t denticola  (ATCC)


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    Structured Review

    ATCC t denticola
    TCT in the absence or presence of T. <t>denticola</t> ATCC 35405, MHE (Msp − ), or CKE (CTLP − ). Human plasma was incubated with T. denticola (1.2 × 10 9 cells/ml) suspended in PBS or with PBS alone (control) for 5 min before the addition
    T Denticola, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "The Chymotrypsin-Like Protease Complex of Treponema denticola ATCC 35405 Mediates Fibrinogen Adherence and Degradation "

    Article Title: The Chymotrypsin-Like Protease Complex of Treponema denticola ATCC 35405 Mediates Fibrinogen Adherence and Degradation

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00258-07

    TCT in the absence or presence of T. denticola ATCC 35405, MHE (Msp − ), or CKE (CTLP − ). Human plasma was incubated with T. denticola (1.2 × 10 9 cells/ml) suspended in PBS or with PBS alone (control) for 5 min before the addition
    Figure Legend Snippet: TCT in the absence or presence of T. denticola ATCC 35405, MHE (Msp − ), or CKE (CTLP − ). Human plasma was incubated with T. denticola (1.2 × 10 9 cells/ml) suspended in PBS or with PBS alone (control) for 5 min before the addition

    Techniques Used: Incubation

    Fibrinogen zymograms of outer membrane proteins from T. denticola ATCC 35405, MHE (Msp − ), and CKE (CTLP − ). Triton X-114 extracts were incubated for 5 min at 20°C (−) or 100°C (+) before being subjected to
    Figure Legend Snippet: Fibrinogen zymograms of outer membrane proteins from T. denticola ATCC 35405, MHE (Msp − ), and CKE (CTLP − ). Triton X-114 extracts were incubated for 5 min at 20°C (−) or 100°C (+) before being subjected to

    Techniques Used: Incubation

    Purification and enzymatic activity of major fibrinogen-binding protein from T. denticola ATCC 35405. Outer membrane proteins were extracted with 0.1% Triton X-114 solution and incubated with fibrinogen-linked Sepharose, and then tightly bound
    Figure Legend Snippet: Purification and enzymatic activity of major fibrinogen-binding protein from T. denticola ATCC 35405. Outer membrane proteins were extracted with 0.1% Triton X-114 solution and incubated with fibrinogen-linked Sepharose, and then tightly bound

    Techniques Used: Purification, Activity Assay, Binding Assay, Incubation

    Binding of fluid-phase or immobilized fibrinogen by T. denticola ATCC 35405 (⧫), MHE (Msp − ; ▴), or CKE (CTLP − ; ▪). (A and B) Treponema cells immobilized onto plastic wells in the absence (A) or presence (B) of 0.05
    Figure Legend Snippet: Binding of fluid-phase or immobilized fibrinogen by T. denticola ATCC 35405 (⧫), MHE (Msp − ; ▴), or CKE (CTLP − ; ▪). (A and B) Treponema cells immobilized onto plastic wells in the absence (A) or presence (B) of 0.05

    Techniques Used: Binding Assay

    Fibrinogen blot overlay of outer membrane protein extracts from T. denticola ATCC 35405, MHE, and CKE. Proteins were extracted with 1% Triton X-114 and incubated for 5 min at 20°C (−) or 100°C (+) before being subjected
    Figure Legend Snippet: Fibrinogen blot overlay of outer membrane protein extracts from T. denticola ATCC 35405, MHE, and CKE. Proteins were extracted with 1% Triton X-114 and incubated for 5 min at 20°C (−) or 100°C (+) before being subjected

    Techniques Used: Incubation

    Adherence of T. denticola ATCC 35405 cells to denatured fibrinogen or recombinant fibrinogen polypeptides. (A) Human fibrinogen (N) or purified recombinant Aα, Bβ, and γ chain polypeptides were heated for 5 min at 100°C
    Figure Legend Snippet: Adherence of T. denticola ATCC 35405 cells to denatured fibrinogen or recombinant fibrinogen polypeptides. (A) Human fibrinogen (N) or purified recombinant Aα, Bβ, and γ chain polypeptides were heated for 5 min at 100°C

    Techniques Used: Recombinant, Purification

    2) Product Images from "Porphyromonas gingivalis and Treponema denticola Exhibit Metabolic Symbioses"

    Article Title: Porphyromonas gingivalis and Treponema denticola Exhibit Metabolic Symbioses

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1003955

    Continuous culture of P. gingivalis and T. denticola mono- and co-cultures. Cell density of P. gingivalis and T. denticola mono- and co-cultures from three independent continuous cultures in OBGM with the dilution rate of 0.044 h −1 and mean generation time of 15.8 h as determined by measuring A 650 nm . The arrow shows the addition of P. gingivalis to a steady state T. denticola culture.
    Figure Legend Snippet: Continuous culture of P. gingivalis and T. denticola mono- and co-cultures. Cell density of P. gingivalis and T. denticola mono- and co-cultures from three independent continuous cultures in OBGM with the dilution rate of 0.044 h −1 and mean generation time of 15.8 h as determined by measuring A 650 nm . The arrow shows the addition of P. gingivalis to a steady state T. denticola culture.

    Techniques Used:

    Free glycine production during P. gingivalis growth. The difference in the amount of free glycine relative to that at t = 0 h as a function of P. gingivalis cell numbers in a) OBGM/PBS, b) OBGM and c) OBGM/ T. denticola conditioned medium. A regression line was fitted using a linear mixed modelling approach. The slope represents the amount of glycine produced/10 9 P. gingivalis cells.
    Figure Legend Snippet: Free glycine production during P. gingivalis growth. The difference in the amount of free glycine relative to that at t = 0 h as a function of P. gingivalis cell numbers in a) OBGM/PBS, b) OBGM and c) OBGM/ T. denticola conditioned medium. A regression line was fitted using a linear mixed modelling approach. The slope represents the amount of glycine produced/10 9 P. gingivalis cells.

    Techniques Used: Produced

    T. denticola growth and glycine. (a) The concentration of free glycine in T. denticola culture (black square; left axis). T. denticola growth curve in the same medium (black inverted triangle; right axis). Data points are the mean and standard deviation of three biological replicates. (b) Glycine (10 mM) was added to OBGM either before inoculation with T. denticola (black square, open arrow) or at 96 h after inoculation (black triangle, filled arrow) and bacterial growth was determined by A 650 nm measurement. T. denticola culture with no added glycine (white circle). Results are expressed as mean ± standard deviation obtained from eight replicates.
    Figure Legend Snippet: T. denticola growth and glycine. (a) The concentration of free glycine in T. denticola culture (black square; left axis). T. denticola growth curve in the same medium (black inverted triangle; right axis). Data points are the mean and standard deviation of three biological replicates. (b) Glycine (10 mM) was added to OBGM either before inoculation with T. denticola (black square, open arrow) or at 96 h after inoculation (black triangle, filled arrow) and bacterial growth was determined by A 650 nm measurement. T. denticola culture with no added glycine (white circle). Results are expressed as mean ± standard deviation obtained from eight replicates.

    Techniques Used: Concentration Assay, Standard Deviation

    Scanning electron micrograph of P. gingivalis and T. denticola grown in continuous culture. Co-culture was collected, fixed on a coverslip, dehydrated, covered with colloidal silver, gold-coated and imaged using a Philips XL30 field-emission scanning electron microscope. Electron micrographs showed that P. gingivalis and T. denticola coaggregated. T. denticola is a long helical shaped spirochete with an average length of 5 to 20 µm. P. gingivalis is a coccobacillus with an average diameter of 1 µm. Putative T. denticola outer sheath vesicles and/or P. gingivalis outer membrane vesicles are indicated by arrows along the length of T. denticola .
    Figure Legend Snippet: Scanning electron micrograph of P. gingivalis and T. denticola grown in continuous culture. Co-culture was collected, fixed on a coverslip, dehydrated, covered with colloidal silver, gold-coated and imaged using a Philips XL30 field-emission scanning electron microscope. Electron micrographs showed that P. gingivalis and T. denticola coaggregated. T. denticola is a long helical shaped spirochete with an average length of 5 to 20 µm. P. gingivalis is a coccobacillus with an average diameter of 1 µm. Putative T. denticola outer sheath vesicles and/or P. gingivalis outer membrane vesicles are indicated by arrows along the length of T. denticola .

    Techniques Used: Co-Culture Assay, Microscopy

    Proposed T. denticola glycine catabolic pathways. Glycine can be oxidized by the glycine cleavage system (1), producing NH 3 , CO 2 and CH 2 -THF. Glycine and CH 2 -THF can be condensed to form serine by serine hydroxymethyltransferase (2). Serine is deaminated to produce pyruvate by serine dehydratase (3). Lactate dehydrogenase (4) catalyzes the interconversion of pyruvate and lactate with concomitant interconversion of NADH and NAD + . Pyruvate can also be metabolized to acetate by pyruvate-ferredoxin oxidoreductase (5), phosphate acetyltransferase (6) and acetate kinase (7). Glycine can also be reduced to acetyl-P by the glycine reductase system (8).
    Figure Legend Snippet: Proposed T. denticola glycine catabolic pathways. Glycine can be oxidized by the glycine cleavage system (1), producing NH 3 , CO 2 and CH 2 -THF. Glycine and CH 2 -THF can be condensed to form serine by serine hydroxymethyltransferase (2). Serine is deaminated to produce pyruvate by serine dehydratase (3). Lactate dehydrogenase (4) catalyzes the interconversion of pyruvate and lactate with concomitant interconversion of NADH and NAD + . Pyruvate can also be metabolized to acetate by pyruvate-ferredoxin oxidoreductase (5), phosphate acetyltransferase (6) and acetate kinase (7). Glycine can also be reduced to acetyl-P by the glycine reductase system (8).

    Techniques Used:

    The extracellular products of T. denticola [U- 13 C]glycine fermentation. [U- 13 C]glycine (5.00 mM) was added to a 24 h T. denticola culture and aliquots were collected every 24 h, filtered and the identity and the quantity of the 13 C-labeled compounds was determined using NMR spectroscopy. black cross, [U- 13 C]glycine; black circle, [U- 13 C]acetate; black inverted triangle, dual or uniformly-labeled lactate; black triangle, [U- 13 C]bicarbonate; white circle, dual or uniformly-labeled alanine.
    Figure Legend Snippet: The extracellular products of T. denticola [U- 13 C]glycine fermentation. [U- 13 C]glycine (5.00 mM) was added to a 24 h T. denticola culture and aliquots were collected every 24 h, filtered and the identity and the quantity of the 13 C-labeled compounds was determined using NMR spectroscopy. black cross, [U- 13 C]glycine; black circle, [U- 13 C]acetate; black inverted triangle, dual or uniformly-labeled lactate; black triangle, [U- 13 C]bicarbonate; white circle, dual or uniformly-labeled alanine.

    Techniques Used: Labeling, Nuclear Magnetic Resonance, Spectroscopy

    P. gingivalis cell numbers and free glycine content in different cultures. a) The cell numbers of P. gingivalis in different media as determined by absorbance at 650 nm. b) The concentration of free glycine in different P. gingivalis cultures over time, as determined by GC-MS. Data shown are the average of three biological replicates. P. gingivalis grown in:- OBGM – black diamond; OBGM/PBS – black cross; OBGM/ T. denticola conditioned medium – white square. Uninoculated OBGM/ T. denticola conditioned medium – black square.
    Figure Legend Snippet: P. gingivalis cell numbers and free glycine content in different cultures. a) The cell numbers of P. gingivalis in different media as determined by absorbance at 650 nm. b) The concentration of free glycine in different P. gingivalis cultures over time, as determined by GC-MS. Data shown are the average of three biological replicates. P. gingivalis grown in:- OBGM – black diamond; OBGM/PBS – black cross; OBGM/ T. denticola conditioned medium – white square. Uninoculated OBGM/ T. denticola conditioned medium – black square.

    Techniques Used: Concentration Assay, Gas Chromatography, Mass Spectrometry

    3) Product Images from "Porphyromonas gingivalis and Treponema denticola Exhibit Metabolic Symbioses"

    Article Title: Porphyromonas gingivalis and Treponema denticola Exhibit Metabolic Symbioses

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1003955

    Continuous culture of P. gingivalis and T. denticola mono- and co-cultures. Cell density of P. gingivalis and T. denticola mono- and co-cultures from three independent continuous cultures in OBGM with the dilution rate of 0.044 h −1 and mean generation time of 15.8 h as determined by measuring A 650 nm . The arrow shows the addition of P. gingivalis to a steady state T. denticola culture.
    Figure Legend Snippet: Continuous culture of P. gingivalis and T. denticola mono- and co-cultures. Cell density of P. gingivalis and T. denticola mono- and co-cultures from three independent continuous cultures in OBGM with the dilution rate of 0.044 h −1 and mean generation time of 15.8 h as determined by measuring A 650 nm . The arrow shows the addition of P. gingivalis to a steady state T. denticola culture.

    Techniques Used:

    Free glycine production during P. gingivalis growth. The difference in the amount of free glycine relative to that at t = 0 h as a function of P. gingivalis cell numbers in a) OBGM/PBS, b) OBGM and c) OBGM/ T. denticola conditioned medium. A regression line was fitted using a linear mixed modelling approach. The slope represents the amount of glycine produced/10 9 P. gingivalis cells.
    Figure Legend Snippet: Free glycine production during P. gingivalis growth. The difference in the amount of free glycine relative to that at t = 0 h as a function of P. gingivalis cell numbers in a) OBGM/PBS, b) OBGM and c) OBGM/ T. denticola conditioned medium. A regression line was fitted using a linear mixed modelling approach. The slope represents the amount of glycine produced/10 9 P. gingivalis cells.

    Techniques Used: Produced

    T. denticola growth and glycine. (a) The concentration of free glycine in T. denticola culture (black square; left axis). T. denticola growth curve in the same medium (black inverted triangle; right axis). Data points are the mean and standard deviation of three biological replicates. (b) Glycine (10 mM) was added to OBGM either before inoculation with T. denticola (black square, open arrow) or at 96 h after inoculation (black triangle, filled arrow) and bacterial growth was determined by A 650 nm measurement. T. denticola culture with no added glycine (white circle). Results are expressed as mean ± standard deviation obtained from eight replicates.
    Figure Legend Snippet: T. denticola growth and glycine. (a) The concentration of free glycine in T. denticola culture (black square; left axis). T. denticola growth curve in the same medium (black inverted triangle; right axis). Data points are the mean and standard deviation of three biological replicates. (b) Glycine (10 mM) was added to OBGM either before inoculation with T. denticola (black square, open arrow) or at 96 h after inoculation (black triangle, filled arrow) and bacterial growth was determined by A 650 nm measurement. T. denticola culture with no added glycine (white circle). Results are expressed as mean ± standard deviation obtained from eight replicates.

    Techniques Used: Concentration Assay, Standard Deviation

    Scanning electron micrograph of P. gingivalis and T. denticola grown in continuous culture. Co-culture was collected, fixed on a coverslip, dehydrated, covered with colloidal silver, gold-coated and imaged using a Philips XL30 field-emission scanning electron microscope. Electron micrographs showed that P. gingivalis and T. denticola coaggregated. T. denticola is a long helical shaped spirochete with an average length of 5 to 20 µm. P. gingivalis is a coccobacillus with an average diameter of 1 µm. Putative T. denticola outer sheath vesicles and/or P. gingivalis outer membrane vesicles are indicated by arrows along the length of T. denticola .
    Figure Legend Snippet: Scanning electron micrograph of P. gingivalis and T. denticola grown in continuous culture. Co-culture was collected, fixed on a coverslip, dehydrated, covered with colloidal silver, gold-coated and imaged using a Philips XL30 field-emission scanning electron microscope. Electron micrographs showed that P. gingivalis and T. denticola coaggregated. T. denticola is a long helical shaped spirochete with an average length of 5 to 20 µm. P. gingivalis is a coccobacillus with an average diameter of 1 µm. Putative T. denticola outer sheath vesicles and/or P. gingivalis outer membrane vesicles are indicated by arrows along the length of T. denticola .

    Techniques Used: Co-Culture Assay, Microscopy

    Proposed T. denticola glycine catabolic pathways. Glycine can be oxidized by the glycine cleavage system (1), producing NH 3 , CO 2 and CH 2 -THF. Glycine and CH 2 -THF can be condensed to form serine by serine hydroxymethyltransferase (2). Serine is deaminated to produce pyruvate by serine dehydratase (3). Lactate dehydrogenase (4) catalyzes the interconversion of pyruvate and lactate with concomitant interconversion of NADH and NAD + . Pyruvate can also be metabolized to acetate by pyruvate-ferredoxin oxidoreductase (5), phosphate acetyltransferase (6) and acetate kinase (7). Glycine can also be reduced to acetyl-P by the glycine reductase system (8).
    Figure Legend Snippet: Proposed T. denticola glycine catabolic pathways. Glycine can be oxidized by the glycine cleavage system (1), producing NH 3 , CO 2 and CH 2 -THF. Glycine and CH 2 -THF can be condensed to form serine by serine hydroxymethyltransferase (2). Serine is deaminated to produce pyruvate by serine dehydratase (3). Lactate dehydrogenase (4) catalyzes the interconversion of pyruvate and lactate with concomitant interconversion of NADH and NAD + . Pyruvate can also be metabolized to acetate by pyruvate-ferredoxin oxidoreductase (5), phosphate acetyltransferase (6) and acetate kinase (7). Glycine can also be reduced to acetyl-P by the glycine reductase system (8).

    Techniques Used:

    The extracellular products of T. denticola [U- 13 C]glycine fermentation. [U- 13 C]glycine (5.00 mM) was added to a 24 h T. denticola culture and aliquots were collected every 24 h, filtered and the identity and the quantity of the 13 C-labeled compounds was determined using NMR spectroscopy. black cross, [U- 13 C]glycine; black circle, [U- 13 C]acetate; black inverted triangle, dual or uniformly-labeled lactate; black triangle, [U- 13 C]bicarbonate; white circle, dual or uniformly-labeled alanine.
    Figure Legend Snippet: The extracellular products of T. denticola [U- 13 C]glycine fermentation. [U- 13 C]glycine (5.00 mM) was added to a 24 h T. denticola culture and aliquots were collected every 24 h, filtered and the identity and the quantity of the 13 C-labeled compounds was determined using NMR spectroscopy. black cross, [U- 13 C]glycine; black circle, [U- 13 C]acetate; black inverted triangle, dual or uniformly-labeled lactate; black triangle, [U- 13 C]bicarbonate; white circle, dual or uniformly-labeled alanine.

    Techniques Used: Labeling, Nuclear Magnetic Resonance, Spectroscopy

    P. gingivalis cell numbers and free glycine content in different cultures. a) The cell numbers of P. gingivalis in different media as determined by absorbance at 650 nm. b) The concentration of free glycine in different P. gingivalis cultures over time, as determined by GC-MS. Data shown are the average of three biological replicates. P. gingivalis grown in:- OBGM – black diamond; OBGM/PBS – black cross; OBGM/ T. denticola conditioned medium – white square. Uninoculated OBGM/ T. denticola conditioned medium – black square.
    Figure Legend Snippet: P. gingivalis cell numbers and free glycine content in different cultures. a) The cell numbers of P. gingivalis in different media as determined by absorbance at 650 nm. b) The concentration of free glycine in different P. gingivalis cultures over time, as determined by GC-MS. Data shown are the average of three biological replicates. P. gingivalis grown in:- OBGM – black diamond; OBGM/PBS – black cross; OBGM/ T. denticola conditioned medium – white square. Uninoculated OBGM/ T. denticola conditioned medium – black square.

    Techniques Used: Concentration Assay, Gas Chromatography, Mass Spectrometry

    4) Product Images from "Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola"

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.
    Figure Legend Snippet: Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Techniques Used: Immuno-Electron Microscopy, Mutagenesis

    Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN
    Figure Legend Snippet: Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Techniques Used: Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Purification

    Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.
    Figure Legend Snippet: Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Techniques Used:

    Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.
    Figure Legend Snippet: Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Techniques Used: Activity Assay, Cell Culture

    Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.
    Figure Legend Snippet: Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Techniques Used: Sequencing

    Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard
    Figure Legend Snippet: Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Techniques Used: Incubation

    Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled
    Figure Legend Snippet: Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Techniques Used: Southern Blot, Mutagenesis, Labeling

    Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity
    Figure Legend Snippet: Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Techniques Used: Incubation, Activity Assay

    5) Product Images from "Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor"

    Article Title: Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor

    Journal: Molecular Microbiology

    doi: 10.1111/j.1365-2958.2008.06418.x

    The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay, Expressing, Construct, Mutagenesis, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation

    T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.
    Figure Legend Snippet: T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.

    Techniques Used: Expressing, Quantitative RT-PCR

    TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay

    TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.
    Figure Legend Snippet: Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.

    Techniques Used: Sequencing, Binding Assay

    Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.
    Figure Legend Snippet: Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.

    Techniques Used: Sequencing

    6) Product Images from "Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor"

    Article Title: Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor

    Journal: Molecular Microbiology

    doi: 10.1111/j.1365-2958.2008.06418.x

    The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay, Expressing, Construct, Mutagenesis, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation

    T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.
    Figure Legend Snippet: T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.

    Techniques Used: Expressing, Quantitative RT-PCR

    TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay

    TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.
    Figure Legend Snippet: Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.

    Techniques Used: Sequencing, Binding Assay

    Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.
    Figure Legend Snippet: Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.

    Techniques Used: Sequencing

    7) Product Images from "Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor"

    Article Title: Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor

    Journal: Molecular Microbiology

    doi: 10.1111/j.1365-2958.2008.06418.x

    The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay, Expressing, Construct, Mutagenesis, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation

    T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.
    Figure Legend Snippet: T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.

    Techniques Used: Expressing, Quantitative RT-PCR

    TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay

    TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.
    Figure Legend Snippet: Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.

    Techniques Used: Sequencing, Binding Assay

    Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.
    Figure Legend Snippet: Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.

    Techniques Used: Sequencing

    8) Product Images from "Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor"

    Article Title: Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor

    Journal: Molecular Microbiology

    doi: 10.1111/j.1365-2958.2008.06418.x

    The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay, Expressing, Construct, Mutagenesis, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation

    T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.
    Figure Legend Snippet: T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.

    Techniques Used: Expressing, Quantitative RT-PCR

    TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay

    TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.
    Figure Legend Snippet: Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.

    Techniques Used: Sequencing, Binding Assay

    Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.
    Figure Legend Snippet: Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.

    Techniques Used: Sequencing

    9) Product Images from "Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor"

    Article Title: Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor

    Journal: Molecular Microbiology

    doi: 10.1111/j.1365-2958.2008.06418.x

    The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay, Expressing, Construct, Mutagenesis, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation

    T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.
    Figure Legend Snippet: T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.

    Techniques Used: Expressing, Quantitative RT-PCR

    TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay

    TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.
    Figure Legend Snippet: Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.

    Techniques Used: Sequencing, Binding Assay

    Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.
    Figure Legend Snippet: Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.

    Techniques Used: Sequencing

    10) Product Images from "Binding Properties and Adhesion-Mediating Regions of the Major Sheath Protein of Treponema denticola ATCC 35405"

    Article Title: Binding Properties and Adhesion-Mediating Regions of the Major Sheath Protein of Treponema denticola ATCC 35405

    Journal: Infection and Immunity

    doi: 10.1128/IAI.73.5.2891-2898.2005

    Antibodies to rMsp inhibit T. denticola cell adhesion.
    Figure Legend Snippet: Antibodies to rMsp inhibit T. denticola cell adhesion.

    Techniques Used:

    Interactions of rMsp antibodies with T. denticola ATCC 35405 cells and of whole-cell antibodies with rMsp fragments. (Panel A) Reactivity in ELISA of antiserum (1:250 diluted) to T. denticola ATCC 35405 cells (W) or antisera (1:250 diluted) raised to
    Figure Legend Snippet: Interactions of rMsp antibodies with T. denticola ATCC 35405 cells and of whole-cell antibodies with rMsp fragments. (Panel A) Reactivity in ELISA of antiserum (1:250 diluted) to T. denticola ATCC 35405 cells (W) or antisera (1:250 diluted) raised to

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Reactivities of Msp antisera with T. denticola ATCC 35405 outer membrane protein extracts. (Panel A) SDS-PAGE gel of outer membrane proteins. (Panel B) Western blots reacted with antisera to rN-Msp (lane 1), rV-Msp (lane 2), or rC-Msp (lane 3). Molecular
    Figure Legend Snippet: Reactivities of Msp antisera with T. denticola ATCC 35405 outer membrane protein extracts. (Panel A) SDS-PAGE gel of outer membrane proteins. (Panel B) Western blots reacted with antisera to rN-Msp (lane 1), rV-Msp (lane 2), or rC-Msp (lane 3). Molecular

    Techniques Used: SDS Page, Western Blot

    Binding of rMsp to human fibronectin and to 30-kDa N-terminal fragment of human fibronectin. (Panel A) Western blot overlay of trypsin-derived fragments of human plasma fibronectin reacted with rMsp (lane 1) or with biotinylated T. denticola ATCC 35405
    Figure Legend Snippet: Binding of rMsp to human fibronectin and to 30-kDa N-terminal fragment of human fibronectin. (Panel A) Western blot overlay of trypsin-derived fragments of human plasma fibronectin reacted with rMsp (lane 1) or with biotinylated T. denticola ATCC 35405

    Techniques Used: Binding Assay, Western Blot, Derivative Assay

    Diagrammatic representation of the Msp sequences from T. denticola ATCC 35520 (A), strain ATCC 35405 (B), and of recombinant Msp polypeptides rMsp (530 aa residues), rN-Msp (189 aa residues), rV-Msp (57 aa residues), and rN-Msp (272 aa residues), with
    Figure Legend Snippet: Diagrammatic representation of the Msp sequences from T. denticola ATCC 35520 (A), strain ATCC 35405 (B), and of recombinant Msp polypeptides rMsp (530 aa residues), rN-Msp (189 aa residues), rV-Msp (57 aa residues), and rN-Msp (272 aa residues), with

    Techniques Used: Recombinant

    Effect of exogenously added rMsp on adhesion of Treponema strains to immobilized human fibronectin or 30-kDa fibronectin fragment. (Panel A) Adhesion levels of T. denticola ATCC 35405 cells to 0.1 μg fibronectin (filled column) or 30-kDa fibronectin
    Figure Legend Snippet: Effect of exogenously added rMsp on adhesion of Treponema strains to immobilized human fibronectin or 30-kDa fibronectin fragment. (Panel A) Adhesion levels of T. denticola ATCC 35405 cells to 0.1 μg fibronectin (filled column) or 30-kDa fibronectin

    Techniques Used:

    11) Product Images from "Molecular characterization of Treponema denticola infection-induced bone and soft tissue transcriptional profiles"

    Article Title: Molecular characterization of Treponema denticola infection-induced bone and soft tissue transcriptional profiles

    Journal:

    doi: 10.1111/j.2041-1014.2010.00575.x

    Cell Cycle pathway containing Treponema denticola differentially regulated genes in calvarial soft tissue compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes and Genomes nomenclature.
    Figure Legend Snippet: Cell Cycle pathway containing Treponema denticola differentially regulated genes in calvarial soft tissue compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes and Genomes nomenclature.

    Techniques Used: Infection

    Leukocyte transendothelial migration pathway containing Treponema denticola differentially regulated genes in calvarial bone compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes
    Figure Legend Snippet: Leukocyte transendothelial migration pathway containing Treponema denticola differentially regulated genes in calvarial bone compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes

    Techniques Used: Migration, Infection

    Leukocyte transendothelial migration pathway containing Treponema denticola differentially regulated genes in calvarial soft tissue compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia
    Figure Legend Snippet: Leukocyte transendothelial migration pathway containing Treponema denticola differentially regulated genes in calvarial soft tissue compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia

    Techniques Used: Migration, Infection

    Cell cycle pathway containing Treponema denticola differentially regulated genes in calvarial bone compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes and Genomes nomenclature.
    Figure Legend Snippet: Cell cycle pathway containing Treponema denticola differentially regulated genes in calvarial bone compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes and Genomes nomenclature.

    Techniques Used: Infection

    Effects of Treponema denticola local injection on mouse calvaria. Live T. denticola bacteria (1.5 × 10 9 ) were injected once daily for 3 days into the subcutaneous tissues overlying the calvaria of mice. All photomicrographs of slides stained with
    Figure Legend Snippet: Effects of Treponema denticola local injection on mouse calvaria. Live T. denticola bacteria (1.5 × 10 9 ) were injected once daily for 3 days into the subcutaneous tissues overlying the calvaria of mice. All photomicrographs of slides stained with

    Techniques Used: Injection, Mouse Assay, Staining

    Cell adhesion (immune system) molecules pathway containing Treponema denticola differentially regulated genes in calvarial bone (A) and tissue (B) compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto
    Figure Legend Snippet: Cell adhesion (immune system) molecules pathway containing Treponema denticola differentially regulated genes in calvarial bone (A) and tissue (B) compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto

    Techniques Used: Infection

    12) Product Images from "Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola"

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.
    Figure Legend Snippet: Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Techniques Used: Immuno-Electron Microscopy, Mutagenesis

    Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN
    Figure Legend Snippet: Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Techniques Used: Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Purification

    Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.
    Figure Legend Snippet: Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Techniques Used:

    Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.
    Figure Legend Snippet: Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Techniques Used: Activity Assay, Cell Culture

    Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.
    Figure Legend Snippet: Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Techniques Used: Sequencing

    Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard
    Figure Legend Snippet: Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Techniques Used: Incubation

    Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled
    Figure Legend Snippet: Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Techniques Used: Southern Blot, Mutagenesis, Labeling

    Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity
    Figure Legend Snippet: Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Techniques Used: Incubation, Activity Assay

    13) Product Images from "Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola"

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.
    Figure Legend Snippet: Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Techniques Used: Immuno-Electron Microscopy, Mutagenesis

    Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN
    Figure Legend Snippet: Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Techniques Used: Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Purification

    Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.
    Figure Legend Snippet: Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Techniques Used:

    Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.
    Figure Legend Snippet: Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Techniques Used: Activity Assay, Cell Culture

    Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.
    Figure Legend Snippet: Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Techniques Used: Sequencing

    Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard
    Figure Legend Snippet: Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Techniques Used: Incubation

    Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled
    Figure Legend Snippet: Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Techniques Used: Southern Blot, Mutagenesis, Labeling

    Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity
    Figure Legend Snippet: Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Techniques Used: Incubation, Activity Assay

    14) Product Images from "Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola"

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.
    Figure Legend Snippet: Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Techniques Used: Immuno-Electron Microscopy, Mutagenesis

    Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN
    Figure Legend Snippet: Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Techniques Used: Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Purification

    Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.
    Figure Legend Snippet: Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Techniques Used:

    Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.
    Figure Legend Snippet: Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Techniques Used: Activity Assay, Cell Culture

    Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.
    Figure Legend Snippet: Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Techniques Used: Sequencing

    Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard
    Figure Legend Snippet: Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Techniques Used: Incubation

    Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled
    Figure Legend Snippet: Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Techniques Used: Southern Blot, Mutagenesis, Labeling

    Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity
    Figure Legend Snippet: Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Techniques Used: Incubation, Activity Assay

    15) Product Images from "Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola"

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.
    Figure Legend Snippet: Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Techniques Used: Immuno-Electron Microscopy, Mutagenesis

    Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN
    Figure Legend Snippet: Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Techniques Used: Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Purification

    Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.
    Figure Legend Snippet: Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Techniques Used:

    Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.
    Figure Legend Snippet: Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Techniques Used: Activity Assay, Cell Culture

    Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.
    Figure Legend Snippet: Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Techniques Used: Sequencing

    Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard
    Figure Legend Snippet: Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Techniques Used: Incubation

    Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled
    Figure Legend Snippet: Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Techniques Used: Southern Blot, Mutagenesis, Labeling

    Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity
    Figure Legend Snippet: Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Techniques Used: Incubation, Activity Assay

    16) Product Images from "Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola"

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.
    Figure Legend Snippet: Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Techniques Used: Immuno-Electron Microscopy, Mutagenesis

    Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN
    Figure Legend Snippet: Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Techniques Used: Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Purification

    Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.
    Figure Legend Snippet: Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Techniques Used:

    Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.
    Figure Legend Snippet: Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Techniques Used: Activity Assay, Cell Culture

    Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.
    Figure Legend Snippet: Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Techniques Used: Sequencing

    Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard
    Figure Legend Snippet: Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Techniques Used: Incubation

    Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled
    Figure Legend Snippet: Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Techniques Used: Southern Blot, Mutagenesis, Labeling

    Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity
    Figure Legend Snippet: Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Techniques Used: Incubation, Activity Assay

    17) Product Images from "Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor"

    Article Title: Treponema denticola TroR is a manganese- and iron-dependent transcriptional repressor

    Journal: Molecular Microbiology

    doi: 10.1111/j.1365-2958.2008.06418.x

    The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: The TroR Td repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113 expressing pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) or (B) strain PAL113 expressing pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113 (expresses TroR Td ) were grown aerobically overnight in M9CG media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td deletion mutants exhibit variable repressor activity in E. coli . A. Physical maps of wild type and C-terminally truncated TroR Td proteins. TroR Td proteins are represented as grey lines and amino acid deletions are indicated. Names of the strains expressing the various TroR Td deletion constructs in the presence of pTDE100 ( T. denticola tro P/O- lacZ reporter) or pPAL100 ( T. pallidum tro P/O- lacZ reporter) are also indicated. DtxR secondary structure is indicated: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey, the proline-containing tether region (α7) is shown in black and SH3-like domain is shown in white. Full-length and mutant troR alleles were PCR-amplified from T. denticola ATCC 35405 chromosomal DNA and cloned into the NcoI and HindIII restriction sites of pBAD/HisA to facilitate the expression of TroR Td proteins. B. Immunoblot analysis of full-length and truncated TroR Td proteins expressed by: DEN113, DEN113A, DEN113B, DEN113C, DEN113D and DEN100. Effect of C-terminal deletion mutations on the expression of (C) pTDE100 or (D) pTPA100 reporter constructs. E. coli TOP10 cells harbouring both a TroR Td expression construct and a tro -P/O- lacZ reporter plasmid were grown aerobically overnight in LBL media. β-Galactosidase activity of the 12 h cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay, Expressing, Construct, Mutagenesis, Polymerase Chain Reaction, Amplification, Clone Assay, Plasmid Preparation

    T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.
    Figure Legend Snippet: T. denticola tro operon is negatively regulated by Mn 2+ and Fe 2+ . Expression of troA was analysed by qRT-PCR. RNA extracted from T. denticola grown in NOS-EC media supplemented with 5 μM Fe 2+ , Mn 2+ or Zn 2+ was quantified at (A) 24 h or (B) 48 h post inoculation using specific primers and probes with the Taqman system. Values have been normalized to the internal control, flaA . Fold changes are relative to spirochaetes grown in NOS-EC media lacking metal supplmentation (control). Results represent the means and standard deviations of three independent experiments performed in quadruplicate.

    Techniques Used: Expressing, Quantitative RT-PCR

    TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td Δ157−222 repressor responds to Mn 2+ and Fe 2+ in E. coli . (A) Strain DEN113A harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) or (B) strain PAL113A harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and pPJB113A (expresses TroR Td Δ157−222 ) were grown aerobically overnight in M9CG assay media supplemented with various divalent cations (5 μM) or in unsupplemented M9CG media (control). β-Galactosidase activity of the overnight cultures was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Activity Assay

    TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.
    Figure Legend Snippet: TroR Td expression negatively regulates both T. denticola and T. pallidum tro promoter activity. β-Galactosidase activity was measured from: (A) strains DEN100 and DEN113 harbouring pTDE100 ( T. denticola tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); (B) strains PAL100 and PAL113 harbouring pPAL100 ( T. pallidum tro P/O- lacZ reporter) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ); or (C) strains DEN103 and DEN103R harbouring pTDE103 ( T. denticola tro P/O- lacZ reporter with a disrupted operator region) and either pBAD/HisA (control) or pPJB113 (expresses TroR Td ). Cultures were grown aerobically for 12 h in LBL media and β-galactosidase activity was determined as described by Miller (1972) . Results represent the means and standard deviations of three independent experiments.

    Techniques Used: Expressing, Activity Assay

    Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.
    Figure Legend Snippet: Genetic organization of the T. pallidum and the T. denticola tro operons and sequence alignment of the T. pallidum and T. denticola tro P/O regions located immediately upstream of the troA ORFs. troA is predicted to encode a periplasmic binding protein; troB is predicted to encode the ATP-binding component, while troC and troD are predicted to encode the membrane components of ABC transporters; troR is homologous to a number of genes encoding Gram-positive iron-activated repressor proteins including DtxR and SirR; gpm encodes the glycolytic pathway enzyme, phosphoglycerate mutase. The σ 70 -like promoters (−10 and −35 sequences) are indicated in bold ( T. pallidum ) or underlined ( T. denitcola ). The 18 bp operator motifs are highlighted in grey. Sequence identity between the operators is indicated by vertical lines. Putative ribosome binding sites are in bold ( T. pallidum ) or underlined ( T. denitcola ) and indicated by SD. The troA start codons are indicated by boxes.

    Techniques Used: Sequencing, Binding Assay

    Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.
    Figure Legend Snippet: Primary sequence alignment of B. subtilis MntR (Bs MntR), C. diptheriae DtxR (Cd DtxR) and the T. pallidum (Tp) and T. denticola (Td) TroR proteins. The secondary structure of DtxR family metalloregulators is indicated above the sequence alignments: α-helices (α1, 2, 4, 5, 6) are shown in black, the DNA recognition helix (α3) is shown as a hatched line, β-strands (β1–2) are shown in grey and the proline-containing tether region (α7) is shown in black. The black boxes outline conserved metal co-ordination site residues in Bs MntR or Cd DtxR sequences. Putative TroR metal co-ordination site residues are highlighted in grey.

    Techniques Used: Sequencing

    18) Product Images from "Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola"

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.
    Figure Legend Snippet: Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Techniques Used: Immuno-Electron Microscopy, Mutagenesis

    Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN
    Figure Legend Snippet: Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Techniques Used: Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Purification

    Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.
    Figure Legend Snippet: Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Techniques Used:

    Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.
    Figure Legend Snippet: Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Techniques Used: Activity Assay, Cell Culture

    Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.
    Figure Legend Snippet: Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Techniques Used: Sequencing

    Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard
    Figure Legend Snippet: Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Techniques Used: Incubation

    Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled
    Figure Legend Snippet: Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Techniques Used: Southern Blot, Mutagenesis, Labeling

    Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity
    Figure Legend Snippet: Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Techniques Used: Incubation, Activity Assay

    19) Product Images from "Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola"

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.
    Figure Legend Snippet: Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Techniques Used: Immuno-Electron Microscopy, Mutagenesis

    Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN
    Figure Legend Snippet: Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Techniques Used: Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Purification

    Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.
    Figure Legend Snippet: Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Techniques Used:

    Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.
    Figure Legend Snippet: Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Techniques Used: Activity Assay, Cell Culture

    Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.
    Figure Legend Snippet: Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Techniques Used: Sequencing

    Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard
    Figure Legend Snippet: Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Techniques Used: Incubation

    Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled
    Figure Legend Snippet: Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Techniques Used: Southern Blot, Mutagenesis, Labeling

    Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity
    Figure Legend Snippet: Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Techniques Used: Incubation, Activity Assay

    20) Product Images from "Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola"

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    Journal: Infection and Immunity

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.
    Figure Legend Snippet: Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Techniques Used: Immuno-Electron Microscopy, Mutagenesis

    Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN
    Figure Legend Snippet: Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Techniques Used: Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Purification

    Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.
    Figure Legend Snippet: Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Techniques Used:

    Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.
    Figure Legend Snippet: Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Techniques Used: Activity Assay, Cell Culture

    Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.
    Figure Legend Snippet: Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Techniques Used: Sequencing

    Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard
    Figure Legend Snippet: Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Techniques Used: Incubation

    Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled
    Figure Legend Snippet: Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Techniques Used: Southern Blot, Mutagenesis, Labeling

    Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity
    Figure Legend Snippet: Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Techniques Used: Incubation, Activity Assay

    Related Articles

    Electroporation:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The resulting plasmid, pCRlrrA, was cut with AccI within the lrrA fragment, and an ermF-ermAM cassette from pVA2198 was inserted into the AccI site of pCRlrrA to construct plasmid plrrAerm. .. Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ). .. The erythromycin-resistant transformants were isolated and inoculated into TYGVS medium containing erythromycin (40 μg/ml).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA gene-defective mutant of T. denticola ATCC 35405 was isolated by a strategy commonly used for construction of T. denticola mutants in this laboratory ( ). .. The resulting plasmid, pCRlrrA, was cut with AccI within the lrrA fragment, and an ermF-ermAM cassette from pVA2198 was inserted into the AccI site of pCRlrrA to construct plasmid plrrAerm.

    Amplification:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The amplified fragment was cloned into the pCR2.1 vector (Invitrogen Corp., Carlsbad, Calif.). .. Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Ampicillin was added to Luria broth (LB) (Becton Dickinson and Co.) as indicated. .. The lrrA gene of T. denticola ATCC 35405 was amplified with oligonucleotide primers plrrAf (5′-AATCTTTAAGGAGGTTTGAC-3′) and plrrAr (5′-TATTTTGGGAAACGGCAG-3′) on the basis of the genome database of T. denticola from the Institute for Genomic Research ( ). .. The PCR product was treated with Exo/SAP (Amersham Biosciences, Piscataway, N.J.) and sequenced to confirm that its sequence was identical to that in the genome database of T. denticola ATCC 35405.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA gene-defective mutant of T. denticola ATCC 35405 was isolated by a strategy commonly used for construction of T. denticola mutants in this laboratory ( ). .. A portion of the lrrA gene was amplified with oligonucleotide primers plrrAfm (5′-AATAGGCTACAAGTGTGC-3′) and plrrArm (5′-TATTTTGGGAAACGGCAG-3′).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The amplified fragment, to which SalI and BglII recognition sites were added (shown in lowercase letters), was inserted into the SalI and BglII sites of shuttle vector pKMR4PE. .. The resulting plasmid, pKMlrrA, was transformed into T. denticola ATCC 33520.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA expression transformant of T. denticola ATCC 33520 was constructed with a T. denticola shuttle vector system ( ). .. The lrrA gene containing an upstream region including the Shine-Dalgarno sequence was amplified by PCR with primers pKMAf (5′-TTTTTgtcgacAATCTTTAAGGAGGTTTGACATG-3′) and pKMAr (5′-TTTTTagatctCTAGTTTTTCTTCCAGTATTTATC-3′).

    Southern Blot:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Plasmid plrrAerm was then linearized with EcoRV and electroporated into T. denticola ATCC 35405. .. Plasmid plrrAerm was then linearized with EcoRV and electroporated into T. denticola ATCC 35405.

    Clone Assay:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The amplified fragment was cloned into the pCR2.1 vector (Invitrogen Corp., Carlsbad, Calif.). .. Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Paragraph title: Cloning and sequencing of the lrrA gene of T. denticola . ... The lrrA gene of T. denticola ATCC 35405 was amplified with oligonucleotide primers plrrAf (5′-AATCTTTAAGGAGGTTTGAC-3′) and plrrAr (5′-TATTTTGGGAAACGGCAG-3′) on the basis of the genome database of T. denticola from the Institute for Genomic Research ( ).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA gene-defective mutant of T. denticola ATCC 35405 was isolated by a strategy commonly used for construction of T. denticola mutants in this laboratory ( ). .. A portion of the lrrA gene was amplified with oligonucleotide primers plrrAfm (5′-AATAGGCTACAAGTGTGC-3′) and plrrArm (5′-TATTTTGGGAAACGGCAG-3′).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: A 905-bp portion of the lrrA gene was cloned into vector pCR2.1, and a 2.4-kb ermF-ermAM cassette was inserted into the AccI site of the lrrA gene to construct plasmid plrrAerm. .. Plasmid plrrAerm was then linearized with EcoRV and electroporated into T. denticola ATCC 35405.

    Mutagenesis:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Paragraph title: Construction of the T. denticola lrrA mutant. ... Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: By contrast, mutant AK-1 was attenuated in coaggregation with Tannerella forsythensis (Fig. ). .. In addition, a reduction in coaggregation was likewise observed between T. denticola ATCC 35405 and the Tannerella forsythensis BspA mutant BFM-571 (Fig. ). .. Interestingly, strain ATCC 33520 appeared to aggregate with Tannerella forsythensis at a slower rate than strain ATCC 35405.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The PCR product was treated with Exo/SAP (Amersham Biosciences, Piscataway, N.J.) and sequenced to confirm that its sequence was identical to that in the genome database of T. denticola ATCC 35405. .. The lrrA gene-defective mutant of T. denticola ATCC 35405 was isolated by a strategy commonly used for construction of T. denticola mutants in this laboratory ( ). .. A portion of the lrrA gene was amplified with oligonucleotide primers plrrAfm (5′-AATAGGCTACAAGTGTGC-3′) and plrrArm (5′-TATTTTGGGAAACGGCAG-3′).

    Isolation:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA gene-defective mutant of T. denticola ATCC 35405 was isolated by a strategy commonly used for construction of T. denticola mutants in this laboratory ( ). .. Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The LRR sequence in T. denticola was identified in the genome database of T. denticola ATCC 35405 from TIGR with the BspA sequence. .. As a result, a gene encoding a highly conserved LRR sequence was found in the database in addition to several other potential LRR protein genes.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The PCR product was treated with Exo/SAP (Amersham Biosciences, Piscataway, N.J.) and sequenced to confirm that its sequence was identical to that in the genome database of T. denticola ATCC 35405. .. The lrrA gene-defective mutant of T. denticola ATCC 35405 was isolated by a strategy commonly used for construction of T. denticola mutants in this laboratory ( ). .. A portion of the lrrA gene was amplified with oligonucleotide primers plrrAfm (5′-AATAGGCTACAAGTGTGC-3′) and plrrArm (5′-TATTTTGGGAAACGGCAG-3′).

    Construct:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The resulting plasmid, pCRlrrA, was cut with AccI within the lrrA fragment, and an ermF-ermAM cassette from pVA2198 was inserted into the AccI site of pCRlrrA to construct plasmid plrrAerm. .. Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA gene-defective mutant of T. denticola ATCC 35405 was isolated by a strategy commonly used for construction of T. denticola mutants in this laboratory ( ). .. The amplified fragment was cloned into the pCR2.1 vector (Invitrogen Corp., Carlsbad, Calif.).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA expression transformant of T. denticola ATCC 33520 was constructed with a T. denticola shuttle vector system ( ). .. The resulting plasmid, pKMlrrA, was transformed into T. denticola ATCC 33520.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Digoxigenin labeling, hybridization, and detection were performed with a DIG DNA detection system (Roche Diagnostics Corporation, Indianapolis, Ind.) according to the manufacturer's protocol. .. The lrrA expression transformant of T. denticola ATCC 33520 was constructed with a T. denticola shuttle vector system ( ). .. The lrrA gene containing an upstream region including the Shine-Dalgarno sequence was amplified by PCR with primers pKMAf (5′-TTTTTgtcgacAATCTTTAAGGAGGTTTGACATG-3′) and pKMAr (5′-TTTTTagatctCTAGTTTTTCTTCCAGTATTTATC-3′).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: A 905-bp portion of the lrrA gene was cloned into vector pCR2.1, and a 2.4-kb ermF-ermAM cassette was inserted into the AccI site of the lrrA gene to construct plasmid plrrAerm. .. Plasmid plrrAerm was then linearized with EcoRV and electroporated into T. denticola ATCC 35405.

    Sequencing:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Paragraph title: Cloning and sequencing of the lrrA gene of T. denticola . ... The lrrA gene of T. denticola ATCC 35405 was amplified with oligonucleotide primers plrrAf (5′-AATCTTTAAGGAGGTTTGAC-3′) and plrrAr (5′-TATTTTGGGAAACGGCAG-3′) on the basis of the genome database of T. denticola from the Institute for Genomic Research ( ).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The nucleotide sequence of the lrrA gene has been deposited in the GenBank database and assigned accession number . .. The LRR sequence in T. denticola was identified in the genome database of T. denticola ATCC 35405 from TIGR with the BspA sequence. .. As a result, a gene encoding a highly conserved LRR sequence was found in the database in addition to several other potential LRR protein genes.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA gene containing an upstream region including the Shine-Dalgarno sequence was amplified by PCR with primers pKMAf (5′-TTTTTgtcgacAATCTTTAAGGAGGTTTGACATG-3′) and pKMAr (5′-TTTTTagatctCTAGTTTTTCTTCCAGTATTTATC-3′). .. The resulting plasmid, pKMlrrA, was transformed into T. denticola ATCC 33520.

    other:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: These results, taken together, suggested that the coaggregation between T. denticola and Tannerella forsythensis is mediated, in part, by interaction of the LrrA protein with the corresponding LRR protein, BspA, of Tannerella forsythensis .

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The association of LrrA with the surface of T. denticola is also consistent with the observation from this study that LrrA plays a role in interaction of this spirochete with its oral environment.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: To investigate the attachment of T. denticola to epithelial cells, attachment assays were performed with HEp-2 cell cultures.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: These results suggested that LrrA plays a role in the attachment of T. denticola to epithelial cells.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Neither T. denticola nor Tannerella forsythensis exhibited autoaggregation under these conditions.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: T. denticola has been reported to attach to epithelial cells and extracellular matrix components ( , , , - , , , ).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The present results regarding the attachment of T. denticola to HEp-2 cell tissue layers indicated that the LrrA protein plays a role in these interactions.

    Activity Assay:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Paragraph title: Coaggregation activity between T. denticola and heterologous bacteria. ... In addition, a reduction in coaggregation was likewise observed between T. denticola ATCC 35405 and the Tannerella forsythensis BspA mutant BFM-571 (Fig. ).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Attachment was calculated relative to that of strain ATCC 35405, which was adjusted to 100%. .. Coaggregation activity between T. denticola and heterologous bacteria was determined with cells washed and suspended in PBS. .. The percent coaggregation was calculated by the following equation: coaggregation = [(preincubation value [OD600 ] − sample value [OD600 ])/(preincubation value [OD600 ])] × 100.

    Infection:

    Article Title: Molecular characterization of Treponema denticola infection-induced bone and soft tissue transcriptional profiles
    Article Snippet: Further studies are required to identify specific T. denticola virulence factors (dentilisin, lipo-oligosaccharide) that alter host gene expression and measure the transcriptional response against a pathogenic strain compared with a mutant strain that lacks a virulence factor. .. Furthermore, the substantive impact of T. denticola in downregulation of host inflammatory/innate immune responses in this model supports the need for additional studies to explore the role of selected genes in the periodontal disease process, as well as the effect of T. denticola on host response patterns as a component of a polybacterial infection in periodontal disease. .. (82K, doc)

    Expressing:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Paragraph title: Expression of the lrrA gene in T. denticola . ... The resulting plasmid, pKMlrrA, was transformed into T. denticola ATCC 33520.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Digoxigenin labeling, hybridization, and detection were performed with a DIG DNA detection system (Roche Diagnostics Corporation, Indianapolis, Ind.) according to the manufacturer's protocol. .. The lrrA expression transformant of T. denticola ATCC 33520 was constructed with a T. denticola shuttle vector system ( ). .. The lrrA gene containing an upstream region including the Shine-Dalgarno sequence was amplified by PCR with primers pKMAf (5′-TTTTTgtcgacAATCTTTAAGGAGGTTTGACATG-3′) and pKMAr (5′-TTTTTagatctCTAGTTTTTCTTCCAGTATTTATC-3′).

    Protein Binding:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: LRRs have been reported to be generally useful as protein-binding motifs ( - , , ). .. In this study, an lrrA gene identified in the genome database of T. denticola ATCC 35405 was used to analyze the function of LRR proteins in T. denticola .

    Polymerase Chain Reaction:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA gene containing an upstream region including the Shine-Dalgarno sequence was amplified by PCR with primers pKMAf (5′-TTTTTgtcgacAATCTTTAAGGAGGTTTGACATG-3′) and pKMAr (5′-TTTTTagatctCTAGTTTTTCTTCCAGTATTTATC-3′). .. The resulting plasmid, pKMlrrA, was transformed into T. denticola ATCC 33520.

    Western Blot:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA expression transformant of T. denticola ATCC 33520 was constructed with a T. denticola shuttle vector system ( ). .. The resulting plasmid, pKMlrrA, was transformed into T. denticola ATCC 33520.

    Transformation Assay:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The amplified fragment, to which SalI and BglII recognition sites were added (shown in lowercase letters), was inserted into the SalI and BglII sites of shuttle vector pKMR4PE. .. The resulting plasmid, pKMlrrA, was transformed into T. denticola ATCC 33520. .. The transformant was examined for expression of the LrrA protein by Western blot analysis.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA expression transformant of T. denticola ATCC 33520 was constructed with a T. denticola shuttle vector system ( ). .. The amplified fragment, to which SalI and BglII recognition sites were added (shown in lowercase letters), was inserted into the SalI and BglII sites of shuttle vector pKMR4PE.

    Chemotaxis Assay:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Analysis of swarming indicated that the affected LrrA expression strains, mutant AK-1 and ATCC 33520/pKMlrrA, exhibited decreased swarming on TYGVS swarm plates, even though each appeared to be as motile as the parental strains in liquid cultures. .. This change may also attenuate tissue penetration, as demonstrated in the present study, because previous studies reported that the tissue penetration of oral epithelial cell layers by T. denticola is likely dependent upon motility and chemotaxis ( , ). .. The attenuated swarming by AK-1 suggested that the LrrA protein is involved in the swarming ability of strain ATCC 35405.

    Plasmid Preparation:

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The resulting plasmid, pCRlrrA, was cut with AccI within the lrrA fragment, and an ermF-ermAM cassette from pVA2198 was inserted into the AccI site of pCRlrrA to construct plasmid plrrAerm. .. Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ). .. The erythromycin-resistant transformants were isolated and inoculated into TYGVS medium containing erythromycin (40 μg/ml).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The lrrA gene-defective mutant of T. denticola ATCC 35405 was isolated by a strategy commonly used for construction of T. denticola mutants in this laboratory ( ). .. A portion of the lrrA gene was amplified with oligonucleotide primers plrrAfm (5′-AATAGGCTACAAGTGTGC-3′) and plrrArm (5′-TATTTTGGGAAACGGCAG-3′).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: The amplified fragment, to which SalI and BglII recognition sites were added (shown in lowercase letters), was inserted into the SalI and BglII sites of shuttle vector pKMR4PE. .. The resulting plasmid, pKMlrrA, was transformed into T. denticola ATCC 33520. .. The transformant was examined for expression of the LrrA protein by Western blot analysis.

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: Digoxigenin labeling, hybridization, and detection were performed with a DIG DNA detection system (Roche Diagnostics Corporation, Indianapolis, Ind.) according to the manufacturer's protocol. .. The lrrA expression transformant of T. denticola ATCC 33520 was constructed with a T. denticola shuttle vector system ( ). .. The lrrA gene containing an upstream region including the Shine-Dalgarno sequence was amplified by PCR with primers pKMAf (5′-TTTTTgtcgacAATCTTTAAGGAGGTTTGACATG-3′) and pKMAr (5′-TTTTTagatctCTAGTTTTTCTTCCAGTATTTATC-3′).

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola
    Article Snippet: A 905-bp portion of the lrrA gene was cloned into vector pCR2.1, and a 2.4-kb ermF-ermAM cassette was inserted into the AccI site of the lrrA gene to construct plasmid plrrAerm. .. Plasmid plrrAerm was then linearized with EcoRV and electroporated into T. denticola ATCC 35405. .. An insertion of the ermF-ermAM cassette into the lrrA gene on the chromosome by double crossover results in erythromycin-resistant (Ermr ) transformants (Fig. ).

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    ATCC t denticola
    Cell Cycle pathway containing Treponema <t>denticola</t> differentially regulated genes in calvarial soft tissue compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes and Genomes nomenclature.
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    Cell Cycle pathway containing Treponema denticola differentially regulated genes in calvarial soft tissue compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes and Genomes nomenclature.

    Journal:

    Article Title: Molecular characterization of Treponema denticola infection-induced bone and soft tissue transcriptional profiles

    doi: 10.1111/j.2041-1014.2010.00575.x

    Figure Lengend Snippet: Cell Cycle pathway containing Treponema denticola differentially regulated genes in calvarial soft tissue compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes and Genomes nomenclature.

    Article Snippet: Furthermore, the substantive impact of T. denticola in downregulation of host inflammatory/innate immune responses in this model supports the need for additional studies to explore the role of selected genes in the periodontal disease process, as well as the effect of T. denticola on host response patterns as a component of a polybacterial infection in periodontal disease.

    Techniques: Infection

    Leukocyte transendothelial migration pathway containing Treponema denticola differentially regulated genes in calvarial bone compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes

    Journal:

    Article Title: Molecular characterization of Treponema denticola infection-induced bone and soft tissue transcriptional profiles

    doi: 10.1111/j.2041-1014.2010.00575.x

    Figure Lengend Snippet: Leukocyte transendothelial migration pathway containing Treponema denticola differentially regulated genes in calvarial bone compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes

    Article Snippet: Furthermore, the substantive impact of T. denticola in downregulation of host inflammatory/innate immune responses in this model supports the need for additional studies to explore the role of selected genes in the periodontal disease process, as well as the effect of T. denticola on host response patterns as a component of a polybacterial infection in periodontal disease.

    Techniques: Migration, Infection

    Leukocyte transendothelial migration pathway containing Treponema denticola differentially regulated genes in calvarial soft tissue compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia

    Journal:

    Article Title: Molecular characterization of Treponema denticola infection-induced bone and soft tissue transcriptional profiles

    doi: 10.1111/j.2041-1014.2010.00575.x

    Figure Lengend Snippet: Leukocyte transendothelial migration pathway containing Treponema denticola differentially regulated genes in calvarial soft tissue compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia

    Article Snippet: Furthermore, the substantive impact of T. denticola in downregulation of host inflammatory/innate immune responses in this model supports the need for additional studies to explore the role of selected genes in the periodontal disease process, as well as the effect of T. denticola on host response patterns as a component of a polybacterial infection in periodontal disease.

    Techniques: Migration, Infection

    Cell cycle pathway containing Treponema denticola differentially regulated genes in calvarial bone compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes and Genomes nomenclature.

    Journal:

    Article Title: Molecular characterization of Treponema denticola infection-induced bone and soft tissue transcriptional profiles

    doi: 10.1111/j.2041-1014.2010.00575.x

    Figure Lengend Snippet: Cell cycle pathway containing Treponema denticola differentially regulated genes in calvarial bone compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto Encyclopedia of Genes and Genomes nomenclature.

    Article Snippet: Furthermore, the substantive impact of T. denticola in downregulation of host inflammatory/innate immune responses in this model supports the need for additional studies to explore the role of selected genes in the periodontal disease process, as well as the effect of T. denticola on host response patterns as a component of a polybacterial infection in periodontal disease.

    Techniques: Infection

    Effects of Treponema denticola local injection on mouse calvaria. Live T. denticola bacteria (1.5 × 10 9 ) were injected once daily for 3 days into the subcutaneous tissues overlying the calvaria of mice. All photomicrographs of slides stained with

    Journal:

    Article Title: Molecular characterization of Treponema denticola infection-induced bone and soft tissue transcriptional profiles

    doi: 10.1111/j.2041-1014.2010.00575.x

    Figure Lengend Snippet: Effects of Treponema denticola local injection on mouse calvaria. Live T. denticola bacteria (1.5 × 10 9 ) were injected once daily for 3 days into the subcutaneous tissues overlying the calvaria of mice. All photomicrographs of slides stained with

    Article Snippet: Furthermore, the substantive impact of T. denticola in downregulation of host inflammatory/innate immune responses in this model supports the need for additional studies to explore the role of selected genes in the periodontal disease process, as well as the effect of T. denticola on host response patterns as a component of a polybacterial infection in periodontal disease.

    Techniques: Injection, Mouse Assay, Staining

    Cell adhesion (immune system) molecules pathway containing Treponema denticola differentially regulated genes in calvarial bone (A) and tissue (B) compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto

    Journal:

    Article Title: Molecular characterization of Treponema denticola infection-induced bone and soft tissue transcriptional profiles

    doi: 10.1111/j.2041-1014.2010.00575.x

    Figure Lengend Snippet: Cell adhesion (immune system) molecules pathway containing Treponema denticola differentially regulated genes in calvarial bone (A) and tissue (B) compared with sham-infected controls at P ≤ 0.05, adapted from Pathway Express and using the Kyoto

    Article Snippet: Furthermore, the substantive impact of T. denticola in downregulation of host inflammatory/innate immune responses in this model supports the need for additional studies to explore the role of selected genes in the periodontal disease process, as well as the effect of T. denticola on host response patterns as a component of a polybacterial infection in periodontal disease.

    Techniques: Infection

    Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Journal: Infection and Immunity

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Figure Lengend Snippet: Immunoelectron microscopy of T. denticola ATCC 35405 (A) and LrrA-deficient mutant AK-1 (B) cells with anti-LrrA serum. The size of the gold particles is 10 nm.

    Article Snippet: Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Techniques: Immuno-Electron Microscopy, Mutagenesis

    Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Journal: Infection and Immunity

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Figure Lengend Snippet: Western blot analysis of expression of the LrrA protein in different T. denticola strains. Proteins were separated by SDS-12% PAGE and transferred onto a polyvinylidene difluoride membrane. Lanes: 1, molecular mass markers; 2, purified His-LrrAΔN

    Article Snippet: Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Techniques: Western Blot, Expressing, Polyacrylamide Gel Electrophoresis, Purification

    Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Journal: Infection and Immunity

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Figure Lengend Snippet: Coaggregation between T. denticola and heterologous bacteria. (A) Coaggregation between T. denticola and heterologous bacteria. The percent coaggregation was calculated as described in the text. The data are the averaged values from three separate experiments.

    Article Snippet: Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Techniques:

    Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Journal: Infection and Immunity

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Figure Lengend Snippet: Swarming activity of T. denticola. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA cultured in TYGVS medium swarm plates (containing 0.8% agarose) were monitored visually after 7 days.

    Article Snippet: Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Techniques: Activity Assay, Cell Culture

    Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Journal: Infection and Immunity

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Figure Lengend Snippet: Nucleotide and deduced amino acid sequences of the lrrA gene of T. denticola ATCC 35405. The amino acid sequences of the LRR regions are shown in boldface letters and underlined. The potential Shine-Dalgarno sequence (SD) and spirochetal lipobox are underlined.

    Article Snippet: Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Techniques: Sequencing

    Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Journal: Infection and Immunity

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Figure Lengend Snippet: Tissue penetration of T. denticola . Penetration rates were determined by counting cell numbers after 8 h of incubation of T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA with HEp-2 cell tissue layers. Values are means and standard

    Article Snippet: Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Techniques: Incubation

    Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Journal: Infection and Immunity

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Figure Lengend Snippet: Southern blot analysis of T. denticola ATCC 35405, the lrrA :: erm mutant AK-1, and strain ATCC 33520. Chromosomal DNAs from strain ATCC 33520 (lane 1), strain ATCC 35405 (lane 2), and mutant AK-1 (lane 3) were digested with PvuII. The digoxigenin-labeled

    Article Snippet: Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Techniques: Southern Blot, Mutagenesis, Labeling

    Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Journal: Infection and Immunity

    Article Title: Multiple Functions of the Leucine-Rich Repeat Protein LrrA of Treponema denticola

    doi: 10.1128/IAI.72.8.4619-4627.2004

    Figure Lengend Snippet: Attachment of T. denticola strains to HEp-2 cells. T. denticola strains ATCC 35405, AK-1, ATCC 33520, and ATCC 33520/pKMlrrA) were added to HEp-2 cell tissue layers and incubated for 1 h at 37°C. Attached cells were detected by chymotrypsin activity

    Article Snippet: Plasmid plrrAerm was then linearized following EcoRV digestion, and 10 μg of linearized plasmid was used to transform T. denticola ATCC 35405 by electroporation as described previously ( ).

    Techniques: Incubation, Activity Assay