofilms with Tannerella forsythia ATCC 43037 wild‐type or mutants (∆pseC , ∆wecC , ∆tfs AB , ∆pseC comp ) (A) and UB 4 wild‐type or mutants (∆legC , ∆legC comp ), respectively (B). Data derived from three independent experiments were plotted on a logarithmic scale. Asterisk (*) indicates significant differences ( P ≤.05) between the groups

    Techniques Used: Real-time Polymerase Chain Reaction, Derivative Assay

    Comparison of 10‐species biofilms with two Tannerella forsythia wild‐type strains. (A) Whiskers boxplots (5th to 95th centile) show bacterial numbers determined by quantitative real‐time PCR from three independent experiments. Asterisk (*) indicates a statistically significant difference ( P ≤.05) between groups. The two groups represent biofilms with either T. forsythia ATCC 43037 wild‐type or T. forsythia UB 4 wild‐type. (B, C) Fluorescence in situ hybridization stainings of fixed biofilms showing the localization of ATCC 43037 wild‐type (B) and UB 4 wild‐type (C). Red/yellow: T. forsythia; cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Here a representative area for one disk each is shown with a top view in the left panel and a side view with the biofilm–disk interface directed towards the top view; scale bars 5 μm (B) and 10 μm (C)
    Figure Legend Snippet: Comparison of 10‐species biofilms with two Tannerella forsythia wild‐type strains. (A) Whiskers boxplots (5th to 95th centile) show bacterial numbers determined by quantitative real‐time PCR from three independent experiments. Asterisk (*) indicates a statistically significant difference ( P ≤.05) between groups. The two groups represent biofilms with either T. forsythia ATCC 43037 wild‐type or T. forsythia UB 4 wild‐type. (B, C) Fluorescence in situ hybridization stainings of fixed biofilms showing the localization of ATCC 43037 wild‐type (B) and UB 4 wild‐type (C). Red/yellow: T. forsythia; cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Here a representative area for one disk each is shown with a top view in the left panel and a side view with the biofilm–disk interface directed towards the top view; scale bars 5 μm (B) and 10 μm (C)

    Techniques Used: Real-time Polymerase Chain Reaction, Fluorescence, In Situ Hybridization, Staining

    Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)
    Figure Legend Snippet: Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)

    Techniques Used: Mutagenesis

    8) Product Images from "Tannerella forsythia strains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications"

    Article Title: Tannerella forsythia strains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications

    Journal: Glycobiology

    doi: 10.1093/glycob/cww129

    Knockout of pseC and legC decreases biofilm formation of T. forsythia ATCC 43037 and T. forsythia UB4 cells, respectively. ( A ) Biofilm formation in T. forsythia ATCC 43037 wild-type compared to ATCC 43037 Δ pseC and the complemented strain ATCC 43037 Δ pseC comp . ( B ) Biofilm formation in T. forsythia UB4 wild-type compared to UB4 Δ legC and the complemented strain T. forsythia UB4 Δ legC comp . Data represent mean values ± SD of at least four independent experiments with four replicates each and were analyzed by the unpaired Student’s t-test. Asterisks indicate significant differences (** P
    Figure Legend Snippet: Knockout of pseC and legC decreases biofilm formation of T. forsythia ATCC 43037 and T. forsythia UB4 cells, respectively. ( A ) Biofilm formation in T. forsythia ATCC 43037 wild-type compared to ATCC 43037 Δ pseC and the complemented strain ATCC 43037 Δ pseC comp . ( B ) Biofilm formation in T. forsythia UB4 wild-type compared to UB4 Δ legC and the complemented strain T. forsythia UB4 Δ legC comp . Data represent mean values ± SD of at least four independent experiments with four replicates each and were analyzed by the unpaired Student’s t-test. Asterisks indicate significant differences (** P

    Techniques Used: Knock-Out

    Experimentally confirmed enzymatic steps (in green) in T. forsythia strains ATCC 43037 (left) and FDC 92A2/UB4 (right) corresponding to the CMP-Pse and CMP-Leg biosynthetic pathways as elucidated in H. pylori and C. jejuni , respectively ( Schoenhofen et al. 2006b , Schoenhofen et al. 2009 ). In T. forsythia , pathways will deviate at some point to produce the unique NulO derivatives found in our strains, such as Pse5Am7Gra. These deviations would be anticipated to occur either within the NulO biosynthetic pathway or post CMP-NulO biosynthesis. Red, blue and orange highlight enzymatic steps that introduce the stereochemical differences between the two pathways and also indicate the positions altered for both hexose intermediates and final NulO. The assignment of roman numerals to each compound is consistent with label designations found throughout the text. Subscripts “P” and “L” indicate intermediates from the CMP-Pse and CMP-Leg biosynthesis pathway, respectively. For simplicity, all sugars except for the NulOs are shown in 4 C 1 form. This figure is available in black and white in print and in color at Glycobiology online.
    Figure Legend Snippet: Experimentally confirmed enzymatic steps (in green) in T. forsythia strains ATCC 43037 (left) and FDC 92A2/UB4 (right) corresponding to the CMP-Pse and CMP-Leg biosynthetic pathways as elucidated in H. pylori and C. jejuni , respectively ( Schoenhofen et al. 2006b , Schoenhofen et al. 2009 ). In T. forsythia , pathways will deviate at some point to produce the unique NulO derivatives found in our strains, such as Pse5Am7Gra. These deviations would be anticipated to occur either within the NulO biosynthetic pathway or post CMP-NulO biosynthesis. Red, blue and orange highlight enzymatic steps that introduce the stereochemical differences between the two pathways and also indicate the positions altered for both hexose intermediates and final NulO. The assignment of roman numerals to each compound is consistent with label designations found throughout the text. Subscripts “P” and “L” indicate intermediates from the CMP-Pse and CMP-Leg biosynthesis pathway, respectively. For simplicity, all sugars except for the NulOs are shown in 4 C 1 form. This figure is available in black and white in print and in color at Glycobiology online.

    Techniques Used: Introduce

    Phylogenetic clusters of selected microbial NulO synthase homologs. Based on the prediction of NulO sugar type by Lewis et al. (2009) , a distance-based neighbor joining tree calculated from the amino acid sequences of NulO synthase enzymes places Tanf_01240 of T. forsythia type strain ATCC 43037 in a phylogenetic clade of pseudaminic acid synthases, while BFO_1066 of strain FDC 92A2/UB4 is clustered with Leg synthases (bootstrap values shown at relevant nodes). Green denotes Leg synthases, red Neu synthases and blue Pse synthases. This figure is available in black and white in print and in color at Glycobiology online.
    Figure Legend Snippet: Phylogenetic clusters of selected microbial NulO synthase homologs. Based on the prediction of NulO sugar type by Lewis et al. (2009) , a distance-based neighbor joining tree calculated from the amino acid sequences of NulO synthase enzymes places Tanf_01240 of T. forsythia type strain ATCC 43037 in a phylogenetic clade of pseudaminic acid synthases, while BFO_1066 of strain FDC 92A2/UB4 is clustered with Leg synthases (bootstrap values shown at relevant nodes). Green denotes Leg synthases, red Neu synthases and blue Pse synthases. This figure is available in black and white in print and in color at Glycobiology online.

    Techniques Used:

    SDS-PAGE gels (12%) of purified recombinant enzymes of the ( A ) CMP-Pse biosynthesis pathway from T. forsythia ATCC 43037 and ( B ) CMP-Leg biosynthesis pathway from T. forsythia FDC 92A2/UB4.
    Figure Legend Snippet: SDS-PAGE gels (12%) of purified recombinant enzymes of the ( A ) CMP-Pse biosynthesis pathway from T. forsythia ATCC 43037 and ( B ) CMP-Leg biosynthesis pathway from T. forsythia FDC 92A2/UB4.

    Techniques Used: SDS Page, Purification, Recombinant

    SDS-PAGE analysis of T. forsythia parent, NulO-deficient, and complemented strains. ( A ) CBB stained SDS-PAGE (7.5% gel) of whole cell extracts from T. forsythia ATCC 43037, ATCC 43037 Δ pseC and the complemented strain ATCC 43037 Δ pseC comp , and ( B ) T. forsythia UB4 wild-type, UB4 Δ legC and the complemented strain UB4 Δ legC comp . For both T. forsythia ATCC 43037 Δ pseC and UB4 Δ legC , a downshift of the S-layer protein bands (TfsA and TfsB) could be observed, which was reverted in the complemented strains.
    Figure Legend Snippet: SDS-PAGE analysis of T. forsythia parent, NulO-deficient, and complemented strains. ( A ) CBB stained SDS-PAGE (7.5% gel) of whole cell extracts from T. forsythia ATCC 43037, ATCC 43037 Δ pseC and the complemented strain ATCC 43037 Δ pseC comp , and ( B ) T. forsythia UB4 wild-type, UB4 Δ legC and the complemented strain UB4 Δ legC comp . For both T. forsythia ATCC 43037 Δ pseC and UB4 Δ legC , a downshift of the S-layer protein bands (TfsA and TfsB) could be observed, which was reverted in the complemented strains.

    Techniques Used: SDS Page, Staining

    9) Product Images from "Tannerella forsythia strains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications"

    Article Title: Tannerella forsythia strains display different cell-surface nonulosonic acids: biosynthetic pathway characterization and first insight into biological implications

    Journal: Glycobiology

    doi: 10.1093/glycob/cww129

    CE analysis of the reaction products obtained by the incubation of UDP-GlcNAc (I P ) upon sequential addition of PseB-His 6 , forming UDP-2-acetamido-2,6-dideoxy-β- l - arabino -hexos-4-ulose (II P ); PseC-MBP, forming UDP-4-amino-4,6-dideoxy-β-L-AltNAc (III P ); PseH-His 6 , forming UDP-2,4-diacetamido-2,4,6-trideoxy-β- l - altro -pyranose (IV P ) and PseG-MBP, releasing UDP. Lastly, the combined action of PseI-His 6 and H. pylori PseF convert 2,4-diacetamido-2,4,6-trideoxy-β- l - altro -pyranose (V P ) via Pse5,7Ac 2 (VI P ) to CMP-Pse5,7Ac 2 (VII P ). Subscript “P” indicates intermediates from the CMP-Pse biosynthesis pathway.
    Figure Legend Snippet: CE analysis of the reaction products obtained by the incubation of UDP-GlcNAc (I P ) upon sequential addition of PseB-His 6 , forming UDP-2-acetamido-2,6-dideoxy-β- l - arabino -hexos-4-ulose (II P ); PseC-MBP, forming UDP-4-amino-4,6-dideoxy-β-L-AltNAc (III P ); PseH-His 6 , forming UDP-2,4-diacetamido-2,4,6-trideoxy-β- l - altro -pyranose (IV P ) and PseG-MBP, releasing UDP. Lastly, the combined action of PseI-His 6 and H. pylori PseF convert 2,4-diacetamido-2,4,6-trideoxy-β- l - altro -pyranose (V P ) via Pse5,7Ac 2 (VI P ) to CMP-Pse5,7Ac 2 (VII P ). Subscript “P” indicates intermediates from the CMP-Pse biosynthesis pathway.

    Techniques Used: Incubation

    10) Product Images from "Regulation and Molecular Basis of Environmental Muropeptide Uptake and Utilization in Fastidious Oral Anaerobe Tannerella forsythia"

    Article Title: Regulation and Molecular Basis of Environmental Muropeptide Uptake and Utilization in Fastidious Oral Anaerobe Tannerella forsythia

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2017.00648

    Growth of Tannerella forsythia on muropeptides. Growth of T. forsythia wild-type and ΔgppX in M9 liquid medium supplemented with 0.2% muropeptides or 0.2% MurNAc was measured at OD 600 . Results of one out of three independent cultivations with similar outcome are given.
    Figure Legend Snippet: Growth of Tannerella forsythia on muropeptides. Growth of T. forsythia wild-type and ΔgppX in M9 liquid medium supplemented with 0.2% muropeptides or 0.2% MurNAc was measured at OD 600 . Results of one out of three independent cultivations with similar outcome are given.

    Techniques Used:

    11) Product Images from "A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia"

    Article Title: A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia

    Journal: Glycobiology

    doi: 10.1093/glycob/cwx019

    ESI-IT-MS analysis of cellular nucleotide sugar pools from T. forsythia strains. ( A ) CMP-activated Pse5Am7Gra ( m / z 683.3) was detected in the T. forsythia ATCC 43037 wild-type and in the Δ Tanf_01245 mutant, whereas this mass was absent in a Pse biosynthesis deficient strain (Δ pseC ) which served as a negative control. ( B ) In T. forsythia UB4 wild-type and in the Δ TFUB4_00887 mutant, a m / z 654.3 peak was identified, which was attributed to a CMP-activated Leg derivative (CMP-Leg*). This mass is consistent with having Ac and Gc modifications on Leg, based on calculation. Notably, this peak was absent in the Legbiosynthesis deficient strain (Δ legC ) which served as a negative control. Relative peak intensities are given on the y axis.
    Figure Legend Snippet: ESI-IT-MS analysis of cellular nucleotide sugar pools from T. forsythia strains. ( A ) CMP-activated Pse5Am7Gra ( m / z 683.3) was detected in the T. forsythia ATCC 43037 wild-type and in the Δ Tanf_01245 mutant, whereas this mass was absent in a Pse biosynthesis deficient strain (Δ pseC ) which served as a negative control. ( B ) In T. forsythia UB4 wild-type and in the Δ TFUB4_00887 mutant, a m / z 654.3 peak was identified, which was attributed to a CMP-activated Leg derivative (CMP-Leg*). This mass is consistent with having Ac and Gc modifications on Leg, based on calculation. Notably, this peak was absent in the Legbiosynthesis deficient strain (Δ legC ) which served as a negative control. Relative peak intensities are given on the y axis.

    Techniques Used: Mass Spectrometry, Mutagenesis, Negative Control

    12) Product Images from "A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia"

    Article Title: A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia

    Journal: Glycobiology

    doi: 10.1093/glycob/cwx019

    ESI-IT-MS analysis of cellular nucleotide sugar pools from T. forsythia strains. ( A ) CMP-activated Pse5Am7Gra ( m / z 683.3) was detected in the T. forsythia ATCC 43037 wild-type and in the Δ Tanf_01245 mutant, whereas this mass was absent in a Pse biosynthesis deficient strain (Δ pseC ) which served as a negative control. ( B ) In T. forsythia UB4 wild-type and in the Δ TFUB4_00887 mutant, a m / z 654.3 peak was identified, which was attributed to a CMP-activated Leg derivative (CMP-Leg*). This mass is consistent with having Ac and Gc modifications on Leg, based on calculation. Notably, this peak was absent in the Legbiosynthesis deficient strain (Δ legC ) which served as a negative control. Relative peak intensities are given on the y axis.
    Figure Legend Snippet: ESI-IT-MS analysis of cellular nucleotide sugar pools from T. forsythia strains. ( A ) CMP-activated Pse5Am7Gra ( m / z 683.3) was detected in the T. forsythia ATCC 43037 wild-type and in the Δ Tanf_01245 mutant, whereas this mass was absent in a Pse biosynthesis deficient strain (Δ pseC ) which served as a negative control. ( B ) In T. forsythia UB4 wild-type and in the Δ TFUB4_00887 mutant, a m / z 654.3 peak was identified, which was attributed to a CMP-activated Leg derivative (CMP-Leg*). This mass is consistent with having Ac and Gc modifications on Leg, based on calculation. Notably, this peak was absent in the Legbiosynthesis deficient strain (Δ legC ) which served as a negative control. Relative peak intensities are given on the y axis.

    Techniques Used: Mass Spectrometry, Mutagenesis, Negative Control

    13) Product Images from "A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia"

    Article Title: A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia

    Journal: Glycobiology

    doi: 10.1093/glycob/cwx019

    ESI-IT-MS analysis of cellular nucleotide sugar pools from T. forsythia strains. ( A ) CMP-activated Pse5Am7Gra ( m / z 683.3) was detected in the T. forsythia ATCC 43037 wild-type and in the Δ Tanf_01245 mutant, whereas this mass was absent in a Pse biosynthesis deficient strain (Δ pseC ) which served as a negative control. ( B ) In T. forsythia UB4 wild-type and in the Δ TFUB4_00887 mutant, a m / z 654.3 peak was identified, which was attributed to a CMP-activated Leg derivative (CMP-Leg*). This mass is consistent with having Ac and Gc modifications on Leg, based on calculation. Notably, this peak was absent in the Legbiosynthesis deficient strain (Δ legC ) which served as a negative control. Relative peak intensities are given on the y axis.
    Figure Legend Snippet: ESI-IT-MS analysis of cellular nucleotide sugar pools from T. forsythia strains. ( A ) CMP-activated Pse5Am7Gra ( m / z 683.3) was detected in the T. forsythia ATCC 43037 wild-type and in the Δ Tanf_01245 mutant, whereas this mass was absent in a Pse biosynthesis deficient strain (Δ pseC ) which served as a negative control. ( B ) In T. forsythia UB4 wild-type and in the Δ TFUB4_00887 mutant, a m / z 654.3 peak was identified, which was attributed to a CMP-activated Leg derivative (CMP-Leg*). This mass is consistent with having Ac and Gc modifications on Leg, based on calculation. Notably, this peak was absent in the Legbiosynthesis deficient strain (Δ legC ) which served as a negative control. Relative peak intensities are given on the y axis.

    Techniques Used: Mass Spectrometry, Mutagenesis, Negative Control

    14) Product Images from "A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia"

    Article Title: A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia

    Journal: Glycobiology

    doi: 10.1093/glycob/cwx019

    ESI-IT-MS analysis of cellular nucleotide sugar pools from T. forsythia strains. ( A ) CMP-activated Pse5Am7Gra ( m / z 683.3) was detected in the T. forsythia ATCC 43037 wild-type and in the Δ Tanf_01245 mutant, whereas this mass was absent in a Pse biosynthesis deficient strain (Δ pseC ) which served as a negative control. ( B ) In T. forsythia UB4 wild-type and in the Δ TFUB4_00887 mutant, a m / z 654.3 peak was identified, which was attributed to a CMP-activated Leg derivative (CMP-Leg*). This mass is consistent with having Ac and Gc modifications on Leg, based on calculation. Notably, this peak was absent in the Legbiosynthesis deficient strain (Δ legC ) which served as a negative control. Relative peak intensities are given on the y axis.
    Figure Legend Snippet: ESI-IT-MS analysis of cellular nucleotide sugar pools from T. forsythia strains. ( A ) CMP-activated Pse5Am7Gra ( m / z 683.3) was detected in the T. forsythia ATCC 43037 wild-type and in the Δ Tanf_01245 mutant, whereas this mass was absent in a Pse biosynthesis deficient strain (Δ pseC ) which served as a negative control. ( B ) In T. forsythia UB4 wild-type and in the Δ TFUB4_00887 mutant, a m / z 654.3 peak was identified, which was attributed to a CMP-activated Leg derivative (CMP-Leg*). This mass is consistent with having Ac and Gc modifications on Leg, based on calculation. Notably, this peak was absent in the Legbiosynthesis deficient strain (Δ legC ) which served as a negative control. Relative peak intensities are given on the y axis.

    Techniques Used: Mass Spectrometry, Mutagenesis, Negative Control

    15) Product Images from "A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia"

    Article Title: A pseudaminic acid or a legionaminic acid derivative transferase is strain-specifically implicated in the general protein O-glycosylation system of the periodontal pathogen Tannerella forsythia

    Journal: Glycobiology

    doi: 10.1093/glycob/cwx019

    ESI-IT-MS analysis of cellular nucleotide sugar pools from T. forsythia strains. ( A ) CMP-activated Pse5Am7Gra ( m / z 683.3) was detected in the T. forsythia ATCC 43037 wild-type and in the Δ Tanf_01245 mutant, whereas this mass was absent in a Pse biosynthesis deficient strain (Δ pseC ) which served as a negative control. ( B ) In T. forsythia UB4 wild-type and in the Δ TFUB4_00887 mutant, a m / z 654.3 peak was identified, which was attributed to a CMP-activated Leg derivative (CMP-Leg*). This mass is consistent with having Ac and Gc modifications on Leg, based on calculation. Notably, this peak was absent in the Legbiosynthesis deficient strain (Δ legC ) which served as a negative control. Relative peak intensities are given on the y axis.
    Figure Legend Snippet: ESI-IT-MS analysis of cellular nucleotide sugar pools from T. forsythia strains. ( A ) CMP-activated Pse5Am7Gra ( m / z 683.3) was detected in the T. forsythia ATCC 43037 wild-type and in the Δ Tanf_01245 mutant, whereas this mass was absent in a Pse biosynthesis deficient strain (Δ pseC ) which served as a negative control. ( B ) In T. forsythia UB4 wild-type and in the Δ TFUB4_00887 mutant, a m / z 654.3 peak was identified, which was attributed to a CMP-activated Leg derivative (CMP-Leg*). This mass is consistent with having Ac and Gc modifications on Leg, based on calculation. Notably, this peak was absent in the Legbiosynthesis deficient strain (Δ legC ) which served as a negative control. Relative peak intensities are given on the y axis.

    Techniques Used: Mass Spectrometry, Mutagenesis, Negative Control

    SDS-PAGE and Western immunoblot analyses of T. forsythia ATCC 43037 and T. forsythia UB4 wild-type and mutants. ( A ) CBB staining of crude cell extracts from T. forsythia ATCC 43037 wild-type, Δ Tanf_01245 mutant, reconstituted mutant Δ Tanf_01245 + and cross-complemented mutant Δ Tanf_01245 + TFUB4_00887 after separation on a 7.5% SDS-PA gel. The S-layer glycoproteins (labeled TfsA and TfsB) are indicated and the down-shift resulting from the loss of the Pse5Am7Gra residue can be observed in the deletion mutant and in the cross-complemented mutant, while in the reconstituted strain the bands are up-shifted again to wild-type level. The same migration profiles could be observed for T. forsythia UB4 wild-type, Δ TFUB4_00887 mutant, reconstituted mutant Δ TFUB4_00887 + and cross-complemented mutant Δ TFUB4_00887 + Tanf_01245 . PageRuler Plus Prestained Protein Ladder (Thermo Fisher Scientific) was used as a protein molecular weight marker. The S-layer glycoprotein bands were further processed for MS analyses. Western immunoblots probed with anti-TfsA antiserum ( B ) and anti-TfsB antiserum ( C ) confirmed the identity of the S-layer glycoproteins in all analyzed T. forsythia species. PageRuler Prestained Protein Ladder (Thermo Fisher Scientific) was used as a molecular weight marker.
    Figure Legend Snippet: SDS-PAGE and Western immunoblot analyses of T. forsythia ATCC 43037 and T. forsythia UB4 wild-type and mutants. ( A ) CBB staining of crude cell extracts from T. forsythia ATCC 43037 wild-type, Δ Tanf_01245 mutant, reconstituted mutant Δ Tanf_01245 + and cross-complemented mutant Δ Tanf_01245 + TFUB4_00887 after separation on a 7.5% SDS-PA gel. The S-layer glycoproteins (labeled TfsA and TfsB) are indicated and the down-shift resulting from the loss of the Pse5Am7Gra residue can be observed in the deletion mutant and in the cross-complemented mutant, while in the reconstituted strain the bands are up-shifted again to wild-type level. The same migration profiles could be observed for T. forsythia UB4 wild-type, Δ TFUB4_00887 mutant, reconstituted mutant Δ TFUB4_00887 + and cross-complemented mutant Δ TFUB4_00887 + Tanf_01245 . PageRuler Plus Prestained Protein Ladder (Thermo Fisher Scientific) was used as a protein molecular weight marker. The S-layer glycoprotein bands were further processed for MS analyses. Western immunoblots probed with anti-TfsA antiserum ( B ) and anti-TfsB antiserum ( C ) confirmed the identity of the S-layer glycoproteins in all analyzed T. forsythia species. PageRuler Prestained Protein Ladder (Thermo Fisher Scientific) was used as a molecular weight marker.

    Techniques Used: SDS Page, Western Blot, Staining, Mutagenesis, Labeling, Migration, Molecular Weight, Marker, Mass Spectrometry

    16) Product Images from "A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications"

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.02008

    Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.
    Figure Legend Snippet: Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.

    Techniques Used: Sequencing

    Effects of protein O -glycosylation on T. forsythia immunogenicity. (A) Secretion of inflammatory cytokines by human DCs upon stimulation with T. forsythia wild-type (WT) and glycosyltransferase-deficient mutants ( T. forsythia Δ gtfE , Δ gtfI , and Δ gtfS ) as measured in culture supernatants by ProcartaPlex Multiplex Immunoassay. (B,C) T cell-priming upon APC stimulation with T. forsythia wild-type and glycosyltransferase-deficient mutants was assessed by culturing PBMCs. (B) T cell activation was measured via expression of CD25 by flow cytometry. Cells were pre-gated for live CD3 + cells, T cell proliferation was assessed after 8 days by CFSE dilution,. (C) CD4 + T cell differentiation was assessed by expression of signature transcription factors for Th17 (RORγT) and Treg (FoxP3) cells as measured by flow cytometry. All data are presented as mean ± SEM of triplicate determinations. One representative out of three (A) and two (B,C) , respectively, independent experiments is shown. Statistically significant differences in (A) are indicated as ∗ p
    Figure Legend Snippet: Effects of protein O -glycosylation on T. forsythia immunogenicity. (A) Secretion of inflammatory cytokines by human DCs upon stimulation with T. forsythia wild-type (WT) and glycosyltransferase-deficient mutants ( T. forsythia Δ gtfE , Δ gtfI , and Δ gtfS ) as measured in culture supernatants by ProcartaPlex Multiplex Immunoassay. (B,C) T cell-priming upon APC stimulation with T. forsythia wild-type and glycosyltransferase-deficient mutants was assessed by culturing PBMCs. (B) T cell activation was measured via expression of CD25 by flow cytometry. Cells were pre-gated for live CD3 + cells, T cell proliferation was assessed after 8 days by CFSE dilution,. (C) CD4 + T cell differentiation was assessed by expression of signature transcription factors for Th17 (RORγT) and Treg (FoxP3) cells as measured by flow cytometry. All data are presented as mean ± SEM of triplicate determinations. One representative out of three (A) and two (B,C) , respectively, independent experiments is shown. Statistically significant differences in (A) are indicated as ∗ p

    Techniques Used: Multiplex Assay, Activation Assay, Expressing, Flow Cytometry, Cytometry, Cell Differentiation

    (A) Scheme of the T. forsythia ATCC 43037 S-layer O -glycan structure. Monosaccharide symbols are shown according to the Symbol Nomenclature for Glycans (SNFG) ( Varki et al., 2015 ). Please note that the position of the branching Fuc remained unclear ( Posch et al., 2011 ) until it was determined in the course of this study to be on the reducing-end Gal. (B) Scheme of the 27-kb protein O -glycosylation gene cluster of T. forsythia ATCC 43037. Wzx (black), flippase; pseBCFHGI (green), CMP-Pse biosynthesis genes; gtfSMILE (blue), Gtf genes; mtfJOY (yellow), Mtf genes; asnB (putative asparagine synthetase B), wecC (UDP- N -acetyl- D -mannosamine dehydrogenase) and wecB (UDP- N- acetylglucosamine 2-epimerase) (purple); hypothetical proteins, HP (gray). Genes are not drawn to scale.
    Figure Legend Snippet: (A) Scheme of the T. forsythia ATCC 43037 S-layer O -glycan structure. Monosaccharide symbols are shown according to the Symbol Nomenclature for Glycans (SNFG) ( Varki et al., 2015 ). Please note that the position of the branching Fuc remained unclear ( Posch et al., 2011 ) until it was determined in the course of this study to be on the reducing-end Gal. (B) Scheme of the 27-kb protein O -glycosylation gene cluster of T. forsythia ATCC 43037. Wzx (black), flippase; pseBCFHGI (green), CMP-Pse biosynthesis genes; gtfSMILE (blue), Gtf genes; mtfJOY (yellow), Mtf genes; asnB (putative asparagine synthetase B), wecC (UDP- N -acetyl- D -mannosamine dehydrogenase) and wecB (UDP- N- acetylglucosamine 2-epimerase) (purple); hypothetical proteins, HP (gray). Genes are not drawn to scale.

    Techniques Used:

    17) Product Images from "A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications"

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.02008

    Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.
    Figure Legend Snippet: Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.

    Techniques Used: Sequencing

    Effects of protein O -glycosylation on T. forsythia immunogenicity. (A) Secretion of inflammatory cytokines by human DCs upon stimulation with T. forsythia wild-type (WT) and glycosyltransferase-deficient mutants ( T. forsythia Δ gtfE , Δ gtfI , and Δ gtfS ) as measured in culture supernatants by ProcartaPlex Multiplex Immunoassay. (B,C) T cell-priming upon APC stimulation with T. forsythia wild-type and glycosyltransferase-deficient mutants was assessed by culturing PBMCs. (B) T cell activation was measured via expression of CD25 by flow cytometry. Cells were pre-gated for live CD3 + cells, T cell proliferation was assessed after 8 days by CFSE dilution,. (C) CD4 + T cell differentiation was assessed by expression of signature transcription factors for Th17 (RORγT) and Treg (FoxP3) cells as measured by flow cytometry. All data are presented as mean ± SEM of triplicate determinations. One representative out of three (A) and two (B,C) , respectively, independent experiments is shown. Statistically significant differences in (A) are indicated as ∗ p
    Figure Legend Snippet: Effects of protein O -glycosylation on T. forsythia immunogenicity. (A) Secretion of inflammatory cytokines by human DCs upon stimulation with T. forsythia wild-type (WT) and glycosyltransferase-deficient mutants ( T. forsythia Δ gtfE , Δ gtfI , and Δ gtfS ) as measured in culture supernatants by ProcartaPlex Multiplex Immunoassay. (B,C) T cell-priming upon APC stimulation with T. forsythia wild-type and glycosyltransferase-deficient mutants was assessed by culturing PBMCs. (B) T cell activation was measured via expression of CD25 by flow cytometry. Cells were pre-gated for live CD3 + cells, T cell proliferation was assessed after 8 days by CFSE dilution,. (C) CD4 + T cell differentiation was assessed by expression of signature transcription factors for Th17 (RORγT) and Treg (FoxP3) cells as measured by flow cytometry. All data are presented as mean ± SEM of triplicate determinations. One representative out of three (A) and two (B,C) , respectively, independent experiments is shown. Statistically significant differences in (A) are indicated as ∗ p

    Techniques Used: Multiplex Assay, Activation Assay, Expressing, Flow Cytometry, Cytometry, Cell Differentiation

    (A) Scheme of the T. forsythia ATCC 43037 S-layer O -glycan structure. Monosaccharide symbols are shown according to the Symbol Nomenclature for Glycans (SNFG) ( Varki et al., 2015 ). Please note that the position of the branching Fuc remained unclear ( Posch et al., 2011 ) until it was determined in the course of this study to be on the reducing-end Gal. (B) Scheme of the 27-kb protein O -glycosylation gene cluster of T. forsythia ATCC 43037. Wzx (black), flippase; pseBCFHGI (green), CMP-Pse biosynthesis genes; gtfSMILE (blue), Gtf genes; mtfJOY (yellow), Mtf genes; asnB (putative asparagine synthetase B), wecC (UDP- N -acetyl- D -mannosamine dehydrogenase) and wecB (UDP- N- acetylglucosamine 2-epimerase) (purple); hypothetical proteins, HP (gray). Genes are not drawn to scale.
    Figure Legend Snippet: (A) Scheme of the T. forsythia ATCC 43037 S-layer O -glycan structure. Monosaccharide symbols are shown according to the Symbol Nomenclature for Glycans (SNFG) ( Varki et al., 2015 ). Please note that the position of the branching Fuc remained unclear ( Posch et al., 2011 ) until it was determined in the course of this study to be on the reducing-end Gal. (B) Scheme of the 27-kb protein O -glycosylation gene cluster of T. forsythia ATCC 43037. Wzx (black), flippase; pseBCFHGI (green), CMP-Pse biosynthesis genes; gtfSMILE (blue), Gtf genes; mtfJOY (yellow), Mtf genes; asnB (putative asparagine synthetase B), wecC (UDP- N -acetyl- D -mannosamine dehydrogenase) and wecB (UDP- N- acetylglucosamine 2-epimerase) (purple); hypothetical proteins, HP (gray). Genes are not drawn to scale.

    Techniques Used:

    18) Product Images from "A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications"

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.02008

    Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.
    Figure Legend Snippet: Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.

    Techniques Used: Sequencing

    Effects of protein O -glycosylation on T. forsythia immunogenicity. (A) Secretion of inflammatory cytokines by human DCs upon stimulation with T. forsythia wild-type (WT) and glycosyltransferase-deficient mutants ( T. forsythia Δ gtfE , Δ gtfI , and Δ gtfS ) as measured in culture supernatants by ProcartaPlex Multiplex Immunoassay. (B,C) T cell-priming upon APC stimulation with T. forsythia wild-type and glycosyltransferase-deficient mutants was assessed by culturing PBMCs. (B) T cell activation was measured via expression of CD25 by flow cytometry. Cells were pre-gated for live CD3 + cells, T cell proliferation was assessed after 8 days by CFSE dilution,. (C) CD4 + T cell differentiation was assessed by expression of signature transcription factors for Th17 (RORγT) and Treg (FoxP3) cells as measured by flow cytometry. All data are presented as mean ± SEM of triplicate determinations. One representative out of three (A) and two (B,C) , respectively, independent experiments is shown. Statistically significant differences in (A) are indicated as ∗ p
    Figure Legend Snippet: Effects of protein O -glycosylation on T. forsythia immunogenicity. (A) Secretion of inflammatory cytokines by human DCs upon stimulation with T. forsythia wild-type (WT) and glycosyltransferase-deficient mutants ( T. forsythia Δ gtfE , Δ gtfI , and Δ gtfS ) as measured in culture supernatants by ProcartaPlex Multiplex Immunoassay. (B,C) T cell-priming upon APC stimulation with T. forsythia wild-type and glycosyltransferase-deficient mutants was assessed by culturing PBMCs. (B) T cell activation was measured via expression of CD25 by flow cytometry. Cells were pre-gated for live CD3 + cells, T cell proliferation was assessed after 8 days by CFSE dilution,. (C) CD4 + T cell differentiation was assessed by expression of signature transcription factors for Th17 (RORγT) and Treg (FoxP3) cells as measured by flow cytometry. All data are presented as mean ± SEM of triplicate determinations. One representative out of three (A) and two (B,C) , respectively, independent experiments is shown. Statistically significant differences in (A) are indicated as ∗ p

    Techniques Used: Multiplex Assay, Activation Assay, Expressing, Flow Cytometry, Cytometry, Cell Differentiation

    (A) Scheme of the T. forsythia ATCC 43037 S-layer O -glycan structure. Monosaccharide symbols are shown according to the Symbol Nomenclature for Glycans (SNFG) ( Varki et al., 2015 ). Please note that the position of the branching Fuc remained unclear ( Posch et al., 2011 ) until it was determined in the course of this study to be on the reducing-end Gal. (B) Scheme of the 27-kb protein O -glycosylation gene cluster of T. forsythia ATCC 43037. Wzx (black), flippase; pseBCFHGI (green), CMP-Pse biosynthesis genes; gtfSMILE (blue), Gtf genes; mtfJOY (yellow), Mtf genes; asnB (putative asparagine synthetase B), wecC (UDP- N -acetyl- D -mannosamine dehydrogenase) and wecB (UDP- N- acetylglucosamine 2-epimerase) (purple); hypothetical proteins, HP (gray). Genes are not drawn to scale.
    Figure Legend Snippet: (A) Scheme of the T. forsythia ATCC 43037 S-layer O -glycan structure. Monosaccharide symbols are shown according to the Symbol Nomenclature for Glycans (SNFG) ( Varki et al., 2015 ). Please note that the position of the branching Fuc remained unclear ( Posch et al., 2011 ) until it was determined in the course of this study to be on the reducing-end Gal. (B) Scheme of the 27-kb protein O -glycosylation gene cluster of T. forsythia ATCC 43037. Wzx (black), flippase; pseBCFHGI (green), CMP-Pse biosynthesis genes; gtfSMILE (blue), Gtf genes; mtfJOY (yellow), Mtf genes; asnB (putative asparagine synthetase B), wecC (UDP- N -acetyl- D -mannosamine dehydrogenase) and wecB (UDP- N- acetylglucosamine 2-epimerase) (purple); hypothetical proteins, HP (gray). Genes are not drawn to scale.

    Techniques Used:

    19) Product Images from "Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia" "

    Article Title: Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia"

    Journal:

    doi: 10.1128/IAI.00062-06

    Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. gingivalis OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number
    Figure Legend Snippet: Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. gingivalis OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number

    Techniques Used:

    Coaggregation of T. forsythia in the presence of P. gingivalis and outer membrane vesicles purified from P. gingivalis . Data are representative of three independent experiments. Data points represent means ± standard errors of triplicate readings.
    Figure Legend Snippet: Coaggregation of T. forsythia in the presence of P. gingivalis and outer membrane vesicles purified from P. gingivalis . Data are representative of three independent experiments. Data points represent means ± standard errors of triplicate readings.

    Techniques Used: Purification

    Invasion by T. forsythia (Tf) of KB cells and dependence of invasion on the presence of P. gingivalis vesicles (Pgv). The invasion level was expressed as the percentage of bacteria retrieved following antibiotic killing of external bacteria and KB cell
    Figure Legend Snippet: Invasion by T. forsythia (Tf) of KB cells and dependence of invasion on the presence of P. gingivalis vesicles (Pgv). The invasion level was expressed as the percentage of bacteria retrieved following antibiotic killing of external bacteria and KB cell

    Techniques Used:

    20) Product Images from "Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia" "

    Article Title: Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia"

    Journal:

    doi: 10.1128/IAI.00062-06

    Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. gingivalis OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number
    Figure Legend Snippet: Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. gingivalis OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number

    Techniques Used:

    Coaggregation of T. forsythia in the presence of P. gingivalis and outer membrane vesicles purified from P. gingivalis . Data are representative of three independent experiments. Data points represent means ± standard errors of triplicate readings.
    Figure Legend Snippet: Coaggregation of T. forsythia in the presence of P. gingivalis and outer membrane vesicles purified from P. gingivalis . Data are representative of three independent experiments. Data points represent means ± standard errors of triplicate readings.

    Techniques Used: Purification

    21) Product Images from "N-Acetylmuramic Acid (MurNAc) Auxotrophy of the Oral Pathogen Tannerella forsythia: Characterization of a MurNAc Kinase and Analysis of Its Role in Cell Wall Metabolism"

    Article Title: N-Acetylmuramic Acid (MurNAc) Auxotrophy of the Oral Pathogen Tannerella forsythia: Characterization of a MurNAc Kinase and Analysis of Its Role in Cell Wall Metabolism

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.00019

    The rTf_MurK reaction product is MurNAc-6-phosphate (MurNAc-6P). The product of the rTf_MurK kinase reaction with the substrate MurNAc, MurNAc-6P (red) (A) was degraded by the specific MurNAc-6P etherase of T. forsythia (rTf_MurQ) yielding the reaction product GlcNAc-6P (blue) as followed by LC-MS (B) . Shown are the extracted ion chromatograms (EICs) in negative ion mode: (M-H) - = 372.072 m/z for MurNAc-6P (red) at a retention time of 22 min and (M-H) - = 300.059 m/z for GlcNAc-6P (blue) at a retention time of 10 min on a Gemini C-18 RP-column.
    Figure Legend Snippet: The rTf_MurK reaction product is MurNAc-6-phosphate (MurNAc-6P). The product of the rTf_MurK kinase reaction with the substrate MurNAc, MurNAc-6P (red) (A) was degraded by the specific MurNAc-6P etherase of T. forsythia (rTf_MurQ) yielding the reaction product GlcNAc-6P (blue) as followed by LC-MS (B) . Shown are the extracted ion chromatograms (EICs) in negative ion mode: (M-H) - = 372.072 m/z for MurNAc-6P (red) at a retention time of 22 min and (M-H) - = 300.059 m/z for GlcNAc-6P (blue) at a retention time of 10 min on a Gemini C-18 RP-column.

    Techniques Used: Liquid Chromatography with Mass Spectroscopy

    22) Product Images from "A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications"

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.02008

    Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.
    Figure Legend Snippet: Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.

    Techniques Used: Sequencing

    Effects of protein O -glycosylation on T. forsythia immunogenicity. (A) Secretion of inflammatory cytokines by human DCs upon stimulation with T. forsythia wild-type (WT) and glycosyltransferase-deficient mutants ( T. forsythia Δ gtfE , Δ gtfI , and Δ gtfS ) as measured in culture supernatants by ProcartaPlex Multiplex Immunoassay. (B,C) T cell-priming upon APC stimulation with T. forsythia wild-type and glycosyltransferase-deficient mutants was assessed by culturing PBMCs. (B) T cell activation was measured via expression of CD25 by flow cytometry. Cells were pre-gated for live CD3 + cells, T cell proliferation was assessed after 8 days by CFSE dilution,. (C) CD4 + T cell differentiation was assessed by expression of signature transcription factors for Th17 (RORγT) and Treg (FoxP3) cells as measured by flow cytometry. All data are presented as mean ± SEM of triplicate determinations. One representative out of three (A) and two (B,C) , respectively, independent experiments is shown. Statistically significant differences in (A) are indicated as ∗ p
    Figure Legend Snippet: Effects of protein O -glycosylation on T. forsythia immunogenicity. (A) Secretion of inflammatory cytokines by human DCs upon stimulation with T. forsythia wild-type (WT) and glycosyltransferase-deficient mutants ( T. forsythia Δ gtfE , Δ gtfI , and Δ gtfS ) as measured in culture supernatants by ProcartaPlex Multiplex Immunoassay. (B,C) T cell-priming upon APC stimulation with T. forsythia wild-type and glycosyltransferase-deficient mutants was assessed by culturing PBMCs. (B) T cell activation was measured via expression of CD25 by flow cytometry. Cells were pre-gated for live CD3 + cells, T cell proliferation was assessed after 8 days by CFSE dilution,. (C) CD4 + T cell differentiation was assessed by expression of signature transcription factors for Th17 (RORγT) and Treg (FoxP3) cells as measured by flow cytometry. All data are presented as mean ± SEM of triplicate determinations. One representative out of three (A) and two (B,C) , respectively, independent experiments is shown. Statistically significant differences in (A) are indicated as ∗ p

    Techniques Used: Multiplex Assay, Activation Assay, Expressing, Flow Cytometry, Cytometry, Cell Differentiation

    23) Product Images from "A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications"

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.02008

    Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.
    Figure Legend Snippet: Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.

    Techniques Used: Sequencing

    Effects of protein O -glycosylation on T. forsythia immunogenicity. (A) Secretion of inflammatory cytokines by human DCs upon stimulation with T. forsythia wild-type (WT) and glycosyltransferase-deficient mutants ( T. forsythia Δ gtfE , Δ gtfI , and Δ gtfS ) as measured in culture supernatants by ProcartaPlex Multiplex Immunoassay. (B,C) T cell-priming upon APC stimulation with T. forsythia wild-type and glycosyltransferase-deficient mutants was assessed by culturing PBMCs. (B) T cell activation was measured via expression of CD25 by flow cytometry. Cells were pre-gated for live CD3 + cells, T cell proliferation was assessed after 8 days by CFSE dilution,. (C) CD4 + T cell differentiation was assessed by expression of signature transcription factors for Th17 (RORγT) and Treg (FoxP3) cells as measured by flow cytometry. All data are presented as mean ± SEM of triplicate determinations. One representative out of three (A) and two (B,C) , respectively, independent experiments is shown. Statistically significant differences in (A) are indicated as ∗ p
    Figure Legend Snippet: Effects of protein O -glycosylation on T. forsythia immunogenicity. (A) Secretion of inflammatory cytokines by human DCs upon stimulation with T. forsythia wild-type (WT) and glycosyltransferase-deficient mutants ( T. forsythia Δ gtfE , Δ gtfI , and Δ gtfS ) as measured in culture supernatants by ProcartaPlex Multiplex Immunoassay. (B,C) T cell-priming upon APC stimulation with T. forsythia wild-type and glycosyltransferase-deficient mutants was assessed by culturing PBMCs. (B) T cell activation was measured via expression of CD25 by flow cytometry. Cells were pre-gated for live CD3 + cells, T cell proliferation was assessed after 8 days by CFSE dilution,. (C) CD4 + T cell differentiation was assessed by expression of signature transcription factors for Th17 (RORγT) and Treg (FoxP3) cells as measured by flow cytometry. All data are presented as mean ± SEM of triplicate determinations. One representative out of three (A) and two (B,C) , respectively, independent experiments is shown. Statistically significant differences in (A) are indicated as ∗ p

    Techniques Used: Multiplex Assay, Activation Assay, Expressing, Flow Cytometry, Cytometry, Cell Differentiation

    24) Product Images from "Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms"

    Article Title: Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

    Journal: Molecular Oral Microbiology

    doi: 10.1111/omi.12182

    Dual fluorescence in situ hybridization staining of Tannerella forsythia and Campylobacter rectus for biofilms harboring ATCC 43037 wild‐type (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: T. forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)
    Figure Legend Snippet: Dual fluorescence in situ hybridization staining of Tannerella forsythia and Campylobacter rectus for biofilms harboring ATCC 43037 wild‐type (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: T. forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)

    Techniques Used: Fluorescence, In Situ Hybridization, Staining

    Comparison of colony‐forming unit ( CFU ) counting and quantitative polymerase chain reaction ( qPCR ) for Tannerella forsythia wild‐type strains and mutants in the subgingival “Zurich biofilm”. Total bacteria for 10‐species biofilms with different T. forsythia strains and mutants enumerated by CFU counts (red boxes) and qPCR (blue boxes) for three independent experiments with three technical replicates, each, are shown (Whiskers boxplots 5th to 95th centile)
    Figure Legend Snippet: Comparison of colony‐forming unit ( CFU ) counting and quantitative polymerase chain reaction ( qPCR ) for Tannerella forsythia wild‐type strains and mutants in the subgingival “Zurich biofilm”. Total bacteria for 10‐species biofilms with different T. forsythia strains and mutants enumerated by CFU counts (red boxes) and qPCR (blue boxes) for three independent experiments with three technical replicates, each, are shown (Whiskers boxplots 5th to 95th centile)

    Techniques Used: Real-time Polymerase Chain Reaction

    Comparison of 10‐species biofilms with two Tannerella forsythia wild‐type strains. (A) Whiskers boxplots (5th to 95th centile) show bacterial numbers determined by quantitative real‐time PCR from three independent experiments. Asterisk (*) indicates a statistically significant difference ( P ≤.05) between groups. The two groups represent biofilms with either T. forsythia ATCC 43037 wild‐type or T. forsythia UB 4 wild‐type. (B, C) Fluorescence in situ hybridization stainings of fixed biofilms showing the localization of ATCC 43037 wild‐type (B) and UB 4 wild‐type (C). Red/yellow: T. forsythia; cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Here a representative area for one disk each is shown with a top view in the left panel and a side view with the biofilm–disk interface directed towards the top view; scale bars 5 μm (B) and 10 μm (C)
    Figure Legend Snippet: Comparison of 10‐species biofilms with two Tannerella forsythia wild‐type strains. (A) Whiskers boxplots (5th to 95th centile) show bacterial numbers determined by quantitative real‐time PCR from three independent experiments. Asterisk (*) indicates a statistically significant difference ( P ≤.05) between groups. The two groups represent biofilms with either T. forsythia ATCC 43037 wild‐type or T. forsythia UB 4 wild‐type. (B, C) Fluorescence in situ hybridization stainings of fixed biofilms showing the localization of ATCC 43037 wild‐type (B) and UB 4 wild‐type (C). Red/yellow: T. forsythia; cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Here a representative area for one disk each is shown with a top view in the left panel and a side view with the biofilm–disk interface directed towards the top view; scale bars 5 μm (B) and 10 μm (C)

    Techniques Used: Real-time Polymerase Chain Reaction, Fluorescence, In Situ Hybridization, Staining

    Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)
    Figure Legend Snippet: Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)

    Techniques Used: Mutagenesis

    25) Product Images from "Comparative genome characterization of the periodontal pathogen Tannerella forsythia"

    Article Title: Comparative genome characterization of the periodontal pathogen Tannerella forsythia

    Journal: BMC Genomics

    doi: 10.1186/s12864-020-6535-y

    Multiple whole genome alignment of eight T. forsythia strains. Each coloured block represents a genomic region that aligned to a region in at least one other genome, plotted in the same colour, to which it was predicted to be homologous based on sequence similarity. Blocks above the centre line indicate forward orientation; blocks below the line indicate reverse orientation relative to strain 92A2. A histogram within each block shows the average similarity of a region to its counterparts in the other genomes. Red vertical lines indicate contig boundaries. Strain ATCC 43037 displayed two translocations compared to strain 92A2 with lengths of approximately 500 kbp (blue and yellow blocks at the right end of 92A2 and in the centre of ATCC) and 30 kbp (pink block at approx. 1.25 Mbp in 92A2 and at approx. 2.7 Mbp in ATCC), respectively. Previously described large-scale inversions in strain KS16 could be confirmed (reverted blocks in the left half of the alignment)
    Figure Legend Snippet: Multiple whole genome alignment of eight T. forsythia strains. Each coloured block represents a genomic region that aligned to a region in at least one other genome, plotted in the same colour, to which it was predicted to be homologous based on sequence similarity. Blocks above the centre line indicate forward orientation; blocks below the line indicate reverse orientation relative to strain 92A2. A histogram within each block shows the average similarity of a region to its counterparts in the other genomes. Red vertical lines indicate contig boundaries. Strain ATCC 43037 displayed two translocations compared to strain 92A2 with lengths of approximately 500 kbp (blue and yellow blocks at the right end of 92A2 and in the centre of ATCC) and 30 kbp (pink block at approx. 1.25 Mbp in 92A2 and at approx. 2.7 Mbp in ATCC), respectively. Previously described large-scale inversions in strain KS16 could be confirmed (reverted blocks in the left half of the alignment)

    Techniques Used: Blocking Assay, Sequencing

    26) Product Images from "Free Lipid A Isolated from Porphyromonas gingivalis Lipopolysaccharide Is Contaminated with Phosphorylated Dihydroceramide Lipids: Recovery in Diseased Dental Samples"

    Article Title: Free Lipid A Isolated from Porphyromonas gingivalis Lipopolysaccharide Is Contaminated with Phosphorylated Dihydroceramide Lipids: Recovery in Diseased Dental Samples

    Journal: Infection and Immunity

    doi: 10.1128/IAI.06180-11

    ESI-MS analysis of P. gingivalis lipid A.
    Figure Legend Snippet: ESI-MS analysis of P. gingivalis lipid A.

    Techniques Used: Mass Spectrometry

    ESI-MS analysis of P. gingivalis lipid A.
    Figure Legend Snippet: ESI-MS analysis of P. gingivalis lipid A.

    Techniques Used: Mass Spectrometry

    Negative and positive ions of lipid A detected by ESI-MS. (A) Shown is the negative-ion spectrum of P. gingivalis lipid A, as demonstrated by ESI-MS. This analysis revealed a substantially different mass spectrum compared with MALDI-MS shown in
    Figure Legend Snippet: Negative and positive ions of lipid A detected by ESI-MS. (A) Shown is the negative-ion spectrum of P. gingivalis lipid A, as demonstrated by ESI-MS. This analysis revealed a substantially different mass spectrum compared with MALDI-MS shown in

    Techniques Used: Mass Spectrometry

    Recovery of phosphorylated dihydroceramide lipids in the lipid A extract of P. gingivalis . Lipid A of P. gingivalis was prepared as described in Materials and Methods, and the lipid A was dissolved in hexane-isopropanol-water (6:8:0.75, vol/vol/vol) at
    Figure Legend Snippet: Recovery of phosphorylated dihydroceramide lipids in the lipid A extract of P. gingivalis . Lipid A of P. gingivalis was prepared as described in Materials and Methods, and the lipid A was dissolved in hexane-isopropanol-water (6:8:0.75, vol/vol/vol) at

    Techniques Used:

    Structure of the high-mass lipid A of P. gingivalis ( m/z 1,690 negative ion) and its ion fragments. (A) This lipid A negative ion is shown with the phosphate group in the 1 position. Shown is the theoretical exact mass (1,689.26 amu) and theoretical molar
    Figure Legend Snippet: Structure of the high-mass lipid A of P. gingivalis ( m/z 1,690 negative ion) and its ion fragments. (A) This lipid A negative ion is shown with the phosphate group in the 1 position. Shown is the theoretical exact mass (1,689.26 amu) and theoretical molar

    Techniques Used:

    MALDI-TOF analysis of P. gingivalis lipid A.
    Figure Legend Snippet: MALDI-TOF analysis of P. gingivalis lipid A.

    Techniques Used:

    Structural reconciliation of the dominant nonphosphorylated lipid A of P. gingivalis . Nonphosphorylated lipid A species are always detected as sodium adducts. (A) Shown is the structure of the most abundant lipid A positive ion ( m/z 1,392.3). The observed
    Figure Legend Snippet: Structural reconciliation of the dominant nonphosphorylated lipid A of P. gingivalis . Nonphosphorylated lipid A species are always detected as sodium adducts. (A) Shown is the structure of the most abundant lipid A positive ion ( m/z 1,392.3). The observed

    Techniques Used:

    27) Product Images from "Porphyromonas endodontalis in chronic periodontitis: a clinical and microbiological cross-sectional study"

    Article Title: Porphyromonas endodontalis in chronic periodontitis: a clinical and microbiological cross-sectional study

    Journal: Journal of Oral Microbiology

    doi: 10.3402/jom.v4i0.10123

    Quantification of P. endodontalis, P. gingivalis and T. forsythia in subgingival plaque of patients with chronic periodontitis by qPCR. Letters represented the evaluating of each bacterium separately before and after periodontal treatment. So those bacteria that had statistically significant reduction after periodontal treatment were represented with different letters.
    Figure Legend Snippet: Quantification of P. endodontalis, P. gingivalis and T. forsythia in subgingival plaque of patients with chronic periodontitis by qPCR. Letters represented the evaluating of each bacterium separately before and after periodontal treatment. So those bacteria that had statistically significant reduction after periodontal treatment were represented with different letters.

    Techniques Used: Real-time Polymerase Chain Reaction

    28) Product Images from "Porphyromonas endodontalis in chronic periodontitis: a clinical and microbiological cross-sectional study"

    Article Title: Porphyromonas endodontalis in chronic periodontitis: a clinical and microbiological cross-sectional study

    Journal: Journal of Oral Microbiology

    doi: 10.3402/jom.v4i0.10123

    Quantification of P. endodontalis, P. gingivalis and T. forsythia in subgingival plaque of patients with chronic periodontitis by qPCR. Letters represented the evaluating of each bacterium separately before and after periodontal treatment. So those bacteria that had statistically significant reduction after periodontal treatment were represented with different letters.
    Figure Legend Snippet: Quantification of P. endodontalis, P. gingivalis and T. forsythia in subgingival plaque of patients with chronic periodontitis by qPCR. Letters represented the evaluating of each bacterium separately before and after periodontal treatment. So those bacteria that had statistically significant reduction after periodontal treatment were represented with different letters.

    Techniques Used: Real-time Polymerase Chain Reaction

    29) Product Images from "Porphyromonas endodontalis in chronic periodontitis: a clinical and microbiological cross-sectional study"

    Article Title: Porphyromonas endodontalis in chronic periodontitis: a clinical and microbiological cross-sectional study

    Journal: Journal of Oral Microbiology

    doi: 10.3402/jom.v4i0.10123

    Quantification of P. endodontalis, P. gingivalis and T. forsythia in subgingival plaque of patients with chronic periodontitis by qPCR. Letters represented the evaluating of each bacterium separately before and after periodontal treatment. So those bacteria that had statistically significant reduction after periodontal treatment were represented with different letters.
    Figure Legend Snippet: Quantification of P. endodontalis, P. gingivalis and T. forsythia in subgingival plaque of patients with chronic periodontitis by qPCR. Letters represented the evaluating of each bacterium separately before and after periodontal treatment. So those bacteria that had statistically significant reduction after periodontal treatment were represented with different letters.

    Techniques Used: Real-time Polymerase Chain Reaction

    30) Product Images from "Antibacterial Activity of As-Annealed TiO2 Nanotubes Doped with Ag Nanoparticles against Periodontal Pathogens"

    Article Title: Antibacterial Activity of As-Annealed TiO2 Nanotubes Doped with Ag Nanoparticles against Periodontal Pathogens

    Journal: Bioinorganic Chemistry and Applications

    doi: 10.1155/2014/829496

    Descriptive analysis of adhesion of (a) A. actinomycetemcomitans , (b) T. forsythia , and (c) C. rectus on all groups tested (Group TiO 2 : as-annealed TiO 2 nanotubes; Group Ag: as-annealed Ag doped TiO 2 nanotubes; Group Ti: commercially pure Ti sheet; Control Group: 24-well cell culture plate bottoms). Data are presented as the mean ± SD (standard deviation). Results were analyzed using a one-way ANOVA and post hoc analyses were performed using Tukey's studentized range (HSD) test (* P
    Figure Legend Snippet: Descriptive analysis of adhesion of (a) A. actinomycetemcomitans , (b) T. forsythia , and (c) C. rectus on all groups tested (Group TiO 2 : as-annealed TiO 2 nanotubes; Group Ag: as-annealed Ag doped TiO 2 nanotubes; Group Ti: commercially pure Ti sheet; Control Group: 24-well cell culture plate bottoms). Data are presented as the mean ± SD (standard deviation). Results were analyzed using a one-way ANOVA and post hoc analyses were performed using Tukey's studentized range (HSD) test (* P

    Techniques Used: Cell Culture, Standard Deviation

    SEM micrographs after adhesion of A. actinomycetemcomitans , T. forsythia , and C. rectus on the surface of (a) Group TiO 2 : as-annealed TiO 2 nanotubes; (b) Group Ag: as-annealed Ag doped TiO 2 nanotubes; (c) Group Ti: commercially pure Ti sheet.
    Figure Legend Snippet: SEM micrographs after adhesion of A. actinomycetemcomitans , T. forsythia , and C. rectus on the surface of (a) Group TiO 2 : as-annealed TiO 2 nanotubes; (b) Group Ag: as-annealed Ag doped TiO 2 nanotubes; (c) Group Ti: commercially pure Ti sheet.

    Techniques Used:

    31) Product Images from "Genome-Wide Relatedness of Treponema pedis, from Gingiva and Necrotic Skin Lesions of Pigs, with the Human Oral Pathogen Treponema denticola"

    Article Title: Genome-Wide Relatedness of Treponema pedis, from Gingiva and Necrotic Skin Lesions of Pigs, with the Human Oral Pathogen Treponema denticola

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0071281

    Circular representation of the T. pedis T A4 genome and complete genome alignment with T. denticola . (A.) Circular representation of the T. pedis T A4 genome. The CDSs are shown in violet where the outer circle represents predictions on the plus strand and the second circle those on the minus strand. CDSs with a best BLASTP hit in T. denticola ATCC 35405 are colored red and shown in the third circle. The fourth circle represents genes with best BLASTP hits in T. brennaborense (black), F. nucleatum (green), F. alocis (blue) and T. succinifaciens (grey). G+C skew is drawn in the inner circle. (B.) Complete genome alignment between T. pedis T A4 and T. denticola ATCC 35405. Dots represent maximum unique matches (MUMs) between the genomes. MUMs oriented in the same direction are depicted as red dots and reverse complemented MUMs are depicted as blue dots.
    Figure Legend Snippet: Circular representation of the T. pedis T A4 genome and complete genome alignment with T. denticola . (A.) Circular representation of the T. pedis T A4 genome. The CDSs are shown in violet where the outer circle represents predictions on the plus strand and the second circle those on the minus strand. CDSs with a best BLASTP hit in T. denticola ATCC 35405 are colored red and shown in the third circle. The fourth circle represents genes with best BLASTP hits in T. brennaborense (black), F. nucleatum (green), F. alocis (blue) and T. succinifaciens (grey). G+C skew is drawn in the inner circle. (B.) Complete genome alignment between T. pedis T A4 and T. denticola ATCC 35405. Dots represent maximum unique matches (MUMs) between the genomes. MUMs oriented in the same direction are depicted as red dots and reverse complemented MUMs are depicted as blue dots.

    Techniques Used:

    32) Product Images from "Impact of aging on TREM-1 responses in the periodontium: a cross-sectional study in an elderly population"

    Article Title: Impact of aging on TREM-1 responses in the periodontium: a cross-sectional study in an elderly population

    Journal: BMC Infectious Diseases

    doi: 10.1186/s12879-016-1778-6

    Oral microbiota levels in the subgingival plaque from elderly individuals with periodontal health ( n = 17), gingivitis ( n = 19), and chronic periodontitis ( n = 15). The individual values represent bacterial counts per sample in the subgingival plaque ( a ) Total bacterial counts, ( b ) F. nucleatum , ( c ) P. intermedia , ( d ) P. gingivalis , ( e ) T. denticola , and ( f ) T. forsythia . The horizontal lines in the middle of the box represent the median and whiskers are drawn down to the 10 th through 90 th percentiles. The points below and above the whiskers are drawn as individual dots
    Figure Legend Snippet: Oral microbiota levels in the subgingival plaque from elderly individuals with periodontal health ( n = 17), gingivitis ( n = 19), and chronic periodontitis ( n = 15). The individual values represent bacterial counts per sample in the subgingival plaque ( a ) Total bacterial counts, ( b ) F. nucleatum , ( c ) P. intermedia , ( d ) P. gingivalis , ( e ) T. denticola , and ( f ) T. forsythia . The horizontal lines in the middle of the box represent the median and whiskers are drawn down to the 10 th through 90 th percentiles. The points below and above the whiskers are drawn as individual dots

    Techniques Used:

    33) Product Images from " Development of In Vitro Denture Biofilm Models for Halitosis Related Bacteria and their Application in Testing the Efficacy of Antimicrobial Agents"

    Article Title: Development of In Vitro Denture Biofilm Models for Halitosis Related Bacteria and their Application in Testing the Efficacy of Antimicrobial Agents

    Journal: The Open Dentistry Journal

    doi: 10.2174/1874210601509010125

    Biofilm formation on DBR disc surfaces. Representative images after crystal violet staining of biofilms formed on DBR discs by F. nucleatum ATCC 23726 (A) T. forsythia ATCC 43037 (B) V. atypica PK 1910 (C) K. pneumoniae IA 565 (D) For each set, the image on the left was a control DBR disc without biofilms growing on the surface, the image on the right was a DBR disc with biofilms growing on the surface. The experiment was performed in triplicate.
    Figure Legend Snippet: Biofilm formation on DBR disc surfaces. Representative images after crystal violet staining of biofilms formed on DBR discs by F. nucleatum ATCC 23726 (A) T. forsythia ATCC 43037 (B) V. atypica PK 1910 (C) K. pneumoniae IA 565 (D) For each set, the image on the left was a control DBR disc without biofilms growing on the surface, the image on the right was a DBR disc with biofilms growing on the surface. The experiment was performed in triplicate.

    Techniques Used: Staining, IA

    CLSM images of biofilms on DBR disc surfaces. Biofilm formation of F. nucleatum ATCC 23726 (A) T. forsythia ATCC 43037 (B) V. atypica PK 1910 (C) K. pneumoniae IA 565 (D) on DBR discs. For each set, the image on the left was taken through a 20x objective (Scale bar, 50 μm); while the image on the right was taken through a 63x objective (Scale bar, 20 μm). Four random fields of view were examined for each sample and representative images are shown.
    Figure Legend Snippet: CLSM images of biofilms on DBR disc surfaces. Biofilm formation of F. nucleatum ATCC 23726 (A) T. forsythia ATCC 43037 (B) V. atypica PK 1910 (C) K. pneumoniae IA 565 (D) on DBR discs. For each set, the image on the left was taken through a 20x objective (Scale bar, 50 μm); while the image on the right was taken through a 63x objective (Scale bar, 20 μm). Four random fields of view were examined for each sample and representative images are shown.

    Techniques Used: Confocal Laser Scanning Microscopy, IA

    34) Product Images from "In vitro Increased Respiratory Activity of Selected Oral Bacteria May Explain Competitive and Collaborative Interactions in the Oral Microbiome"

    Article Title: In vitro Increased Respiratory Activity of Selected Oral Bacteria May Explain Competitive and Collaborative Interactions in the Oral Microbiome

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2017.00235

    Specific groups of metabolites influenced respiratory activity of each bacterial community. Principal Coordinate Analysis assisted to explain the grouping trends in the cluster analysis, and highlighted similarities in the respiratory activity among the selected oral bacteria at 24 h (A) and 48 h (B) . Aa, Aggregatibacter actinomycetemcomitans (ATCC 43718); Fn, Fusobacterium nucleatum (ATCC 10953); Pg, Porphyromonas gingivalis (ATCC 33277); Pi, Prevotella intermedia (ATCC 25611); Sm, Streptococcus mutans (ATCC 25175); Ssob, Streptococcus sobrinus (ATCC 33478); Tf, Tannerella forsythia (ATCC 43037); An, Actinomyces naeslundii (ATCC 51655); Csput, Capnocytophaga sputigena (ATCC 33612); Sgord, Streptococcus gordonii (ATCC 49818); Avisc, Actinomyces viscosus (ATCC 15987); Smitis, Streptococcus mitis (ATCC 49456); Vparv, Veillonella parvula (DSM 2007); Ssang, Streptococcus sanguinis (LMG 14657), and Ssal, Streptococcus salivarius strain TOVE-R.
    Figure Legend Snippet: Specific groups of metabolites influenced respiratory activity of each bacterial community. Principal Coordinate Analysis assisted to explain the grouping trends in the cluster analysis, and highlighted similarities in the respiratory activity among the selected oral bacteria at 24 h (A) and 48 h (B) . Aa, Aggregatibacter actinomycetemcomitans (ATCC 43718); Fn, Fusobacterium nucleatum (ATCC 10953); Pg, Porphyromonas gingivalis (ATCC 33277); Pi, Prevotella intermedia (ATCC 25611); Sm, Streptococcus mutans (ATCC 25175); Ssob, Streptococcus sobrinus (ATCC 33478); Tf, Tannerella forsythia (ATCC 43037); An, Actinomyces naeslundii (ATCC 51655); Csput, Capnocytophaga sputigena (ATCC 33612); Sgord, Streptococcus gordonii (ATCC 49818); Avisc, Actinomyces viscosus (ATCC 15987); Smitis, Streptococcus mitis (ATCC 49456); Vparv, Veillonella parvula (DSM 2007); Ssang, Streptococcus sanguinis (LMG 14657), and Ssal, Streptococcus salivarius strain TOVE-R.

    Techniques Used: Activity Assay

    35) Product Images from "In vitro Increased Respiratory Activity of Selected Oral Bacteria May Explain Competitive and Collaborative Interactions in the Oral Microbiome"

    Article Title: In vitro Increased Respiratory Activity of Selected Oral Bacteria May Explain Competitive and Collaborative Interactions in the Oral Microbiome

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2017.00235

    Specific groups of metabolites influenced respiratory activity of each bacterial community. Principal Coordinate Analysis assisted to explain the grouping trends in the cluster analysis, and highlighted similarities in the respiratory activity among the selected oral bacteria at 24 h (A) and 48 h (B) . Aa, Aggregatibacter actinomycetemcomitans (ATCC 43718); Fn, Fusobacterium nucleatum (ATCC 10953); Pg, Porphyromonas gingivalis (ATCC 33277); Pi, Prevotella intermedia (ATCC 25611); Sm, Streptococcus mutans (ATCC 25175); Ssob, Streptococcus sobrinus (ATCC 33478); Tf, Tannerella forsythia (ATCC 43037); An, Actinomyces naeslundii (ATCC 51655); Csput, Capnocytophaga sputigena (ATCC 33612); Sgord, Streptococcus gordonii (ATCC 49818); Avisc, Actinomyces viscosus (ATCC 15987); Smitis, Streptococcus mitis (ATCC 49456); Vparv, Veillonella parvula (DSM 2007); Ssang, Streptococcus sanguinis (LMG 14657), and Ssal, Streptococcus salivarius strain TOVE-R.
    Figure Legend Snippet: Specific groups of metabolites influenced respiratory activity of each bacterial community. Principal Coordinate Analysis assisted to explain the grouping trends in the cluster analysis, and highlighted similarities in the respiratory activity among the selected oral bacteria at 24 h (A) and 48 h (B) . Aa, Aggregatibacter actinomycetemcomitans (ATCC 43718); Fn, Fusobacterium nucleatum (ATCC 10953); Pg, Porphyromonas gingivalis (ATCC 33277); Pi, Prevotella intermedia (ATCC 25611); Sm, Streptococcus mutans (ATCC 25175); Ssob, Streptococcus sobrinus (ATCC 33478); Tf, Tannerella forsythia (ATCC 43037); An, Actinomyces naeslundii (ATCC 51655); Csput, Capnocytophaga sputigena (ATCC 33612); Sgord, Streptococcus gordonii (ATCC 49818); Avisc, Actinomyces viscosus (ATCC 15987); Smitis, Streptococcus mitis (ATCC 49456); Vparv, Veillonella parvula (DSM 2007); Ssang, Streptococcus sanguinis (LMG 14657), and Ssal, Streptococcus salivarius strain TOVE-R.

    Techniques Used: Activity Assay

    36) Product Images from "A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications"

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.02008

    (A) Coomassie Brilliant Blue staining of crude cell extracts from T. forsythia ATCC 43037 wild-type and glycosyltransferase-deficient mutants after separation on a 7.5% SDS-PA gel. The S-layer glycoproteins (labeled TfsA and TfsB) are indicated and the downshifts resulting from glycan truncation can be seen in the mutants. S-layer glycoprotein bands were further processed for MS analyses. PageRuler Plus Prestained Protein Ladder (Thermo Fisher Scientific) was used as a protein molecular weight marker. (B) Western-blots probed with α-TfsA and α-TfsB antiserum for confirmation of the identity of S-layer glycoproteins. Glycoproteins from all glycosyltransferase-deficient mutants (Δ gtfSMILE ) experienced a downshift resulting from glycan truncation, whereas the reconstituted strains (denoted with +) regained wild-type migration, indicating the presence of the complete mature glycan, proving successful recombination. (C, i–vi) ESI-MS sum spectra of β-eliminated TfsB O -glycans from T. forsythia wild-type and mutants. The glycan structures of the signals corresponding to the largest mass (bold m / z values) are shown in SNFG representations ( Varki et al., 2015 ). O -Glycan signals detected for the respective mutants were assigned based on the m / z mass differences corresponding to the loss of individual sugar units and/or modifications.
    Figure Legend Snippet: (A) Coomassie Brilliant Blue staining of crude cell extracts from T. forsythia ATCC 43037 wild-type and glycosyltransferase-deficient mutants after separation on a 7.5% SDS-PA gel. The S-layer glycoproteins (labeled TfsA and TfsB) are indicated and the downshifts resulting from glycan truncation can be seen in the mutants. S-layer glycoprotein bands were further processed for MS analyses. PageRuler Plus Prestained Protein Ladder (Thermo Fisher Scientific) was used as a protein molecular weight marker. (B) Western-blots probed with α-TfsA and α-TfsB antiserum for confirmation of the identity of S-layer glycoproteins. Glycoproteins from all glycosyltransferase-deficient mutants (Δ gtfSMILE ) experienced a downshift resulting from glycan truncation, whereas the reconstituted strains (denoted with +) regained wild-type migration, indicating the presence of the complete mature glycan, proving successful recombination. (C, i–vi) ESI-MS sum spectra of β-eliminated TfsB O -glycans from T. forsythia wild-type and mutants. The glycan structures of the signals corresponding to the largest mass (bold m / z values) are shown in SNFG representations ( Varki et al., 2015 ). O -Glycan signals detected for the respective mutants were assigned based on the m / z mass differences corresponding to the loss of individual sugar units and/or modifications.

    Techniques Used: Staining, Labeling, Mass Spectrometry, Molecular Weight, Marker, Western Blot, Migration

    Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.
    Figure Legend Snippet: Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.

    Techniques Used: Sequencing

    ESI-MS sum spectra of β-eliminated TfsB O -glycans from T. forsythia ATCC 43037 methyltransferase knock-out mutants. The glycan structures of the signals corresponding to the largest mass (bold m / z values) are shown in SNFG representation ( Varki et al., 2015 ). Other O -glycan signals detected for the respective mutants were assigned based on the m / z mass differences corresponding to the loss of individual sugar units and/or modifications. The lack of methyl modifications is indicated by a red circle in the symbolic O -glycan structure representation.
    Figure Legend Snippet: ESI-MS sum spectra of β-eliminated TfsB O -glycans from T. forsythia ATCC 43037 methyltransferase knock-out mutants. The glycan structures of the signals corresponding to the largest mass (bold m / z values) are shown in SNFG representation ( Varki et al., 2015 ). Other O -glycan signals detected for the respective mutants were assigned based on the m / z mass differences corresponding to the loss of individual sugar units and/or modifications. The lack of methyl modifications is indicated by a red circle in the symbolic O -glycan structure representation.

    Techniques Used: Mass Spectrometry, Knock-Out

    (A) Scheme of the T. forsythia ATCC 43037 S-layer O -glycan structure. Monosaccharide symbols are shown according to the Symbol Nomenclature for Glycans (SNFG) ( Varki et al., 2015 ). Please note that the position of the branching Fuc remained unclear ( Posch et al., 2011 ) until it was determined in the course of this study to be on the reducing-end Gal. (B) Scheme of the 27-kb protein O -glycosylation gene cluster of T. forsythia ATCC 43037. Wzx (black), flippase; pseBCFHGI (green), CMP-Pse biosynthesis genes; gtfSMILE (blue), Gtf genes; mtfJOY (yellow), Mtf genes; asnB (putative asparagine synthetase B), wecC (UDP- N -acetyl- D -mannosamine dehydrogenase) and wecB (UDP- N- acetylglucosamine 2-epimerase) (purple); hypothetical proteins, HP (gray). Genes are not drawn to scale.
    Figure Legend Snippet: (A) Scheme of the T. forsythia ATCC 43037 S-layer O -glycan structure. Monosaccharide symbols are shown according to the Symbol Nomenclature for Glycans (SNFG) ( Varki et al., 2015 ). Please note that the position of the branching Fuc remained unclear ( Posch et al., 2011 ) until it was determined in the course of this study to be on the reducing-end Gal. (B) Scheme of the 27-kb protein O -glycosylation gene cluster of T. forsythia ATCC 43037. Wzx (black), flippase; pseBCFHGI (green), CMP-Pse biosynthesis genes; gtfSMILE (blue), Gtf genes; mtfJOY (yellow), Mtf genes; asnB (putative asparagine synthetase B), wecC (UDP- N -acetyl- D -mannosamine dehydrogenase) and wecB (UDP- N- acetylglucosamine 2-epimerase) (purple); hypothetical proteins, HP (gray). Genes are not drawn to scale.

    Techniques Used:

    Model for the biosynthesis of the species-specific portion of the T. forsythia ATCC 43037 O -glycan. Upon synthesis of the pentasaccharide core on an undP lipid carrier, the first carbohydrate residue of the species-specific glycan is a Fuc residue conferred by GtfE. The glycan is elongated with a Gal residue which is transferred by GtfL and methylated by MtfY. The assembly of the three sugar branch, consisting of a ManNAcA residue (transferred by GtfI), a ManNAcCONH 2 residue (GtfM), which is methylated by either MtfJ or MtfO, and a Pse5Am7Gra residue (transferred via GtfS), completes the synthesis of the decasaccharide.
    Figure Legend Snippet: Model for the biosynthesis of the species-specific portion of the T. forsythia ATCC 43037 O -glycan. Upon synthesis of the pentasaccharide core on an undP lipid carrier, the first carbohydrate residue of the species-specific glycan is a Fuc residue conferred by GtfE. The glycan is elongated with a Gal residue which is transferred by GtfL and methylated by MtfY. The assembly of the three sugar branch, consisting of a ManNAcA residue (transferred by GtfI), a ManNAcCONH 2 residue (GtfM), which is methylated by either MtfJ or MtfO, and a Pse5Am7Gra residue (transferred via GtfS), completes the synthesis of the decasaccharide.

    Techniques Used: Methylation

    37) Product Images from "Comparative genome characterization of the periodontal pathogen Tannerella forsythia"

    Article Title: Comparative genome characterization of the periodontal pathogen Tannerella forsythia

    Journal: BMC Genomics

    doi: 10.1186/s12864-020-6535-y

    Analysis of codon usage for ATCC 43037 (left panel) and BU063 (right panel). The continuous curves indicate the NC values to be expected for a given GC3s content in the absence of other factors shaping codon usage. Every dot represents a protein coding gene, dots not positioned near the curve therefore represent genes that display a considerable codon usage bias. GC3s: G + C content at synonymous positions, NC: effective number of codons used within the sequence of a gene
    Figure Legend Snippet: Analysis of codon usage for ATCC 43037 (left panel) and BU063 (right panel). The continuous curves indicate the NC values to be expected for a given GC3s content in the absence of other factors shaping codon usage. Every dot represents a protein coding gene, dots not positioned near the curve therefore represent genes that display a considerable codon usage bias. GC3s: G + C content at synonymous positions, NC: effective number of codons used within the sequence of a gene

    Techniques Used: Sequencing

    Multiple whole genome alignment of eight T. forsythia strains. Each coloured block represents a genomic region that aligned to a region in at least one other genome, plotted in the same colour, to which it was predicted to be homologous based on sequence similarity. Blocks above the centre line indicate forward orientation; blocks below the line indicate reverse orientation relative to strain 92A2. A histogram within each block shows the average similarity of a region to its counterparts in the other genomes. Red vertical lines indicate contig boundaries. Strain ATCC 43037 displayed two translocations compared to strain 92A2 with lengths of approximately 500 kbp (blue and yellow blocks at the right end of 92A2 and in the centre of ATCC) and 30 kbp (pink block at approx. 1.25 Mbp in 92A2 and at approx. 2.7 Mbp in ATCC), respectively. Previously described large-scale inversions in strain KS16 could be confirmed (reverted blocks in the left half of the alignment)
    Figure Legend Snippet: Multiple whole genome alignment of eight T. forsythia strains. Each coloured block represents a genomic region that aligned to a region in at least one other genome, plotted in the same colour, to which it was predicted to be homologous based on sequence similarity. Blocks above the centre line indicate forward orientation; blocks below the line indicate reverse orientation relative to strain 92A2. A histogram within each block shows the average similarity of a region to its counterparts in the other genomes. Red vertical lines indicate contig boundaries. Strain ATCC 43037 displayed two translocations compared to strain 92A2 with lengths of approximately 500 kbp (blue and yellow blocks at the right end of 92A2 and in the centre of ATCC) and 30 kbp (pink block at approx. 1.25 Mbp in 92A2 and at approx. 2.7 Mbp in ATCC), respectively. Previously described large-scale inversions in strain KS16 could be confirmed (reverted blocks in the left half of the alignment)

    Techniques Used: Blocking Assay, Sequencing

    Whole genome alignment between the six frame amino acid translations of both Tannerella sp. BU063 and the scaffolded and ordered ATCC 43037 assembly. Whereas the amino acid alignment reflects similarity with respect to gene content, the order of genes is not preserved
    Figure Legend Snippet: Whole genome alignment between the six frame amino acid translations of both Tannerella sp. BU063 and the scaffolded and ordered ATCC 43037 assembly. Whereas the amino acid alignment reflects similarity with respect to gene content, the order of genes is not preserved

    Techniques Used:

    38) Product Images from "Comparative genome characterization of the periodontal pathogen Tannerella forsythia"

    Article Title: Comparative genome characterization of the periodontal pathogen Tannerella forsythia

    Journal: BMC Genomics

    doi: 10.1186/s12864-020-6535-y

    Analysis of codon usage for ATCC 43037 (left panel) and BU063 (right panel). The continuous curves indicate the NC values to be expected for a given GC3s content in the absence of other factors shaping codon usage. Every dot represents a protein coding gene, dots not positioned near the curve therefore represent genes that display a considerable codon usage bias. GC3s: G + C content at synonymous positions, NC: effective number of codons used within the sequence of a gene
    Figure Legend Snippet: Analysis of codon usage for ATCC 43037 (left panel) and BU063 (right panel). The continuous curves indicate the NC values to be expected for a given GC3s content in the absence of other factors shaping codon usage. Every dot represents a protein coding gene, dots not positioned near the curve therefore represent genes that display a considerable codon usage bias. GC3s: G + C content at synonymous positions, NC: effective number of codons used within the sequence of a gene

    Techniques Used: Sequencing

    Multiple whole genome alignment of eight T. forsythia strains. Each coloured block represents a genomic region that aligned to a region in at least one other genome, plotted in the same colour, to which it was predicted to be homologous based on sequence similarity. Blocks above the centre line indicate forward orientation; blocks below the line indicate reverse orientation relative to strain 92A2. A histogram within each block shows the average similarity of a region to its counterparts in the other genomes. Red vertical lines indicate contig boundaries. Strain ATCC 43037 displayed two translocations compared to strain 92A2 with lengths of approximately 500 kbp (blue and yellow blocks at the right end of 92A2 and in the centre of ATCC) and 30 kbp (pink block at approx. 1.25 Mbp in 92A2 and at approx. 2.7 Mbp in ATCC), respectively. Previously described large-scale inversions in strain KS16 could be confirmed (reverted blocks in the left half of the alignment)
    Figure Legend Snippet: Multiple whole genome alignment of eight T. forsythia strains. Each coloured block represents a genomic region that aligned to a region in at least one other genome, plotted in the same colour, to which it was predicted to be homologous based on sequence similarity. Blocks above the centre line indicate forward orientation; blocks below the line indicate reverse orientation relative to strain 92A2. A histogram within each block shows the average similarity of a region to its counterparts in the other genomes. Red vertical lines indicate contig boundaries. Strain ATCC 43037 displayed two translocations compared to strain 92A2 with lengths of approximately 500 kbp (blue and yellow blocks at the right end of 92A2 and in the centre of ATCC) and 30 kbp (pink block at approx. 1.25 Mbp in 92A2 and at approx. 2.7 Mbp in ATCC), respectively. Previously described large-scale inversions in strain KS16 could be confirmed (reverted blocks in the left half of the alignment)

    Techniques Used: Blocking Assay, Sequencing

    Whole genome alignment between the six frame amino acid translations of both Tannerella sp. BU063 and the scaffolded and ordered ATCC 43037 assembly. Whereas the amino acid alignment reflects similarity with respect to gene content, the order of genes is not preserved
    Figure Legend Snippet: Whole genome alignment between the six frame amino acid translations of both Tannerella sp. BU063 and the scaffolded and ordered ATCC 43037 assembly. Whereas the amino acid alignment reflects similarity with respect to gene content, the order of genes is not preserved

    Techniques Used:

    39) Product Images from "Comparative genome characterization of the periodontal pathogen Tannerella forsythia"

    Article Title: Comparative genome characterization of the periodontal pathogen Tannerella forsythia

    Journal: BMC Genomics

    doi: 10.1186/s12864-020-6535-y

    Analysis of codon usage for ATCC 43037 (left panel) and BU063 (right panel). The continuous curves indicate the NC values to be expected for a given GC3s content in the absence of other factors shaping codon usage. Every dot represents a protein coding gene, dots not positioned near the curve therefore represent genes that display a considerable codon usage bias. GC3s: G + C content at synonymous positions, NC: effective number of codons used within the sequence of a gene
    Figure Legend Snippet: Analysis of codon usage for ATCC 43037 (left panel) and BU063 (right panel). The continuous curves indicate the NC values to be expected for a given GC3s content in the absence of other factors shaping codon usage. Every dot represents a protein coding gene, dots not positioned near the curve therefore represent genes that display a considerable codon usage bias. GC3s: G + C content at synonymous positions, NC: effective number of codons used within the sequence of a gene

    Techniques Used: Sequencing

    Multiple whole genome alignment of eight T. forsythia strains. Each coloured block represents a genomic region that aligned to a region in at least one other genome, plotted in the same colour, to which it was predicted to be homologous based on sequence similarity. Blocks above the centre line indicate forward orientation; blocks below the line indicate reverse orientation relative to strain 92A2. A histogram within each block shows the average similarity of a region to its counterparts in the other genomes. Red vertical lines indicate contig boundaries. Strain ATCC 43037 displayed two translocations compared to strain 92A2 with lengths of approximately 500 kbp (blue and yellow blocks at the right end of 92A2 and in the centre of ATCC) and 30 kbp (pink block at approx. 1.25 Mbp in 92A2 and at approx. 2.7 Mbp in ATCC), respectively. Previously described large-scale inversions in strain KS16 could be confirmed (reverted blocks in the left half of the alignment)
    Figure Legend Snippet: Multiple whole genome alignment of eight T. forsythia strains. Each coloured block represents a genomic region that aligned to a region in at least one other genome, plotted in the same colour, to which it was predicted to be homologous based on sequence similarity. Blocks above the centre line indicate forward orientation; blocks below the line indicate reverse orientation relative to strain 92A2. A histogram within each block shows the average similarity of a region to its counterparts in the other genomes. Red vertical lines indicate contig boundaries. Strain ATCC 43037 displayed two translocations compared to strain 92A2 with lengths of approximately 500 kbp (blue and yellow blocks at the right end of 92A2 and in the centre of ATCC) and 30 kbp (pink block at approx. 1.25 Mbp in 92A2 and at approx. 2.7 Mbp in ATCC), respectively. Previously described large-scale inversions in strain KS16 could be confirmed (reverted blocks in the left half of the alignment)

    Techniques Used: Blocking Assay, Sequencing

    Whole genome alignment between the six frame amino acid translations of both Tannerella sp. BU063 and the scaffolded and ordered ATCC 43037 assembly. Whereas the amino acid alignment reflects similarity with respect to gene content, the order of genes is not preserved
    Figure Legend Snippet: Whole genome alignment between the six frame amino acid translations of both Tannerella sp. BU063 and the scaffolded and ordered ATCC 43037 assembly. Whereas the amino acid alignment reflects similarity with respect to gene content, the order of genes is not preserved

    Techniques Used:

    40) Product Images from "Comparative genome characterization of the periodontal pathogen Tannerella forsythia"

    Article Title: Comparative genome characterization of the periodontal pathogen Tannerella forsythia

    Journal: BMC Genomics

    doi: 10.1186/s12864-020-6535-y

    Analysis of codon usage for ATCC 43037 (left panel) and BU063 (right panel). The continuous curves indicate the NC values to be expected for a given GC3s content in the absence of other factors shaping codon usage. Every dot represents a protein coding gene, dots not positioned near the curve therefore represent genes that display a considerable codon usage bias. GC3s: G + C content at synonymous positions, NC: effective number of codons used within the sequence of a gene
    Figure Legend Snippet: Analysis of codon usage for ATCC 43037 (left panel) and BU063 (right panel). The continuous curves indicate the NC values to be expected for a given GC3s content in the absence of other factors shaping codon usage. Every dot represents a protein coding gene, dots not positioned near the curve therefore represent genes that display a considerable codon usage bias. GC3s: G + C content at synonymous positions, NC: effective number of codons used within the sequence of a gene

    Techniques Used: Sequencing

    Multiple whole genome alignment of eight T. forsythia strains. Each coloured block represents a genomic region that aligned to a region in at least one other genome, plotted in the same colour, to which it was predicted to be homologous based on sequence similarity. Blocks above the centre line indicate forward orientation; blocks below the line indicate reverse orientation relative to strain 92A2. A histogram within each block shows the average similarity of a region to its counterparts in the other genomes. Red vertical lines indicate contig boundaries. Strain ATCC 43037 displayed two translocations compared to strain 92A2 with lengths of approximately 500 kbp (blue and yellow blocks at the right end of 92A2 and in the centre of ATCC) and 30 kbp (pink block at approx. 1.25 Mbp in 92A2 and at approx. 2.7 Mbp in ATCC), respectively. Previously described large-scale inversions in strain KS16 could be confirmed (reverted blocks in the left half of the alignment)
    Figure Legend Snippet: Multiple whole genome alignment of eight T. forsythia strains. Each coloured block represents a genomic region that aligned to a region in at least one other genome, plotted in the same colour, to which it was predicted to be homologous based on sequence similarity. Blocks above the centre line indicate forward orientation; blocks below the line indicate reverse orientation relative to strain 92A2. A histogram within each block shows the average similarity of a region to its counterparts in the other genomes. Red vertical lines indicate contig boundaries. Strain ATCC 43037 displayed two translocations compared to strain 92A2 with lengths of approximately 500 kbp (blue and yellow blocks at the right end of 92A2 and in the centre of ATCC) and 30 kbp (pink block at approx. 1.25 Mbp in 92A2 and at approx. 2.7 Mbp in ATCC), respectively. Previously described large-scale inversions in strain KS16 could be confirmed (reverted blocks in the left half of the alignment)

    Techniques Used: Blocking Assay, Sequencing

    Whole genome alignment between the six frame amino acid translations of both Tannerella sp. BU063 and the scaffolded and ordered ATCC 43037 assembly. Whereas the amino acid alignment reflects similarity with respect to gene content, the order of genes is not preserved
    Figure Legend Snippet: Whole genome alignment between the six frame amino acid translations of both Tannerella sp. BU063 and the scaffolded and ordered ATCC 43037 assembly. Whereas the amino acid alignment reflects similarity with respect to gene content, the order of genes is not preserved

    Techniques Used:

    Related Articles

    Mutagenesis:

    Article Title: Characterization of a potential ABC-type bacteriocin exporter protein from Treponema denticola
    Article Snippet: .. Construction of a tepA2 mutant As TepA2 showed similarity to ImmA, a tepA2- deficient mutant of T. denticola ATCC 35405 was constructed by allelic exchange mutation to investigate the role of tepA2 . .. Briefly, two fragments flanking the tepA2 gene were amplified with primer pairs 718D/719U and 719D/720U (listed in Table ), respectively.

    Article Title: Composition and Localization of Treponema denticola Outer Membrane Complexes ▿
    Article Snippet: .. T. denticola ATCC 35405 ( ) and isogenic msp mutant strain MHE ( ) were grown in NOS broth medium as previously described ( , ), with erythromycin (Em) (40 μg ml−1 ) added as appropriate. .. Cultures were examined by dark-field microscopy for purity and typical strain morphology.

    Sequencing:

    Article Title: Characterization of a potential ABC-type bacteriocin exporter protein from Treponema denticola
    Article Snippet: .. Sequence homology-based screening The whole genome sequence of T. denticola ATCC 35405 in the Los Alamos oral pathogen database ( http://www.oralgen.org ) was screened for homologous sequences with the S. mutans bacteriocin immunity protein (ImmA/Bip) sequence [ ] using the protein blast program. .. The obtained homologous sequences were further compared against the database of National Center for Biotechnology Information (NCBI, http://blast.ncbi.nlm.nih.gov/Blast.cgi ).

    other:

    Article Title: The central region of the msp gene of Treponema denticola has sequence heterogeneity among clinical samples, obtained from patients with periodontitis
    Article Snippet: Based on these findings, the 17 samples were divided into 2 distinct groups, A (15 samples) and B (the remaining 2), whose msp nucleotide sequences were closely related to T. denticola ATCC 35405 and ATCC 33520, respectively.

    Construct:

    Article Title: Characterization of a potential ABC-type bacteriocin exporter protein from Treponema denticola
    Article Snippet: .. Construction of a tepA2 mutant As TepA2 showed similarity to ImmA, a tepA2- deficient mutant of T. denticola ATCC 35405 was constructed by allelic exchange mutation to investigate the role of tepA2 . .. Briefly, two fragments flanking the tepA2 gene were amplified with primer pairs 718D/719U and 719D/720U (listed in Table ), respectively.

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    ATCC t forsythia atcc 43037 type strain
    (A) Coomassie Brilliant Blue staining of crude cell extracts from T. forsythia <t>ATCC</t> 43037 wild-type and glycosyltransferase-deficient mutants after separation on a 7.5% SDS-PA gel. The S-layer glycoproteins (labeled TfsA and TfsB) are indicated and the downshifts resulting from glycan truncation can be seen in the mutants. S-layer glycoprotein bands were further processed for MS analyses. PageRuler Plus Prestained Protein Ladder (Thermo Fisher Scientific) was used as a protein molecular weight marker. (B) Western-blots probed with α-TfsA and α-TfsB antiserum for confirmation of the identity of S-layer glycoproteins. Glycoproteins from all glycosyltransferase-deficient mutants (Δ gtfSMILE ) experienced a downshift resulting from glycan truncation, whereas the reconstituted strains (denoted with +) regained wild-type migration, indicating the presence of the complete mature glycan, proving successful recombination. (C, i–vi) ESI-MS sum spectra of β-eliminated TfsB O -glycans from T. forsythia wild-type and mutants. The glycan structures of the signals corresponding to the largest mass (bold m / z values) are shown in SNFG representations ( Varki et al., 2015 ). O -Glycan signals detected for the respective mutants were assigned based on the m / z mass differences corresponding to the loss of individual sugar units and/or modifications.
    T Forsythia Atcc 43037 Type Strain, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC t forsythia atcc 43037 genome
    The percentage sequence coverage of modern and ancient known virulence factor genes; the T. forsythia 92A2 sequence was used as a reference, apart from leg when <t>ATCC</t> 43037 was used as a reference; and KLIKK protease genes when our in-home determined KLIKK protease locus was used as a reference [ 10 ]
    T Forsythia Atcc 43037 Genome, supplied by ATCC, used in various techniques. Bioz Stars score: 89/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    ATCC t forsythia kinase tf murk
    The <t>rTf_MurK</t> reaction product is MurNAc-6-phosphate (MurNAc-6P). The product of the rTf_MurK <t>kinase</t> reaction with the substrate MurNAc, MurNAc-6P (red) (A) was degraded by the specific MurNAc-6P etherase of <t>T.</t> <t>forsythia</t> (rTf_MurQ) yielding the reaction product GlcNAc-6P (blue) as followed by LC-MS (B) . Shown are the extracted ion chromatograms (EICs) in negative ion mode: (M-H) - = 372.072 m/z for MurNAc-6P (red) at a retention time of 22 min and (M-H) - = 300.059 m/z for GlcNAc-6P (blue) at a retention time of 10 min on a Gemini C-18 RP-column.
    T Forsythia Kinase Tf Murk, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    ATCC t forsythia wild type strains
    Dual fluorescence in situ hybridization staining of Tannerella <t>forsythia</t> and Campylobacter rectus for biofilms harboring ATCC 43037 <t>wild‐type</t> (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: <t>T.</t> forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)
    T Forsythia Wild Type Strains, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 91 stars, based on 1 article reviews
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    Image Search Results


    (A) Coomassie Brilliant Blue staining of crude cell extracts from T. forsythia ATCC 43037 wild-type and glycosyltransferase-deficient mutants after separation on a 7.5% SDS-PA gel. The S-layer glycoproteins (labeled TfsA and TfsB) are indicated and the downshifts resulting from glycan truncation can be seen in the mutants. S-layer glycoprotein bands were further processed for MS analyses. PageRuler Plus Prestained Protein Ladder (Thermo Fisher Scientific) was used as a protein molecular weight marker. (B) Western-blots probed with α-TfsA and α-TfsB antiserum for confirmation of the identity of S-layer glycoproteins. Glycoproteins from all glycosyltransferase-deficient mutants (Δ gtfSMILE ) experienced a downshift resulting from glycan truncation, whereas the reconstituted strains (denoted with +) regained wild-type migration, indicating the presence of the complete mature glycan, proving successful recombination. (C, i–vi) ESI-MS sum spectra of β-eliminated TfsB O -glycans from T. forsythia wild-type and mutants. The glycan structures of the signals corresponding to the largest mass (bold m / z values) are shown in SNFG representations ( Varki et al., 2015 ). O -Glycan signals detected for the respective mutants were assigned based on the m / z mass differences corresponding to the loss of individual sugar units and/or modifications.

    Journal: Frontiers in Microbiology

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    doi: 10.3389/fmicb.2018.02008

    Figure Lengend Snippet: (A) Coomassie Brilliant Blue staining of crude cell extracts from T. forsythia ATCC 43037 wild-type and glycosyltransferase-deficient mutants after separation on a 7.5% SDS-PA gel. The S-layer glycoproteins (labeled TfsA and TfsB) are indicated and the downshifts resulting from glycan truncation can be seen in the mutants. S-layer glycoprotein bands were further processed for MS analyses. PageRuler Plus Prestained Protein Ladder (Thermo Fisher Scientific) was used as a protein molecular weight marker. (B) Western-blots probed with α-TfsA and α-TfsB antiserum for confirmation of the identity of S-layer glycoproteins. Glycoproteins from all glycosyltransferase-deficient mutants (Δ gtfSMILE ) experienced a downshift resulting from glycan truncation, whereas the reconstituted strains (denoted with +) regained wild-type migration, indicating the presence of the complete mature glycan, proving successful recombination. (C, i–vi) ESI-MS sum spectra of β-eliminated TfsB O -glycans from T. forsythia wild-type and mutants. The glycan structures of the signals corresponding to the largest mass (bold m / z values) are shown in SNFG representations ( Varki et al., 2015 ). O -Glycan signals detected for the respective mutants were assigned based on the m / z mass differences corresponding to the loss of individual sugar units and/or modifications.

    Article Snippet: Specifically, emphasis was focused on (i) sequence comparison of the general protein O- glycosylation gene cluster of the T. forsythia ATCC 43037 type strain with what is found in other T. forsythia strains for which genome sequences were available in public databases; (ii) transcriptional analysis of the protein O- glycosylation gene cluster of T forsythia ATCC 43037; (iii) construction of defined T. forsythia mutants deficient in predicted Gtfs and Mtfs encoded in the gene cluster and subsequent analysis of the O- glycans by MS to delineate the roles of the individual enzymes in glycan biosynthesis; and (iv) dissecting an O- glycan structure–function relationship in the immune response of DCs upon stimulation with the T. forsythia wild-type versus defined glycosylation-deficient mutants.

    Techniques: Staining, Labeling, Mass Spectrometry, Molecular Weight, Marker, Western Blot, Migration

    Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.

    Journal: Frontiers in Microbiology

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    doi: 10.3389/fmicb.2018.02008

    Figure Lengend Snippet: Alignment of protein O- glycosylation gene clusters from different T. forsythia strains showing comparable sizes and gene organizations (drawn to scale). Genes showing sequence identity > 50% and sequence coverage > 50% between strains appear in the same color. The major difference in the analyzed strains are for genes synthesizing either CMP-Pse ( pseBCFHGI ; light green colors; strains ATCC 43037 and UB20) or CMP-Leg ( legBCHIGF, ptmE ; dark green colors; strains FDC 92A2, UB4, KS16, UB22). Genes encoding Gtfs ( gtfSMILE ; blue color), Mtfs ( mtfJOY ; yellow color) and carbohydrate modifying enzymes ( asnB, wecC, wecB ; gray color) show high sequence homology between analyzed strains. Genomes of all strains synthesizing CMP-Leg encode an additional putative Mtf gene ( mtfX ), which does not share sequence homology to other Mtfs located within the cluster. In strain UB22, mtfJ is not predicted and for strain 3313 only five out of seven genes needed for the synthesis of CMP-Leg are predicted confidently. Due to low homology, isolate Tannerella sp. HOT-286 (phylotype BU063) could not be aligned with the other T. forsythia strains; for that isolate, the genomic area between a wzx -like gene and the gtfE gene is shown for comparison. P, Pse transferase; L, Leg transferase; HP, hypothetical protein; the star symbol ( ∗ ) indicates a transposable element; genes written in bold letters were investigated in detail in course of this study.

    Article Snippet: Specifically, emphasis was focused on (i) sequence comparison of the general protein O- glycosylation gene cluster of the T. forsythia ATCC 43037 type strain with what is found in other T. forsythia strains for which genome sequences were available in public databases; (ii) transcriptional analysis of the protein O- glycosylation gene cluster of T forsythia ATCC 43037; (iii) construction of defined T. forsythia mutants deficient in predicted Gtfs and Mtfs encoded in the gene cluster and subsequent analysis of the O- glycans by MS to delineate the roles of the individual enzymes in glycan biosynthesis; and (iv) dissecting an O- glycan structure–function relationship in the immune response of DCs upon stimulation with the T. forsythia wild-type versus defined glycosylation-deficient mutants.

    Techniques: Sequencing

    ESI-MS sum spectra of β-eliminated TfsB O -glycans from T. forsythia ATCC 43037 methyltransferase knock-out mutants. The glycan structures of the signals corresponding to the largest mass (bold m / z values) are shown in SNFG representation ( Varki et al., 2015 ). Other O -glycan signals detected for the respective mutants were assigned based on the m / z mass differences corresponding to the loss of individual sugar units and/or modifications. The lack of methyl modifications is indicated by a red circle in the symbolic O -glycan structure representation.

    Journal: Frontiers in Microbiology

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    doi: 10.3389/fmicb.2018.02008

    Figure Lengend Snippet: ESI-MS sum spectra of β-eliminated TfsB O -glycans from T. forsythia ATCC 43037 methyltransferase knock-out mutants. The glycan structures of the signals corresponding to the largest mass (bold m / z values) are shown in SNFG representation ( Varki et al., 2015 ). Other O -glycan signals detected for the respective mutants were assigned based on the m / z mass differences corresponding to the loss of individual sugar units and/or modifications. The lack of methyl modifications is indicated by a red circle in the symbolic O -glycan structure representation.

    Article Snippet: Specifically, emphasis was focused on (i) sequence comparison of the general protein O- glycosylation gene cluster of the T. forsythia ATCC 43037 type strain with what is found in other T. forsythia strains for which genome sequences were available in public databases; (ii) transcriptional analysis of the protein O- glycosylation gene cluster of T forsythia ATCC 43037; (iii) construction of defined T. forsythia mutants deficient in predicted Gtfs and Mtfs encoded in the gene cluster and subsequent analysis of the O- glycans by MS to delineate the roles of the individual enzymes in glycan biosynthesis; and (iv) dissecting an O- glycan structure–function relationship in the immune response of DCs upon stimulation with the T. forsythia wild-type versus defined glycosylation-deficient mutants.

    Techniques: Mass Spectrometry, Knock-Out

    (A) Scheme of the T. forsythia ATCC 43037 S-layer O -glycan structure. Monosaccharide symbols are shown according to the Symbol Nomenclature for Glycans (SNFG) ( Varki et al., 2015 ). Please note that the position of the branching Fuc remained unclear ( Posch et al., 2011 ) until it was determined in the course of this study to be on the reducing-end Gal. (B) Scheme of the 27-kb protein O -glycosylation gene cluster of T. forsythia ATCC 43037. Wzx (black), flippase; pseBCFHGI (green), CMP-Pse biosynthesis genes; gtfSMILE (blue), Gtf genes; mtfJOY (yellow), Mtf genes; asnB (putative asparagine synthetase B), wecC (UDP- N -acetyl- D -mannosamine dehydrogenase) and wecB (UDP- N- acetylglucosamine 2-epimerase) (purple); hypothetical proteins, HP (gray). Genes are not drawn to scale.

    Journal: Frontiers in Microbiology

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    doi: 10.3389/fmicb.2018.02008

    Figure Lengend Snippet: (A) Scheme of the T. forsythia ATCC 43037 S-layer O -glycan structure. Monosaccharide symbols are shown according to the Symbol Nomenclature for Glycans (SNFG) ( Varki et al., 2015 ). Please note that the position of the branching Fuc remained unclear ( Posch et al., 2011 ) until it was determined in the course of this study to be on the reducing-end Gal. (B) Scheme of the 27-kb protein O -glycosylation gene cluster of T. forsythia ATCC 43037. Wzx (black), flippase; pseBCFHGI (green), CMP-Pse biosynthesis genes; gtfSMILE (blue), Gtf genes; mtfJOY (yellow), Mtf genes; asnB (putative asparagine synthetase B), wecC (UDP- N -acetyl- D -mannosamine dehydrogenase) and wecB (UDP- N- acetylglucosamine 2-epimerase) (purple); hypothetical proteins, HP (gray). Genes are not drawn to scale.

    Article Snippet: Specifically, emphasis was focused on (i) sequence comparison of the general protein O- glycosylation gene cluster of the T. forsythia ATCC 43037 type strain with what is found in other T. forsythia strains for which genome sequences were available in public databases; (ii) transcriptional analysis of the protein O- glycosylation gene cluster of T forsythia ATCC 43037; (iii) construction of defined T. forsythia mutants deficient in predicted Gtfs and Mtfs encoded in the gene cluster and subsequent analysis of the O- glycans by MS to delineate the roles of the individual enzymes in glycan biosynthesis; and (iv) dissecting an O- glycan structure–function relationship in the immune response of DCs upon stimulation with the T. forsythia wild-type versus defined glycosylation-deficient mutants.

    Techniques:

    Model for the biosynthesis of the species-specific portion of the T. forsythia ATCC 43037 O -glycan. Upon synthesis of the pentasaccharide core on an undP lipid carrier, the first carbohydrate residue of the species-specific glycan is a Fuc residue conferred by GtfE. The glycan is elongated with a Gal residue which is transferred by GtfL and methylated by MtfY. The assembly of the three sugar branch, consisting of a ManNAcA residue (transferred by GtfI), a ManNAcCONH 2 residue (GtfM), which is methylated by either MtfJ or MtfO, and a Pse5Am7Gra residue (transferred via GtfS), completes the synthesis of the decasaccharide.

    Journal: Frontiers in Microbiology

    Article Title: A General Protein O-Glycosylation Gene Cluster Encodes the Species-Specific Glycan of the Oral Pathogen Tannerella forsythia: O-Glycan Biosynthesis and Immunological Implications

    doi: 10.3389/fmicb.2018.02008

    Figure Lengend Snippet: Model for the biosynthesis of the species-specific portion of the T. forsythia ATCC 43037 O -glycan. Upon synthesis of the pentasaccharide core on an undP lipid carrier, the first carbohydrate residue of the species-specific glycan is a Fuc residue conferred by GtfE. The glycan is elongated with a Gal residue which is transferred by GtfL and methylated by MtfY. The assembly of the three sugar branch, consisting of a ManNAcA residue (transferred by GtfI), a ManNAcCONH 2 residue (GtfM), which is methylated by either MtfJ or MtfO, and a Pse5Am7Gra residue (transferred via GtfS), completes the synthesis of the decasaccharide.

    Article Snippet: Specifically, emphasis was focused on (i) sequence comparison of the general protein O- glycosylation gene cluster of the T. forsythia ATCC 43037 type strain with what is found in other T. forsythia strains for which genome sequences were available in public databases; (ii) transcriptional analysis of the protein O- glycosylation gene cluster of T forsythia ATCC 43037; (iii) construction of defined T. forsythia mutants deficient in predicted Gtfs and Mtfs encoded in the gene cluster and subsequent analysis of the O- glycans by MS to delineate the roles of the individual enzymes in glycan biosynthesis; and (iv) dissecting an O- glycan structure–function relationship in the immune response of DCs upon stimulation with the T. forsythia wild-type versus defined glycosylation-deficient mutants.

    Techniques: Methylation

    The percentage sequence coverage of modern and ancient known virulence factor genes; the T. forsythia 92A2 sequence was used as a reference, apart from leg when ATCC 43037 was used as a reference; and KLIKK protease genes when our in-home determined KLIKK protease locus was used as a reference [ 10 ]

    Journal: BMC Genomics

    Article Title: Analysis of oral microbiome from fossil human remains revealed the significant differences in virulence factors of modern and ancient Tannerella forsythia

    doi: 10.1186/s12864-020-06810-9

    Figure Lengend Snippet: The percentage sequence coverage of modern and ancient known virulence factor genes; the T. forsythia 92A2 sequence was used as a reference, apart from leg when ATCC 43037 was used as a reference; and KLIKK protease genes when our in-home determined KLIKK protease locus was used as a reference [ 10 ]

    Article Snippet: Sequencing of the T. forsythia ATCC 43037 genome, apart from bspA (BFO_RS14480 ), revealed five more genes (BFO_RS14345 (bspB ), BFO_RS08355 , BFO_RS14330 , BFO_RS14330 , and BFO_RS14330 ) encoding putative BspA-like proteins.

    Techniques: Sequencing

    DNA sequence comparison of the  T. forsythia  reference genome to the ancient  T. forsythia  genomes and to publicly available modern  T. forsythia  genomes. The two outermost rings depict the forward and reverse coding strands of the reference genome. The next 13 rings moving towards the inner part of the figure display regions of sequence similarity detected by BLAST comparison between the DNA of the reference genome and the DNA of the 13 compared  T. forsythia  genomes. The following genome order reflects the order of the circles starting from the outer part of the figure and moving towards the inner part: PCA0332, PCA0198, UB20, PCA0088, KS16, UB4, UB22, NSLJ, G12, 3313, NSLK, ATCC 43037, and 9610. Genes associated with  T. forsythia  virulence are labeled in the plot

    Journal: BMC Genomics

    Article Title: Analysis of oral microbiome from fossil human remains revealed the significant differences in virulence factors of modern and ancient Tannerella forsythia

    doi: 10.1186/s12864-020-06810-9

    Figure Lengend Snippet: DNA sequence comparison of the T. forsythia reference genome to the ancient T. forsythia genomes and to publicly available modern T. forsythia genomes. The two outermost rings depict the forward and reverse coding strands of the reference genome. The next 13 rings moving towards the inner part of the figure display regions of sequence similarity detected by BLAST comparison between the DNA of the reference genome and the DNA of the 13 compared T. forsythia genomes. The following genome order reflects the order of the circles starting from the outer part of the figure and moving towards the inner part: PCA0332, PCA0198, UB20, PCA0088, KS16, UB4, UB22, NSLJ, G12, 3313, NSLK, ATCC 43037, and 9610. Genes associated with T. forsythia virulence are labeled in the plot

    Article Snippet: Sequencing of the T. forsythia ATCC 43037 genome, apart from bspA (BFO_RS14480 ), revealed five more genes (BFO_RS14345 (bspB ), BFO_RS08355 , BFO_RS14330 , BFO_RS14330 , and BFO_RS14330 ) encoding putative BspA-like proteins.

    Techniques: Sequencing, Labeling

    The rTf_MurK reaction product is MurNAc-6-phosphate (MurNAc-6P). The product of the rTf_MurK kinase reaction with the substrate MurNAc, MurNAc-6P (red) (A) was degraded by the specific MurNAc-6P etherase of T. forsythia (rTf_MurQ) yielding the reaction product GlcNAc-6P (blue) as followed by LC-MS (B) . Shown are the extracted ion chromatograms (EICs) in negative ion mode: (M-H) - = 372.072 m/z for MurNAc-6P (red) at a retention time of 22 min and (M-H) - = 300.059 m/z for GlcNAc-6P (blue) at a retention time of 10 min on a Gemini C-18 RP-column.

    Journal: Frontiers in Microbiology

    Article Title: N-Acetylmuramic Acid (MurNAc) Auxotrophy of the Oral Pathogen Tannerella forsythia: Characterization of a MurNAc Kinase and Analysis of Its Role in Cell Wall Metabolism

    doi: 10.3389/fmicb.2018.00019

    Figure Lengend Snippet: The rTf_MurK reaction product is MurNAc-6-phosphate (MurNAc-6P). The product of the rTf_MurK kinase reaction with the substrate MurNAc, MurNAc-6P (red) (A) was degraded by the specific MurNAc-6P etherase of T. forsythia (rTf_MurQ) yielding the reaction product GlcNAc-6P (blue) as followed by LC-MS (B) . Shown are the extracted ion chromatograms (EICs) in negative ion mode: (M-H) - = 372.072 m/z for MurNAc-6P (red) at a retention time of 22 min and (M-H) - = 300.059 m/z for GlcNAc-6P (blue) at a retention time of 10 min on a Gemini C-18 RP-column.

    Article Snippet: In the present study, we biochemically characterized the T. forsythia kinase Tf_MurK of the Tf_murTKQ operon from the type strain ATCC 43037 revealing stringent specificity of the enzyme for MurNAc.

    Techniques: Liquid Chromatography with Mass Spectroscopy

    Dual fluorescence in situ hybridization staining of Tannerella forsythia and Campylobacter rectus for biofilms harboring ATCC 43037 wild‐type (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: T. forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)

    Journal: Molecular Oral Microbiology

    Article Title: Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

    doi: 10.1111/omi.12182

    Figure Lengend Snippet: Dual fluorescence in situ hybridization staining of Tannerella forsythia and Campylobacter rectus for biofilms harboring ATCC 43037 wild‐type (A), UB 4 wild‐type (B), and ATCC 43037 ∆tfs AB (C). Red/yellow: T. forsythia , cyan: C. rectus; green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Scale bars 20 μm (A, B) and 15 μm (C)

    Article Snippet: 3.1 Monospecies biofilm formation of T. forsythia wild‐type strains and mutants Based on the observations that deficiency in the protein O ‐glycan's terminal nonulosonic acid triggers a decrease in biofilm formation of T. forsythia ATCC 43037 ∆pseC and T. forsythia UB4 ∆legC on a mucin‐coated surface and that T. forsythia ATCC 43037 ∆wecC possessing an even more truncated O ‐glycan forms more biofilm on untreated plates, the biofilm formation capacity of all these strains was compared here in one microtiter plate assay, where the plates were coated with mucin to mimic the native situation on the tooth surface, and biofilm growth was quantified by OD600 measurement of biofilm cells and normalized to the corresponding total cell mass for each strain.

    Techniques: Fluorescence, In Situ Hybridization, Staining

    Comparison of colony‐forming unit ( CFU ) counting and quantitative polymerase chain reaction ( qPCR ) for Tannerella forsythia wild‐type strains and mutants in the subgingival “Zurich biofilm”. Total bacteria for 10‐species biofilms with different T. forsythia strains and mutants enumerated by CFU counts (red boxes) and qPCR (blue boxes) for three independent experiments with three technical replicates, each, are shown (Whiskers boxplots 5th to 95th centile)

    Journal: Molecular Oral Microbiology

    Article Title: Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

    doi: 10.1111/omi.12182

    Figure Lengend Snippet: Comparison of colony‐forming unit ( CFU ) counting and quantitative polymerase chain reaction ( qPCR ) for Tannerella forsythia wild‐type strains and mutants in the subgingival “Zurich biofilm”. Total bacteria for 10‐species biofilms with different T. forsythia strains and mutants enumerated by CFU counts (red boxes) and qPCR (blue boxes) for three independent experiments with three technical replicates, each, are shown (Whiskers boxplots 5th to 95th centile)

    Article Snippet: 3.1 Monospecies biofilm formation of T. forsythia wild‐type strains and mutants Based on the observations that deficiency in the protein O ‐glycan's terminal nonulosonic acid triggers a decrease in biofilm formation of T. forsythia ATCC 43037 ∆pseC and T. forsythia UB4 ∆legC on a mucin‐coated surface and that T. forsythia ATCC 43037 ∆wecC possessing an even more truncated O ‐glycan forms more biofilm on untreated plates, the biofilm formation capacity of all these strains was compared here in one microtiter plate assay, where the plates were coated with mucin to mimic the native situation on the tooth surface, and biofilm growth was quantified by OD600 measurement of biofilm cells and normalized to the corresponding total cell mass for each strain.

    Techniques: Real-time Polymerase Chain Reaction

    Comparison of 10‐species biofilms with two Tannerella forsythia wild‐type strains. (A) Whiskers boxplots (5th to 95th centile) show bacterial numbers determined by quantitative real‐time PCR from three independent experiments. Asterisk (*) indicates a statistically significant difference ( P ≤.05) between groups. The two groups represent biofilms with either T. forsythia ATCC 43037 wild‐type or T. forsythia UB 4 wild‐type. (B, C) Fluorescence in situ hybridization stainings of fixed biofilms showing the localization of ATCC 43037 wild‐type (B) and UB 4 wild‐type (C). Red/yellow: T. forsythia; cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Here a representative area for one disk each is shown with a top view in the left panel and a side view with the biofilm–disk interface directed towards the top view; scale bars 5 μm (B) and 10 μm (C)

    Journal: Molecular Oral Microbiology

    Article Title: Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

    doi: 10.1111/omi.12182

    Figure Lengend Snippet: Comparison of 10‐species biofilms with two Tannerella forsythia wild‐type strains. (A) Whiskers boxplots (5th to 95th centile) show bacterial numbers determined by quantitative real‐time PCR from three independent experiments. Asterisk (*) indicates a statistically significant difference ( P ≤.05) between groups. The two groups represent biofilms with either T. forsythia ATCC 43037 wild‐type or T. forsythia UB 4 wild‐type. (B, C) Fluorescence in situ hybridization stainings of fixed biofilms showing the localization of ATCC 43037 wild‐type (B) and UB 4 wild‐type (C). Red/yellow: T. forsythia; cyan: Porphyromonas gingivalis , green: non‐hybridized cells ( DNA staining YoPro‐1+Sytox). Here a representative area for one disk each is shown with a top view in the left panel and a side view with the biofilm–disk interface directed towards the top view; scale bars 5 μm (B) and 10 μm (C)

    Article Snippet: 3.1 Monospecies biofilm formation of T. forsythia wild‐type strains and mutants Based on the observations that deficiency in the protein O ‐glycan's terminal nonulosonic acid triggers a decrease in biofilm formation of T. forsythia ATCC 43037 ∆pseC and T. forsythia UB4 ∆legC on a mucin‐coated surface and that T. forsythia ATCC 43037 ∆wecC possessing an even more truncated O ‐glycan forms more biofilm on untreated plates, the biofilm formation capacity of all these strains was compared here in one microtiter plate assay, where the plates were coated with mucin to mimic the native situation on the tooth surface, and biofilm growth was quantified by OD600 measurement of biofilm cells and normalized to the corresponding total cell mass for each strain.

    Techniques: Real-time Polymerase Chain Reaction, Fluorescence, In Situ Hybridization, Staining

    Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)

    Journal: Molecular Oral Microbiology

    Article Title: Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms. Behavior of two Tannerella forsythia strains and their cell surface mutants in multispecies oral biofilms

    doi: 10.1111/omi.12182

    Figure Lengend Snippet: Monospecies biofilm formation of Tannerella forsythia wild‐type and mutant strains. (A) Biofilm formation of T. forsythia ATCC 43037 wild‐type compared with its mutants ATCC 43037 Δ pseC , Δ wecC , Δ tfs AB and the complemented mutant Δ pseC comp . (B) Biofilm formation of T. forsythia UB 4 wild‐type compared with its mutant UB 4 Δ legC and the complemented mutant Δ legC comp . Mean values ± SD of four independent experiments with three replicates, each, are shown. Asterisks (**) indicate significant differences between samples as determined by the unpaired Student's t ‐test ( P ≤.01)

    Article Snippet: 3.1 Monospecies biofilm formation of T. forsythia wild‐type strains and mutants Based on the observations that deficiency in the protein O ‐glycan's terminal nonulosonic acid triggers a decrease in biofilm formation of T. forsythia ATCC 43037 ∆pseC and T. forsythia UB4 ∆legC on a mucin‐coated surface and that T. forsythia ATCC 43037 ∆wecC possessing an even more truncated O ‐glycan forms more biofilm on untreated plates, the biofilm formation capacity of all these strains was compared here in one microtiter plate assay, where the plates were coated with mucin to mimic the native situation on the tooth surface, and biofilm growth was quantified by OD600 measurement of biofilm cells and normalized to the corresponding total cell mass for each strain.

    Techniques: Mutagenesis