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  • 99
    ATCC t denticola atcc 35405
    Construction of T. <t>denticola</t> mutants using ermF-ermB and ermB cassettes. The protease operon ( prcB-prcA-prtP ) and adjacent gene TDE0759 in wildtype parent strain <t>ATCC</t> 35405 and isogenic strains mutated in prcB (P0760 and CF548) and prcA (PNE and CF547) showing locations of ermF-ermB and ermB insertions. The symbol x- indicates gene disruption in prcA or prcB .
    T Denticola Atcc 35405, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 703 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    ATCC t denticola atcc 35404
    T. <t>denticola</t> proteases are not responsible for resistance to hβD-2. (A) T. denticola <t>ATCC</t> 35404 incubated in the presence or absence of 100 μM chymostatin (CHY) remained resistant to 10 μg of hβD-2 per ml over a 4-h period. (B) E. coli was incubated in the presence of 10 μg of hβD-2 per ml for 4 h, with or without equal numbers of T. denticola ATCC 35404. Data are means ± SEM from five (A) and three (B) experiments. SDS, sodium dodecyl sulfate.
    T Denticola Atcc 35404, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 138 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    ATCC t denticola atcc 33520
    Coaggregation and biofilm formation of T. <t>denticola</t> wild-type, Δ flgE , Δ motB with P. gingivalis W50. (A) Coaggregation of T. denticola wild-type and mutants with P. gingivalis W50. T. denticola <t>ATCC</t> 33520 and P. gingivalis cells at exponential growth phase were harvested, washed twice with coaggregation buffer and adjusted to A 650 of 0.5 with the coaggregation buffer. Equal volumes of T. denticola and P. gingivalis cell suspensions were combined. The height of bacterial aggregates was monitored for 7 h. The data are presented as means plus standard deviations ( N = 3). (B) Monospecies and dual-species static biofilms of T. denticola ATCC 33520 wild-type, Δ flgE , Δ motB with P. gingivalis W50. T. denticola and P. gingivalis cells were grown to exponential phase, diluted to A 650 of 0.15 and grown anaerobically for 5 days in a 12-well plate, either in a monospecies or dual-species culture. The resultant biofilm was stained with crystal violet and the total biomass determined spectrophotometrically. The data are presented as means and standard deviations ( N = 23–27) and were analyzed using a Kruskal-Wallis with Conover-Imam test. All values were significantly different ( p
    T Denticola Atcc 33520, supplied by ATCC, used in various techniques. Bioz Stars score: 97/100, based on 122 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    ATCC t denticola atcc 33521
    Coaggregation and biofilm formation of T. <t>denticola</t> wild-type, Δ flgE , Δ motB with P. gingivalis W50. (A) Coaggregation of T. denticola wild-type and mutants with P. gingivalis W50. T. denticola <t>ATCC</t> 33520 and P. gingivalis cells at exponential growth phase were harvested, washed twice with coaggregation buffer and adjusted to A 650 of 0.5 with the coaggregation buffer. Equal volumes of T. denticola and P. gingivalis cell suspensions were combined. The height of bacterial aggregates was monitored for 7 h. The data are presented as means plus standard deviations ( N = 3). (B) Monospecies and dual-species static biofilms of T. denticola ATCC 33520 wild-type, Δ flgE , Δ motB with P. gingivalis W50. T. denticola and P. gingivalis cells were grown to exponential phase, diluted to A 650 of 0.15 and grown anaerobically for 5 days in a 12-well plate, either in a monospecies or dual-species culture. The resultant biofilm was stained with crystal violet and the total biomass determined spectrophotometrically. The data are presented as means and standard deviations ( N = 23–27) and were analyzed using a Kruskal-Wallis with Conover-Imam test. All values were significantly different ( p
    T Denticola Atcc 33521, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    ATCC t denticola atcc 700768
    Coaggregation and biofilm formation of T. <t>denticola</t> wild-type, Δ flgE , Δ motB with P. gingivalis W50. (A) Coaggregation of T. denticola wild-type and mutants with P. gingivalis W50. T. denticola <t>ATCC</t> 33520 and P. gingivalis cells at exponential growth phase were harvested, washed twice with coaggregation buffer and adjusted to A 650 of 0.5 with the coaggregation buffer. Equal volumes of T. denticola and P. gingivalis cell suspensions were combined. The height of bacterial aggregates was monitored for 7 h. The data are presented as means plus standard deviations ( N = 3). (B) Monospecies and dual-species static biofilms of T. denticola ATCC 33520 wild-type, Δ flgE , Δ motB with P. gingivalis W50. T. denticola and P. gingivalis cells were grown to exponential phase, diluted to A 650 of 0.15 and grown anaerobically for 5 days in a 12-well plate, either in a monospecies or dual-species culture. The resultant biofilm was stained with crystal violet and the total biomass determined spectrophotometrically. The data are presented as means and standard deviations ( N = 23–27) and were analyzed using a Kruskal-Wallis with Conover-Imam test. All values were significantly different ( p
    T Denticola Atcc 700768, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Softberry Inc t denticola genome sequence
    Interaction of PrcB with PrtP. Arrows denote PrcB, PrtP, and rabbit IgG heavy chains (H). (A) Immunoblots probed with anti-PrtP antibodies and HisProbe reagent, as indicated. Lanes 1 and 2, T. <t>denticola</t> CF499; lanes 3 and 4, CF417; lanes 1 and 4, cell extracts; lanes 2 and 3, anti-PrtP immunoprecipitates from cell extracts. (B) T. denticola CF499 lysates were immunoprecipitated with polyclonal antibodies to PrtP, PrcA, FlaA, or Msp. Eluted proteins on duplicate blots were probed with anti-PrtP antibodies and HisProbe reagent, as indicated. (C) Immunoblot probed with anti-PrtP antibodies. Lanes 1 to 3, T. denticola CF499; lanes 4 to 6, T. denticola CF417; lanes 1 and 4, whole-cell extracts; lanes 2 and 5, anti-6×His monoclonal antibody immunoprecipitates from cell extracts; lanes 3 and 6, anti-PrtP antibody immunoprecipitates from cell extracts. (D) E. coli cells expressing PrtP, 6×His-PrcB, or both PrtP and 6×His-PrcB were analyzed directly and after Ni 2+ chromatography. Immunoblots of whole-cell extracts and Ni 2+ affinity column eluates were probed with anti-6×His monoclonal antibody or anti-PrtP polyclonal antibodies. Lanes: 1, E. coli /pCF411 (expresses PrtP); 2, E. coli /pCF415 (expresses PrcB-6×His); 3, E. coli /pCF416 (expresses PrcB-6×His + PrtP).
    T Denticola Genome Sequence, supplied by Softberry Inc, used in various techniques. Bioz Stars score: 85/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    ATCC t denticola atcc 33405
    Interaction of PrcB with PrtP. Arrows denote PrcB, PrtP, and rabbit IgG heavy chains (H). (A) Immunoblots probed with anti-PrtP antibodies and HisProbe reagent, as indicated. Lanes 1 and 2, T. <t>denticola</t> CF499; lanes 3 and 4, CF417; lanes 1 and 4, cell extracts; lanes 2 and 3, anti-PrtP immunoprecipitates from cell extracts. (B) T. denticola CF499 lysates were immunoprecipitated with polyclonal antibodies to PrtP, PrcA, FlaA, or Msp. Eluted proteins on duplicate blots were probed with anti-PrtP antibodies and HisProbe reagent, as indicated. (C) Immunoblot probed with anti-PrtP antibodies. Lanes 1 to 3, T. denticola CF499; lanes 4 to 6, T. denticola CF417; lanes 1 and 4, whole-cell extracts; lanes 2 and 5, anti-6×His monoclonal antibody immunoprecipitates from cell extracts; lanes 3 and 6, anti-PrtP antibody immunoprecipitates from cell extracts. (D) E. coli cells expressing PrtP, 6×His-PrcB, or both PrtP and 6×His-PrcB were analyzed directly and after Ni 2+ chromatography. Immunoblots of whole-cell extracts and Ni 2+ affinity column eluates were probed with anti-6×His monoclonal antibody or anti-PrtP polyclonal antibodies. Lanes: 1, E. coli /pCF411 (expresses PrtP); 2, E. coli /pCF415 (expresses PrcB-6×His); 3, E. coli /pCF416 (expresses PrcB-6×His + PrtP).
    T Denticola Atcc 33405, supplied by ATCC, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Construction of T. denticola mutants using ermF-ermB and ermB cassettes. The protease operon ( prcB-prcA-prtP ) and adjacent gene TDE0759 in wildtype parent strain ATCC 35405 and isogenic strains mutated in prcB (P0760 and CF548) and prcA (PNE and CF547) showing locations of ermF-ermB and ermB insertions. The symbol x- indicates gene disruption in prcA or prcB .

    Journal: Journal of microbiological methods

    Article Title: A simplified erythromycin resistance cassette for Treponema denticola mutagenesis

    doi: 10.1016/j.mimet.2010.07.020

    Figure Lengend Snippet: Construction of T. denticola mutants using ermF-ermB and ermB cassettes. The protease operon ( prcB-prcA-prtP ) and adjacent gene TDE0759 in wildtype parent strain ATCC 35405 and isogenic strains mutated in prcB (P0760 and CF548) and prcA (PNE and CF547) showing locations of ermF-ermB and ermB insertions. The symbol x- indicates gene disruption in prcA or prcB .

    Article Snippet: T. denticola ATCC 35405 (the Type strain, which has been passaged extensively in various laboratories) was electroporated with the resulting linear DNA s and plated in NOS-GN soft agar medium containing erythromycin (40 μg ml−1 , Sigma Chemical Co., St. Louis, MO) as described previously.

    Techniques:

    Southern blot analysis of tepA1 ( a ), tepA2 ( b ) , and tepA3 ( c ). Genomic DNA from T. denticola strains was digested with Hin dIII. 1: genomic DNA from ATCC 33520, 2: genomic DNA from ATCC 33521, genomic DNA from ATCC 35404, 4: genomic DNA from ATCC 35405, 5: genomic DNA from GM1

    Journal: BMC Oral Health

    Article Title: Characterization of a potential ABC-type bacteriocin exporter protein from Treponema denticola

    doi: 10.1186/s12903-016-0243-7

    Figure Lengend Snippet: Southern blot analysis of tepA1 ( a ), tepA2 ( b ) , and tepA3 ( c ). Genomic DNA from T. denticola strains was digested with Hin dIII. 1: genomic DNA from ATCC 33520, 2: genomic DNA from ATCC 33521, genomic DNA from ATCC 35404, 4: genomic DNA from ATCC 35405, 5: genomic DNA from GM1

    Article Snippet: Msps are 53–63 kDa and the identity of the amino acid sequences between T. denticola strains ATCC 35405 and OKT is 43 % [ ].

    Techniques: Southern Blot

    Binding of soluble host proteins to T. denticola cells and proteins. (A) ELISA format. FN, plasminogen (Pgn), or BSA in PBS were added to wells containing immobilized T. denticola ATCC 35405 cells (T.d.), native OppA (OppA), recombinant LacZ-OppA fusion protein (rOppA40), recombinant Msp (rMsp), or BSA. Bound FN and plasminogen were detected with anti-FN (shaded bars) and anti-plasminogen (hatched bars) antibodies, respectively. The y axis shows absorbance at 405 nm. Data shown represent the means ± standard deviations of at least two experiments using triplicate samples. (B) Ligand blot format. Duplicate samples of protein (1 μg) or cells (0.1 ml, OD 600 = 0.2) applied to nitrocellulose membranes were probed with plasminogen (samples 2 to 4) or plasminogen plus EACA (sample 5). Bound plasminogen was detected with anti-plasminogen immunoglobulin G. Lanes: 1, Plasminogen; 2, 5, and 6, T. denticola ATCC 35405; 3, T. denticola OHE; 4, T. denticola MHE.

    Journal: Infection and Immunity

    Article Title: Identification of a Treponema denticola OppA Homologue That Binds Host Proteins Present in the Subgingival Environment

    doi:

    Figure Lengend Snippet: Binding of soluble host proteins to T. denticola cells and proteins. (A) ELISA format. FN, plasminogen (Pgn), or BSA in PBS were added to wells containing immobilized T. denticola ATCC 35405 cells (T.d.), native OppA (OppA), recombinant LacZ-OppA fusion protein (rOppA40), recombinant Msp (rMsp), or BSA. Bound FN and plasminogen were detected with anti-FN (shaded bars) and anti-plasminogen (hatched bars) antibodies, respectively. The y axis shows absorbance at 405 nm. Data shown represent the means ± standard deviations of at least two experiments using triplicate samples. (B) Ligand blot format. Duplicate samples of protein (1 μg) or cells (0.1 ml, OD 600 = 0.2) applied to nitrocellulose membranes were probed with plasminogen (samples 2 to 4) or plasminogen plus EACA (sample 5). Bound plasminogen was detected with anti-plasminogen immunoglobulin G. Lanes: 1, Plasminogen; 2, 5, and 6, T. denticola ATCC 35405; 3, T. denticola OHE; 4, T. denticola MHE.

    Article Snippet: As shown in Fig. , the 70-kDa protein was released from T. denticola ATCC 35405 cells by mild treatment with Triton X-114 and, upon sequential phase partitioning at 37°C, localized to the hydrophobic detergent phase of the extracted material.

    Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant

    Confirmation of construction of allelic replacement mutants in the opp locus. (A) Southern blots of Sca I-digested T. denticola genomic DNA were probed with a biotinylated internal oppA fragment ( opp ) and the 2.1-kb ermF/AM cassette ( erm ). The ermF/AM cassette contains a single Sca I site. (B) Western immunoblot of T. denticola lysates probed with anti-70 antibodies. Lanes: 1, parent strain ATCC 35405; 2, oppA mutant strain OHE; 3, oppF mutant strain OXE.

    Journal: Infection and Immunity

    Article Title: Identification of a Treponema denticola OppA Homologue That Binds Host Proteins Present in the Subgingival Environment

    doi:

    Figure Lengend Snippet: Confirmation of construction of allelic replacement mutants in the opp locus. (A) Southern blots of Sca I-digested T. denticola genomic DNA were probed with a biotinylated internal oppA fragment ( opp ) and the 2.1-kb ermF/AM cassette ( erm ). The ermF/AM cassette contains a single Sca I site. (B) Western immunoblot of T. denticola lysates probed with anti-70 antibodies. Lanes: 1, parent strain ATCC 35405; 2, oppA mutant strain OHE; 3, oppF mutant strain OXE.

    Article Snippet: As shown in Fig. , the 70-kDa protein was released from T. denticola ATCC 35405 cells by mild treatment with Triton X-114 and, upon sequential phase partitioning at 37°C, localized to the hydrophobic detergent phase of the extracted material.

    Techniques: Western Blot, Mutagenesis

    Conservation of oppA in oral Treponema strains. (A) Southern blot of Hin dIII-digested chromosomal DNA probed with a 0.9-kb internal fragment of oppA . (B) Western blot of whole-cell extracts probed with anti-40 antibodies. Lanes: 1, T. denticola ATCC 35405; 2, T. denticola ATCC 35404; 3, T. denticola ATCC 33520; 4, T. vincentii LA-1; 5, T. denticola OTK; 6, T. denticola GM-1; 7, T. socranskii ATCC 35536; 8, T. pectinovorum ATCC 33768.

    Journal: Infection and Immunity

    Article Title: Identification of a Treponema denticola OppA Homologue That Binds Host Proteins Present in the Subgingival Environment

    doi:

    Figure Lengend Snippet: Conservation of oppA in oral Treponema strains. (A) Southern blot of Hin dIII-digested chromosomal DNA probed with a 0.9-kb internal fragment of oppA . (B) Western blot of whole-cell extracts probed with anti-40 antibodies. Lanes: 1, T. denticola ATCC 35405; 2, T. denticola ATCC 35404; 3, T. denticola ATCC 33520; 4, T. vincentii LA-1; 5, T. denticola OTK; 6, T. denticola GM-1; 7, T. socranskii ATCC 35536; 8, T. pectinovorum ATCC 33768.

    Article Snippet: As shown in Fig. , the 70-kDa protein was released from T. denticola ATCC 35405 cells by mild treatment with Triton X-114 and, upon sequential phase partitioning at 37°C, localized to the hydrophobic detergent phase of the extracted material.

    Techniques: Southern Blot, Western Blot

    Motility of wild-type T. denticola ATCC 35405 in KBM for epithelial cell lines in a 5% CO 2 atmosphere at 35°C (○) and under anaerobic conditions (85% N 2 , 10% H 2 , 5% CO 2 ) at 35°C (□). Values for each condition were obtained in triplicate in two independent experiments. More than 600 cells were examined for cellular motility in each experiment. All mutant derivatives except the nonmotile strain HL53 showed similar results (data not shown).

    Journal: Infection and Immunity

    Article Title: Motility and Chemotaxis in Tissue Penetration of Oral Epithelial Cell Layers by Treponema denticola

    doi: 10.1128/IAI.69.10.6276-6283.2001

    Figure Lengend Snippet: Motility of wild-type T. denticola ATCC 35405 in KBM for epithelial cell lines in a 5% CO 2 atmosphere at 35°C (○) and under anaerobic conditions (85% N 2 , 10% H 2 , 5% CO 2 ) at 35°C (□). Values for each condition were obtained in triplicate in two independent experiments. More than 600 cells were examined for cellular motility in each experiment. All mutant derivatives except the nonmotile strain HL53 showed similar results (data not shown).

    Article Snippet: Wild-type T. denticola ATCC 35405 was found to penetrate the tissue layers effectively, whereas a nonmotile mutant was unable to overcome the tissue barrier.

    Techniques: Mutagenesis

    Correlation of penetration rate and tissue resistance. Penetration rates were determined after 8 h of coincubation of the tissues with wild-type T. denticola . Tissue resistance was measured before and after the experiment.

    Journal: Infection and Immunity

    Article Title: Motility and Chemotaxis in Tissue Penetration of Oral Epithelial Cell Layers by Treponema denticola

    doi: 10.1128/IAI.69.10.6276-6283.2001

    Figure Lengend Snippet: Correlation of penetration rate and tissue resistance. Penetration rates were determined after 8 h of coincubation of the tissues with wild-type T. denticola . Tissue resistance was measured before and after the experiment.

    Article Snippet: Wild-type T. denticola ATCC 35405 was found to penetrate the tissue layers effectively, whereas a nonmotile mutant was unable to overcome the tissue barrier.

    Techniques:

    Tissue penetration rates (A) and motility (B) of wild-type T. denticola (Td), T. pallidum (Tp), and T. phagedenis (Tph) and various T. denticola motility and chemotaxis mutant strains. Values were determined after 8 h of coincubation of the bacteria with the tissue. Tissues used for these experiments exhibited resistances between 11 and 14Ω. More than 600 cells of each strain were examined for cellular motility.

    Journal: Infection and Immunity

    Article Title: Motility and Chemotaxis in Tissue Penetration of Oral Epithelial Cell Layers by Treponema denticola

    doi: 10.1128/IAI.69.10.6276-6283.2001

    Figure Lengend Snippet: Tissue penetration rates (A) and motility (B) of wild-type T. denticola (Td), T. pallidum (Tp), and T. phagedenis (Tph) and various T. denticola motility and chemotaxis mutant strains. Values were determined after 8 h of coincubation of the bacteria with the tissue. Tissues used for these experiments exhibited resistances between 11 and 14Ω. More than 600 cells of each strain were examined for cellular motility.

    Article Snippet: Wild-type T. denticola ATCC 35405 was found to penetrate the tissue layers effectively, whereas a nonmotile mutant was unable to overcome the tissue barrier.

    Techniques: Chemotaxis Assay, Mutagenesis

    Detection of proteins on the surface of Treponema denticola ATCC 35405 using antibodies to the Treponema pallidum protein Tp0155 which had first been pre-adsorbed against Treponema phagedenis (Kazan). (A, C, E, G, I, K and M) Phase-contrast images; (B, D, F, H, J, L and N) corresponding immunofluorescence. (A, B) Preimmune serum for Tp0155-specific antibodies; (C, D), Tp0155-specific antibodies, non-detergent-treated T. denticola ; (E, F), Tp0155-specific antibodies, Triton X-114-treated T. denticola ; (G, H), preimmune serum for Tp0249-specific antibodies; (I, J), Tp0249-specific antibodies, non-detergent-treated T. denticola (negative control); (K, L), Tp0249-specific antibodies, Triton X-114-treated T. denticola ; (M, N), anti- T. denticola ).

    Journal: Molecular oral microbiology

    Article Title: Characterization of a novel family of fibronectin-binding proteins with M23 peptidase domains from Treponema denticola

    doi: 10.1111/j.2041-1014.2010.00584.x

    Figure Lengend Snippet: Detection of proteins on the surface of Treponema denticola ATCC 35405 using antibodies to the Treponema pallidum protein Tp0155 which had first been pre-adsorbed against Treponema phagedenis (Kazan). (A, C, E, G, I, K and M) Phase-contrast images; (B, D, F, H, J, L and N) corresponding immunofluorescence. (A, B) Preimmune serum for Tp0155-specific antibodies; (C, D), Tp0155-specific antibodies, non-detergent-treated T. denticola ; (E, F), Tp0155-specific antibodies, Triton X-114-treated T. denticola ; (G, H), preimmune serum for Tp0249-specific antibodies; (I, J), Tp0249-specific antibodies, non-detergent-treated T. denticola (negative control); (K, L), Tp0249-specific antibodies, Triton X-114-treated T. denticola ; (M, N), anti- T. denticola ).

    Article Snippet: Comparisons between the T. pallidum Nichols and T. denticola ATCC 35405 genomes have become possible because they have both been fully sequenced and annotated.

    Techniques: Immunofluorescence, Negative Control

    Map of the T. denticola opp locus. The 8-kb fragment of T. denticola DNA carried on pMT1 is shown as an open bar, with the locations of some identified restriction enzyme sites shown. Immediately below are arrows representing locations of the genes identified in the opp locus. The solid arrow extending beyond the 8-kb insert represents oppA , while the shaded arrows downstream represent oppB , - C , - D , and - F . Below oppA , the solid arrow represents DNA encoding the 40-kDa LacZ-OppA fusion.

    Journal: Infection and Immunity

    Article Title: Identification of a Treponema denticola OppA Homologue That Binds Host Proteins Present in the Subgingival Environment

    doi:

    Figure Lengend Snippet: Map of the T. denticola opp locus. The 8-kb fragment of T. denticola DNA carried on pMT1 is shown as an open bar, with the locations of some identified restriction enzyme sites shown. Immediately below are arrows representing locations of the genes identified in the opp locus. The solid arrow extending beyond the 8-kb insert represents oppA , while the shaded arrows downstream represent oppB , - C , - D , and - F . Below oppA , the solid arrow represents DNA encoding the 40-kDa LacZ-OppA fusion.

    Article Snippet: A plasmid library of T. denticola ATCC 35405 genomic DNA was probed for the expression of peptides recognized by antibodies raised against the purified 70-kDa protein (anti-70), and a clone expressing high levels of an immunoreactive 40-kDa peptide was identified.

    Techniques:

    Silver-stained polyacrylamide gel of purified native ATCC 35405 Msp. Lane 1, unheated sample; lane 2, boiled sample. Arrows indicate oligomeric Msp (lane 1, 150 to 200 kDa) and monomeric Msp (lane 2, approximately 53 kDa). This preparation was used to raise rabbit antibodies against native Msp.

    Journal: Journal of Bacteriology

    Article Title: Immunotopological Analysis of the Treponema denticola Major Surface Protein (Msp)

    doi: 10.1128/JB.00528-18

    Figure Lengend Snippet: Silver-stained polyacrylamide gel of purified native ATCC 35405 Msp. Lane 1, unheated sample; lane 2, boiled sample. Arrows indicate oligomeric Msp (lane 1, 150 to 200 kDa) and monomeric Msp (lane 2, approximately 53 kDa). This preparation was used to raise rabbit antibodies against native Msp.

    Article Snippet: First, we identified a surface-exposed epitope of T. denticola ATCC 35405 Msp within a 23-residue domain in the central region of the protein that lies between the predicted Msp N-terminal (pfam02707, residues 109 to 208) and C-terminal (pfam02722, residues 360 to 543) domains.

    Techniques: Staining, Purification

    Immunofluorescence microscopy of T. denticola , revealing the Msp surface domain. T. denticola ATCC 35405 and ATCC 33520, grown to an OD 600 of 0.2, were fixed on glass slides with 1% paraformaldehyde, incubated with PBS or PBS plus 0.5% Triton X-100, and probed with rabbit antibodies to native ATCC 35405 Msp or recombinant FlaA, followed by goat anti-rabbit IgG conjugated to Alexa Fluor 568 (Msp) or Alexa Fluor 488 (FlaA), or with DAPI. Images were obtained at a magnification of 600× using an Olympus BX40 microscope fitted with an Olympus DP73 camera.

    Journal: Journal of Bacteriology

    Article Title: Immunotopological Analysis of the Treponema denticola Major Surface Protein (Msp)

    doi: 10.1128/JB.00528-18

    Figure Lengend Snippet: Immunofluorescence microscopy of T. denticola , revealing the Msp surface domain. T. denticola ATCC 35405 and ATCC 33520, grown to an OD 600 of 0.2, were fixed on glass slides with 1% paraformaldehyde, incubated with PBS or PBS plus 0.5% Triton X-100, and probed with rabbit antibodies to native ATCC 35405 Msp or recombinant FlaA, followed by goat anti-rabbit IgG conjugated to Alexa Fluor 568 (Msp) or Alexa Fluor 488 (FlaA), or with DAPI. Images were obtained at a magnification of 600× using an Olympus BX40 microscope fitted with an Olympus DP73 camera.

    Article Snippet: First, we identified a surface-exposed epitope of T. denticola ATCC 35405 Msp within a 23-residue domain in the central region of the protein that lies between the predicted Msp N-terminal (pfam02707, residues 109 to 208) and C-terminal (pfam02722, residues 360 to 543) domains.

    Techniques: Immunofluorescence, Microscopy, Incubation, Recombinant

    Map and immunoblots of deletion mutations made in msp for expression in E. coli . (Upper) The 543-residue Msp protein, with the signal peptide (SP) cleavage site indicated (residue 20), is shown above the deletions made in the coding region. The amino acid residues deleted in the N-terminal region in each of the three constructs are indicated. Deletions in the central region within the approximate boundaries of the central domain (D1 through D2) include predicted antigenic domains (Ag1 and Ag2), the remaining region of greatest divergence between Msps of strains ATCC 35405 and ATCC 33520 (Xr), and D2. These mutant Msps were expressed in E. coli . (Lower) Immunoblots of E. coli strains expressing full-length Msps with the indicated Msp residues deleted are shown. T. denticola ATCC 35405 serves as a positive control. Blots were probed with rabbit polyclonal antibodies raised against native ATCC 35405 Msp, the Msp N-terminal domain, or whole T. denticola (Td) cells. All samples were boiled prior to electrophoresis. Lanes contain lysates of E. coli strains. Antibodies raised against native T. denticola ATCC 35405 Msp or whole T. denticola ATCC 35405 cells do not recognize recombinant Msps lacking residues 229 to 251.

    Journal: Journal of Bacteriology

    Article Title: Immunotopological Analysis of the Treponema denticola Major Surface Protein (Msp)

    doi: 10.1128/JB.00528-18

    Figure Lengend Snippet: Map and immunoblots of deletion mutations made in msp for expression in E. coli . (Upper) The 543-residue Msp protein, with the signal peptide (SP) cleavage site indicated (residue 20), is shown above the deletions made in the coding region. The amino acid residues deleted in the N-terminal region in each of the three constructs are indicated. Deletions in the central region within the approximate boundaries of the central domain (D1 through D2) include predicted antigenic domains (Ag1 and Ag2), the remaining region of greatest divergence between Msps of strains ATCC 35405 and ATCC 33520 (Xr), and D2. These mutant Msps were expressed in E. coli . (Lower) Immunoblots of E. coli strains expressing full-length Msps with the indicated Msp residues deleted are shown. T. denticola ATCC 35405 serves as a positive control. Blots were probed with rabbit polyclonal antibodies raised against native ATCC 35405 Msp, the Msp N-terminal domain, or whole T. denticola (Td) cells. All samples were boiled prior to electrophoresis. Lanes contain lysates of E. coli strains. Antibodies raised against native T. denticola ATCC 35405 Msp or whole T. denticola ATCC 35405 cells do not recognize recombinant Msps lacking residues 229 to 251.

    Article Snippet: First, we identified a surface-exposed epitope of T. denticola ATCC 35405 Msp within a 23-residue domain in the central region of the protein that lies between the predicted Msp N-terminal (pfam02707, residues 109 to 208) and C-terminal (pfam02722, residues 360 to 543) domains.

    Techniques: Western Blot, Expressing, Construct, Mutagenesis, Positive Control, Electrophoresis, Recombinant

    Alignment of the Msp central domains of strains ATCC 35405 and ATCC 33520. Amino acid residue numbers are shown at the sequence ends. Purple-highlighted residues represent areas of high predicted antigenic index values (Ag1 and Ag2). Green-highlighted residues represent approximate boundaries of the central domain (D1 and D2). Blue-highlighted residues represent the region of greatest difference between the Msps other than Ag1 and Ag2 (Xr). Other than the region shown here, the amino acid sequences of the two proteins are identical, with 543 and 547 residues, respectively.

    Journal: Journal of Bacteriology

    Article Title: Immunotopological Analysis of the Treponema denticola Major Surface Protein (Msp)

    doi: 10.1128/JB.00528-18

    Figure Lengend Snippet: Alignment of the Msp central domains of strains ATCC 35405 and ATCC 33520. Amino acid residue numbers are shown at the sequence ends. Purple-highlighted residues represent areas of high predicted antigenic index values (Ag1 and Ag2). Green-highlighted residues represent approximate boundaries of the central domain (D1 and D2). Blue-highlighted residues represent the region of greatest difference between the Msps other than Ag1 and Ag2 (Xr). Other than the region shown here, the amino acid sequences of the two proteins are identical, with 543 and 547 residues, respectively.

    Article Snippet: First, we identified a surface-exposed epitope of T. denticola ATCC 35405 Msp within a 23-residue domain in the central region of the protein that lies between the predicted Msp N-terminal (pfam02707, residues 109 to 208) and C-terminal (pfam02722, residues 360 to 543) domains.

    Techniques: Sequencing

    Models for T. denticola ATCC 35405 Msp generated by I-TASSER. The I-TASSER algorithm generates 5 predicted structural models based on several parameters, of which model 1 is the overall best model. Overall model ranking does not necessarily follow the C scores for proteins lacking closely related known structures. All 5 models are displayed to show the N termini (blue) and C termini (red). The ATCC 35405 Msp surface-exposed epitope (residues 229 to 251) is highlighted in green in each model. (A) Models 1 to 5 generated using the PDB data available in June 2017. (B) Models 1 to 5 generated using the PDB data available in October 2018.

    Journal: Journal of Bacteriology

    Article Title: Immunotopological Analysis of the Treponema denticola Major Surface Protein (Msp)

    doi: 10.1128/JB.00528-18

    Figure Lengend Snippet: Models for T. denticola ATCC 35405 Msp generated by I-TASSER. The I-TASSER algorithm generates 5 predicted structural models based on several parameters, of which model 1 is the overall best model. Overall model ranking does not necessarily follow the C scores for proteins lacking closely related known structures. All 5 models are displayed to show the N termini (blue) and C termini (red). The ATCC 35405 Msp surface-exposed epitope (residues 229 to 251) is highlighted in green in each model. (A) Models 1 to 5 generated using the PDB data available in June 2017. (B) Models 1 to 5 generated using the PDB data available in October 2018.

    Article Snippet: First, we identified a surface-exposed epitope of T. denticola ATCC 35405 Msp within a 23-residue domain in the central region of the protein that lies between the predicted Msp N-terminal (pfam02707, residues 109 to 208) and C-terminal (pfam02722, residues 360 to 543) domains.

    Techniques: Generated

    Histogram of fluorescence intensity of rhodamine-phalloidin-labeled F-actin in HGF during a 140-min time course, analyzed by confocal microscopy. There was significant depolymerization of F-actin in the T. denticola ATCC 35405-challenged HGF (left graph) in comparison with the control HGF (right graph). The degree of diminished fluorescence in T. denticola -challenged cells was significant at 60 min ( P

    Journal: Infection and Immunity

    Article Title: Filamentous Actin Disruption and Diminished Inositol Phosphate Response in Gingival Fibroblasts Caused by Treponema denticola

    doi:

    Figure Lengend Snippet: Histogram of fluorescence intensity of rhodamine-phalloidin-labeled F-actin in HGF during a 140-min time course, analyzed by confocal microscopy. There was significant depolymerization of F-actin in the T. denticola ATCC 35405-challenged HGF (left graph) in comparison with the control HGF (right graph). The degree of diminished fluorescence in T. denticola -challenged cells was significant at 60 min ( P

    Article Snippet: The mean fluorescence per cell was significantly less for T. denticola ATCC 35405-challenged HGF than for control HGF by 60 min and all subsequent time periods ([2.15 ± 0.24] × 106 [mean ± standard error of mean] for T. denticola and [3.32 ± 0.34] × 106 for control at 60 min; P < 0.01 [Fig. ]).

    Techniques: Fluorescence, Labeling, Confocal Microscopy

    Comparison of filamentous actin disruption (mean ± standard deviation) in HGF challenged with different concentrations of whole cells of T. denticola ATCC 35405 (□) and OM extract (▪). The dry weight for the OM and the dry weight for the T. denticola cells were equivalent at each optical density (OD) point shown on the x axis.

    Journal: Infection and Immunity

    Article Title: Filamentous Actin Disruption and Diminished Inositol Phosphate Response in Gingival Fibroblasts Caused by Treponema denticola

    doi:

    Figure Lengend Snippet: Comparison of filamentous actin disruption (mean ± standard deviation) in HGF challenged with different concentrations of whole cells of T. denticola ATCC 35405 (□) and OM extract (▪). The dry weight for the OM and the dry weight for the T. denticola cells were equivalent at each optical density (OD) point shown on the x axis.

    Article Snippet: The mean fluorescence per cell was significantly less for T. denticola ATCC 35405-challenged HGF than for control HGF by 60 min and all subsequent time periods ([2.15 ± 0.24] × 106 [mean ± standard error of mean] for T. denticola and [3.32 ± 0.34] × 106 for control at 60 min; P < 0.01 [Fig. ]).

    Techniques: Standard Deviation

    Qualitative comparison by SDS-PAGE of T. denticola ATCC 35405 whole cells (lane 1), OM extract run at the same time (lane 2), and repeat of the OM extract run in a different gel to resolve mid-range bands more clearly (lane 3). The positions of molecular size standards (broad-range SDS-PAGE standards; Bio-Rad) are shown in kilodaltons at the left. Conditions were as follows: Washed T. denticola suspensions (optical density of 1.2 at 550 nm) from 3-day cultures or undiluted OM extract were mixed 1:1 with sample buffer (0.25 M Tris-HCl [pH 6.8], 20% glycerol, 4.6% SDS, 0.02% bromophenol blue, 10% β-mercaptoethanol) and boiled for 10 min; 25 μl was added to each lane of a 1-mm-thick Ready Gel (Bio-Rad) with a 4% stacking gel and 12% resolving gel. Electrophoresis was run in a Mini-Protean II cell (Bio-Rad). Whole cells and OM extract had several bands in common.

    Journal: Infection and Immunity

    Article Title: Filamentous Actin Disruption and Diminished Inositol Phosphate Response in Gingival Fibroblasts Caused by Treponema denticola

    doi:

    Figure Lengend Snippet: Qualitative comparison by SDS-PAGE of T. denticola ATCC 35405 whole cells (lane 1), OM extract run at the same time (lane 2), and repeat of the OM extract run in a different gel to resolve mid-range bands more clearly (lane 3). The positions of molecular size standards (broad-range SDS-PAGE standards; Bio-Rad) are shown in kilodaltons at the left. Conditions were as follows: Washed T. denticola suspensions (optical density of 1.2 at 550 nm) from 3-day cultures or undiluted OM extract were mixed 1:1 with sample buffer (0.25 M Tris-HCl [pH 6.8], 20% glycerol, 4.6% SDS, 0.02% bromophenol blue, 10% β-mercaptoethanol) and boiled for 10 min; 25 μl was added to each lane of a 1-mm-thick Ready Gel (Bio-Rad) with a 4% stacking gel and 12% resolving gel. Electrophoresis was run in a Mini-Protean II cell (Bio-Rad). Whole cells and OM extract had several bands in common.

    Article Snippet: The mean fluorescence per cell was significantly less for T. denticola ATCC 35405-challenged HGF than for control HGF by 60 min and all subsequent time periods ([2.15 ± 0.24] × 106 [mean ± standard error of mean] for T. denticola and [3.32 ± 0.34] × 106 for control at 60 min; P < 0.01 [Fig. ]).

    Techniques: SDS Page, Nucleic Acid Electrophoresis

    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.

    Journal: Molecular Microbiology

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

    doi: 10.1111/j.1365-2958.2008.06418.x

    Figure Lengend 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.

    Article Snippet: Identification of the tro operon in T. denticola Analysis of the T. denticola ATCC 35405 genome sequence revealed the presence of a genetic locus demonstrating significant similarity to the T. pallidum tro operon.

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

    Mean areas of lesions at infection sites after challenge with T. denticola ATCC 35405 and prtP mutant K1. Mice were injected with live T. denticola ATCC 35405 (open bars) or mutant K1 (hatched bars), and lesion areas were determined at the indicated times following infection.

    Journal: Journal of Bacteriology

    Article Title: Dentilisin Activity Affects the Organization of the Outer Sheath of Treponema denticola

    doi:

    Figure Lengend Snippet: Mean areas of lesions at infection sites after challenge with T. denticola ATCC 35405 and prtP mutant K1. Mice were injected with live T. denticola ATCC 35405 (open bars) or mutant K1 (hatched bars), and lesion areas were determined at the indicated times following infection.

    Article Snippet: We analyzed the antigenic proteins with antiserum against T. denticola ATCC 35405 whole cells (Fig. ).

    Techniques: Infection, Mutagenesis, Mouse Assay, Injection

    Immunoblot analysis of T. denticola ATCC 35405 and K1 with anti- T. denticola ATCC 35405 whole-cell serum. Samples from lanes 1 to 4 were treated with PMSF. Lanes 1 and 5, T. denticola ATCC 35405 (without boiling); lanes 2 and 6, T. denticola K1 (without boiling); lanes 3 and 7, T. denticola ATCC 35405 (with boiling); lanes 4 and 8, T. denticola K1 (with boiling). Arrowheads indicate the high-molecular-mass oligomeric protein and the Msp band.

    Journal: Journal of Bacteriology

    Article Title: Dentilisin Activity Affects the Organization of the Outer Sheath of Treponema denticola

    doi:

    Figure Lengend Snippet: Immunoblot analysis of T. denticola ATCC 35405 and K1 with anti- T. denticola ATCC 35405 whole-cell serum. Samples from lanes 1 to 4 were treated with PMSF. Lanes 1 and 5, T. denticola ATCC 35405 (without boiling); lanes 2 and 6, T. denticola K1 (without boiling); lanes 3 and 7, T. denticola ATCC 35405 (with boiling); lanes 4 and 8, T. denticola K1 (with boiling). Arrowheads indicate the high-molecular-mass oligomeric protein and the Msp band.

    Article Snippet: We analyzed the antigenic proteins with antiserum against T. denticola ATCC 35405 whole cells (Fig. ).

    Techniques:

    Southern blot analysis of T. denticola ATCC 35405 and prtP mutant K1. Chromosomal DNAs from T. denticola ATCC 35405 (lanes 1 and 3) and prtP mutant K1 (lanes 2 and 4) were digested with Hin dIII and hybridized with a digoxigenin-labeled Kpn I- Pst I fragment from the prtP gene (lanes 1 and 2) or the Kpn I- Bam HI fragment from pVA2198 (lanes 3 and 4). Numbers at right are kilobase pairs.

    Journal: Journal of Bacteriology

    Article Title: Dentilisin Activity Affects the Organization of the Outer Sheath of Treponema denticola

    doi:

    Figure Lengend Snippet: Southern blot analysis of T. denticola ATCC 35405 and prtP mutant K1. Chromosomal DNAs from T. denticola ATCC 35405 (lanes 1 and 3) and prtP mutant K1 (lanes 2 and 4) were digested with Hin dIII and hybridized with a digoxigenin-labeled Kpn I- Pst I fragment from the prtP gene (lanes 1 and 2) or the Kpn I- Bam HI fragment from pVA2198 (lanes 3 and 4). Numbers at right are kilobase pairs.

    Article Snippet: We analyzed the antigenic proteins with antiserum against T. denticola ATCC 35405 whole cells (Fig. ).

    Techniques: Southern Blot, Mutagenesis, Labeling

    Gelatin zymography of sonicates of T. denticola ATCC 35405 (lane 1) and K1 (lane 2).

    Journal: Journal of Bacteriology

    Article Title: Dentilisin Activity Affects the Organization of the Outer Sheath of Treponema denticola

    doi:

    Figure Lengend Snippet: Gelatin zymography of sonicates of T. denticola ATCC 35405 (lane 1) and K1 (lane 2).

    Article Snippet: We analyzed the antigenic proteins with antiserum against T. denticola ATCC 35405 whole cells (Fig. ).

    Techniques: Zymography

    SDS-PAGE analysis of sonicates of T. denticola ATCC 35405 and K1. Samples from lanes 1 to 4 were treated with PMSF. Lanes 1 and 5, T. denticola ATCC 35405 (without boiling); lanes 2 and 6, T. denticola K1 (without boiling); lanes 3 and 7, T. denticola ATCC 35405 (with boiling); lanes 4 and 8, T. denticola K1 (with boiling). After electrophoresis, the gel was stained with Coomassie brilliant blue R-250. Arrowheads indicate the high-molecular-mass oligomeric protein and the Msp band.

    Journal: Journal of Bacteriology

    Article Title: Dentilisin Activity Affects the Organization of the Outer Sheath of Treponema denticola

    doi:

    Figure Lengend Snippet: SDS-PAGE analysis of sonicates of T. denticola ATCC 35405 and K1. Samples from lanes 1 to 4 were treated with PMSF. Lanes 1 and 5, T. denticola ATCC 35405 (without boiling); lanes 2 and 6, T. denticola K1 (without boiling); lanes 3 and 7, T. denticola ATCC 35405 (with boiling); lanes 4 and 8, T. denticola K1 (with boiling). After electrophoresis, the gel was stained with Coomassie brilliant blue R-250. Arrowheads indicate the high-molecular-mass oligomeric protein and the Msp band.

    Article Snippet: We analyzed the antigenic proteins with antiserum against T. denticola ATCC 35405 whole cells (Fig. ).

    Techniques: SDS Page, Electrophoresis, Staining

    Immunoblot analysis of T. denticola ATCC 35405 (lane 1; boiled) and K1 (lane 2; boiled) with antidentilisin serum.

    Journal: Journal of Bacteriology

    Article Title: Dentilisin Activity Affects the Organization of the Outer Sheath of Treponema denticola

    doi:

    Figure Lengend Snippet: Immunoblot analysis of T. denticola ATCC 35405 (lane 1; boiled) and K1 (lane 2; boiled) with antidentilisin serum.

    Article Snippet: We analyzed the antigenic proteins with antiserum against T. denticola ATCC 35405 whole cells (Fig. ).

    Techniques:

    Role of the CTLP complex in the formation of T. denticola / P. gingivalis dual-species biofilms. Representative CLSM images of dual-species biofilms of (a, b) T. denticola ATCC 35405 and (c, d) T. denticola CKE, with P. gingivalis ATCC 33277. Biofilms were

    Journal: Microbiology

    Article Title: Treponema denticola chymotrypsin-like proteinase (CTLP) integrates spirochaetes within oral microbial communities

    doi: 10.1099/mic.0.055939-0

    Figure Lengend Snippet: Role of the CTLP complex in the formation of T. denticola / P. gingivalis dual-species biofilms. Representative CLSM images of dual-species biofilms of (a, b) T. denticola ATCC 35405 and (c, d) T. denticola CKE, with P. gingivalis ATCC 33277. Biofilms were

    Article Snippet: Treponema strains used in this study were T. denticola ATCC 35405 wild-type, isogenic mutant strains CKE Δ prcAprtP (CTLP-negative) ( ) and MHE Δ msp (Msp-negative) , and Treponema vincentii ATCC 35580.

    Techniques: Confocal Laser Scanning Microscopy

    Adherence levels of Treponema to other periodontopathogens. The input cell number was 1.2×10 7 cells per well of T. denticola ATCC 35405 (black bars), T. denticola MHE (mid-grey bars), T. denticola CKE (dark-grey bars) and T. vincentii ATCC 35580

    Journal: Microbiology

    Article Title: Treponema denticola chymotrypsin-like proteinase (CTLP) integrates spirochaetes within oral microbial communities

    doi: 10.1099/mic.0.055939-0

    Figure Lengend Snippet: Adherence levels of Treponema to other periodontopathogens. The input cell number was 1.2×10 7 cells per well of T. denticola ATCC 35405 (black bars), T. denticola MHE (mid-grey bars), T. denticola CKE (dark-grey bars) and T. vincentii ATCC 35580

    Article Snippet: Treponema strains used in this study were T. denticola ATCC 35405 wild-type, isogenic mutant strains CKE Δ prcAprtP (CTLP-negative) ( ) and MHE Δ msp (Msp-negative) , and Treponema vincentii ATCC 35580.

    Techniques:

    Fluorescence microscopy of monospecies or dual-species biofilms of T. denticola ATCC 35405, T. denticola CKE or T. denticola MHE, and P. gingivalis ATCC 33277, and corresponding biomass data. (a) Biofilms formed upon saliva-coated coverslips after 72

    Journal: Microbiology

    Article Title: Treponema denticola chymotrypsin-like proteinase (CTLP) integrates spirochaetes within oral microbial communities

    doi: 10.1099/mic.0.055939-0

    Figure Lengend Snippet: Fluorescence microscopy of monospecies or dual-species biofilms of T. denticola ATCC 35405, T. denticola CKE or T. denticola MHE, and P. gingivalis ATCC 33277, and corresponding biomass data. (a) Biofilms formed upon saliva-coated coverslips after 72

    Article Snippet: Treponema strains used in this study were T. denticola ATCC 35405 wild-type, isogenic mutant strains CKE Δ prcAprtP (CTLP-negative) ( ) and MHE Δ msp (Msp-negative) , and Treponema vincentii ATCC 35580.

    Techniques: Fluorescence, Microscopy

    T. denticola proteases are not responsible for resistance to hβD-2. (A) T. denticola ATCC 35404 incubated in the presence or absence of 100 μM chymostatin (CHY) remained resistant to 10 μg of hβD-2 per ml over a 4-h period. (B) E. coli was incubated in the presence of 10 μg of hβD-2 per ml for 4 h, with or without equal numbers of T. denticola ATCC 35404. Data are means ± SEM from five (A) and three (B) experiments. SDS, sodium dodecyl sulfate.

    Journal: Infection and Immunity

    Article Title: Treponema denticola Is Resistant to Human ?-Defensins

    doi: 10.1128/IAI.70.7.3982-3984.2002

    Figure Lengend Snippet: T. denticola proteases are not responsible for resistance to hβD-2. (A) T. denticola ATCC 35404 incubated in the presence or absence of 100 μM chymostatin (CHY) remained resistant to 10 μg of hβD-2 per ml over a 4-h period. (B) E. coli was incubated in the presence of 10 μg of hβD-2 per ml for 4 h, with or without equal numbers of T. denticola ATCC 35404. Data are means ± SEM from five (A) and three (B) experiments. SDS, sodium dodecyl sulfate.

    Article Snippet: T. denticola ATCC 35404 and 10 μg of hβD-2 per ml were incubated in the presence or absence of final concentrations of 100 μM chymostatin (Sigma Chemicals, St. Louis, Mo.) at 37°C and 5% CO2 for 4 h. Viable bacteria were enumerated by dark-field microscopy ( T. denticola ) or plate counts ( E. coli ).

    Techniques: Incubation

    CGase amino acid sequences from T. denticola strains ATCC 35404 and 35405. The top line in each row shows the predicted amino acid sequence for CGase from strain ATCC 35404 (this paper). The second line ). The asterisks represent amino acids that are identical in the two strains. Triangles ).

    Journal: The Journal of Biological Chemistry

    Article Title: A 52-kDa Leucyl Aminopeptidase from Treponema denticola Is a Cysteinylglycinase That Mediates the Second Step of Glutathione Metabolism *

    doi: 10.1074/jbc.M801034200

    Figure Lengend Snippet: CGase amino acid sequences from T. denticola strains ATCC 35404 and 35405. The top line in each row shows the predicted amino acid sequence for CGase from strain ATCC 35404 (this paper). The second line ). The asterisks represent amino acids that are identical in the two strains. Triangles ).

    Article Snippet: To rectify this, we first showed that sonicated extracts of T. denticola strain ATCC 35404 had CGase activity ( ).

    Techniques: Sequencing

    Fractionation of 52-kDa CGase from T. denticola ATCC 35404 on a HiTrap Q FF column. a , fractions from the separation of a partially purified protein extract on a HiTrap Sepharose Q FF ion exchange column were assayed for CGase activity ( squares ) and total protein ( diamonds , as OD 280 ). b ) of the proteins present in the sample that was loaded onto the ion exchange column ( lane 1 ) and the proteins present in fraction 15 that had the peak CGase activity ( lane 2 ). Proteins of known molecular weight were run in lane MW , and the gel was stained with 0.025% Coomassie Brilliant Blue R-250.

    Journal: The Journal of Biological Chemistry

    Article Title: A 52-kDa Leucyl Aminopeptidase from Treponema denticola Is a Cysteinylglycinase That Mediates the Second Step of Glutathione Metabolism *

    doi: 10.1074/jbc.M801034200

    Figure Lengend Snippet: Fractionation of 52-kDa CGase from T. denticola ATCC 35404 on a HiTrap Q FF column. a , fractions from the separation of a partially purified protein extract on a HiTrap Sepharose Q FF ion exchange column were assayed for CGase activity ( squares ) and total protein ( diamonds , as OD 280 ). b ) of the proteins present in the sample that was loaded onto the ion exchange column ( lane 1 ) and the proteins present in fraction 15 that had the peak CGase activity ( lane 2 ). Proteins of known molecular weight were run in lane MW , and the gel was stained with 0.025% Coomassie Brilliant Blue R-250.

    Article Snippet: To rectify this, we first showed that sonicated extracts of T. denticola strain ATCC 35404 had CGase activity ( ).

    Techniques: Fractionation, Purification, Activity Assay, Molecular Weight, Staining

    Effect of dissipation of proton motive force on total association of β-defensins with T. denticola . A total of 1 × 10 8 cells/ml of T. denticola 35404 were incubated with biotinylated β-defensin 2 or 3 (10 μg/ml) and 50

    Journal:

    Article Title: Mechanisms of Decreased Susceptibility to ?-Defensins by Treponema denticola ▿

    doi: 10.1128/IAI.01718-06

    Figure Lengend Snippet: Effect of dissipation of proton motive force on total association of β-defensins with T. denticola . A total of 1 × 10 8 cells/ml of T. denticola 35404 were incubated with biotinylated β-defensin 2 or 3 (10 μg/ml) and 50

    Article Snippet: To examine this possibility, biotinylated hβD-2 or -3 was incubated with T. denticola 35404, 33520, 33521, the ATCC 35405 parent, the ATCC 35405 K1 dentilisin mutant, E. coli ML35, or S. aureus 113 dlt .

    Techniques: Incubation

    Dissipation of proton motive force increases T. denticola sensitivity to hβD-3. (A) A total of 1 × 10 8 cells/ml of T. denticola 35404 were incubated in triplicate at 37°C with 35 μM CCCP or DMSO (solvent). Then 10 μl/well

    Journal:

    Article Title: Mechanisms of Decreased Susceptibility to ?-Defensins by Treponema denticola ▿

    doi: 10.1128/IAI.01718-06

    Figure Lengend Snippet: Dissipation of proton motive force increases T. denticola sensitivity to hβD-3. (A) A total of 1 × 10 8 cells/ml of T. denticola 35404 were incubated in triplicate at 37°C with 35 μM CCCP or DMSO (solvent). Then 10 μl/well

    Article Snippet: To examine this possibility, biotinylated hβD-2 or -3 was incubated with T. denticola 35404, 33520, 33521, the ATCC 35405 parent, the ATCC 35405 K1 dentilisin mutant, E. coli ML35, or S. aureus 113 dlt .

    Techniques: Incubation

    Effect of host proteins present in the growth medium on T. denticola susceptibility to β-defensins. A total of 1 × 10 5 T. denticola 35404 cells grown in GM-1 serum-containing medium or in chemically defined medium (CDM) lacking serum,

    Journal:

    Article Title: Mechanisms of Decreased Susceptibility to ?-Defensins by Treponema denticola ▿

    doi: 10.1128/IAI.01718-06

    Figure Lengend Snippet: Effect of host proteins present in the growth medium on T. denticola susceptibility to β-defensins. A total of 1 × 10 5 T. denticola 35404 cells grown in GM-1 serum-containing medium or in chemically defined medium (CDM) lacking serum,

    Article Snippet: To examine this possibility, biotinylated hβD-2 or -3 was incubated with T. denticola 35404, 33520, 33521, the ATCC 35405 parent, the ATCC 35405 K1 dentilisin mutant, E. coli ML35, or S. aureus 113 dlt .

    Techniques:

    Quenching of a fluorescent dye in the presence of efflux pump inhibitors. A total of 1 × 10 8 cells/ml of T. denticola 35404 were incubated in 12 replicates at 37°C with 10 μg/ml reserpine, 20 μg/ml verapamil, 50 μM

    Journal:

    Article Title: Mechanisms of Decreased Susceptibility to ?-Defensins by Treponema denticola ▿

    doi: 10.1128/IAI.01718-06

    Figure Lengend Snippet: Quenching of a fluorescent dye in the presence of efflux pump inhibitors. A total of 1 × 10 8 cells/ml of T. denticola 35404 were incubated in 12 replicates at 37°C with 10 μg/ml reserpine, 20 μg/ml verapamil, 50 μM

    Article Snippet: To examine this possibility, biotinylated hβD-2 or -3 was incubated with T. denticola 35404, 33520, 33521, the ATCC 35405 parent, the ATCC 35405 K1 dentilisin mutant, E. coli ML35, or S. aureus 113 dlt .

    Techniques: Incubation

    Coaggregation and biofilm formation of T. denticola wild-type, Δ flgE , Δ motB with P. gingivalis W50. (A) Coaggregation of T. denticola wild-type and mutants with P. gingivalis W50. T. denticola ATCC 33520 and P. gingivalis cells at exponential growth phase were harvested, washed twice with coaggregation buffer and adjusted to A 650 of 0.5 with the coaggregation buffer. Equal volumes of T. denticola and P. gingivalis cell suspensions were combined. The height of bacterial aggregates was monitored for 7 h. The data are presented as means plus standard deviations ( N = 3). (B) Monospecies and dual-species static biofilms of T. denticola ATCC 33520 wild-type, Δ flgE , Δ motB with P. gingivalis W50. T. denticola and P. gingivalis cells were grown to exponential phase, diluted to A 650 of 0.15 and grown anaerobically for 5 days in a 12-well plate, either in a monospecies or dual-species culture. The resultant biofilm was stained with crystal violet and the total biomass determined spectrophotometrically. The data are presented as means and standard deviations ( N = 23–27) and were analyzed using a Kruskal-Wallis with Conover-Imam test. All values were significantly different ( p

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: The Role of Treponema denticola Motility in Synergistic Biofilm Formation With Porphyromonas gingivalis

    doi: 10.3389/fcimb.2019.00432

    Figure Lengend Snippet: Coaggregation and biofilm formation of T. denticola wild-type, Δ flgE , Δ motB with P. gingivalis W50. (A) Coaggregation of T. denticola wild-type and mutants with P. gingivalis W50. T. denticola ATCC 33520 and P. gingivalis cells at exponential growth phase were harvested, washed twice with coaggregation buffer and adjusted to A 650 of 0.5 with the coaggregation buffer. Equal volumes of T. denticola and P. gingivalis cell suspensions were combined. The height of bacterial aggregates was monitored for 7 h. The data are presented as means plus standard deviations ( N = 3). (B) Monospecies and dual-species static biofilms of T. denticola ATCC 33520 wild-type, Δ flgE , Δ motB with P. gingivalis W50. T. denticola and P. gingivalis cells were grown to exponential phase, diluted to A 650 of 0.15 and grown anaerobically for 5 days in a 12-well plate, either in a monospecies or dual-species culture. The resultant biofilm was stained with crystal violet and the total biomass determined spectrophotometrically. The data are presented as means and standard deviations ( N = 23–27) and were analyzed using a Kruskal-Wallis with Conover-Imam test. All values were significantly different ( p

    Article Snippet: Although T. denticola ATCC 33520 is commonly observed to have four periplasmic flagella (Izard et al., ), in this study up to five periplasmic flagella were occasionally observed ( ).

    Techniques: Staining

    Western blots with T. denticola Tap1 antiserum (A), T. pallidum FlgE antiserum (B), and T. phagedenis FlaB antiserum (C). Lanes 1, JS97; lanes 2, AS98; lanes 3, wild type. The arrow in panel A indicates the Tap1 polypeptide band. In panel B, the polypeptide ladder is a typical pattern found in treponemal hook polypeptides that may be due to cross-linking. The significance of the minor band missing in lane 3 is unknown. Numbers at right of each panel represent molecular masses in kilodaltons.

    Journal: Journal of Bacteriology

    Article Title: Insertional Inactivation of Treponemadenticola tap1 Results in a Nonmotile Mutant with Elongated Flagellar Hooks

    doi:

    Figure Lengend Snippet: Western blots with T. denticola Tap1 antiserum (A), T. pallidum FlgE antiserum (B), and T. phagedenis FlaB antiserum (C). Lanes 1, JS97; lanes 2, AS98; lanes 3, wild type. The arrow in panel A indicates the Tap1 polypeptide band. In panel B, the polypeptide ladder is a typical pattern found in treponemal hook polypeptides that may be due to cross-linking. The significance of the minor band missing in lane 3 is unknown. Numbers at right of each panel represent molecular masses in kilodaltons.

    Article Snippet: T. denticola (ATCC 33520) was grown in new oral spirochete medium (NOS) with 10% heat-inactivated rabbit serum and 10 μg of cocarboxylase per ml at 36°C in an anaerobic chamber (Coy Laboratory Products, Inc., Grass Lake, Mich.) with an atmosphere of 85% nitrogen, 5% carbon dioxide, and 10% hydrogen.

    Techniques: Western Blot

    Diagram of the fla operon organization of T. denticola and 5′ upstream region. P fla indicates the approximate location of the fla operon promoter. The ermF-ermAM cassette is indicated above the operon and is shown where it is inserted in the Bgl II site for creating Tap1-deficient mutants. The locations of primer pairs used for RT-PCR are indicated by arrows and are represented as follows: 1, TDW8; 2, TDW9; 3, TDW12; 4, TDW5; 5, TDWFLGEF; 6, TDWFLGER. Sequences of the primers are given in Materials and Methods.

    Journal: Journal of Bacteriology

    Article Title: Insertional Inactivation of Treponemadenticola tap1 Results in a Nonmotile Mutant with Elongated Flagellar Hooks

    doi:

    Figure Lengend Snippet: Diagram of the fla operon organization of T. denticola and 5′ upstream region. P fla indicates the approximate location of the fla operon promoter. The ermF-ermAM cassette is indicated above the operon and is shown where it is inserted in the Bgl II site for creating Tap1-deficient mutants. The locations of primer pairs used for RT-PCR are indicated by arrows and are represented as follows: 1, TDW8; 2, TDW9; 3, TDW12; 4, TDW5; 5, TDWFLGEF; 6, TDWFLGER. Sequences of the primers are given in Materials and Methods.

    Article Snippet: T. denticola (ATCC 33520) was grown in new oral spirochete medium (NOS) with 10% heat-inactivated rabbit serum and 10 μg of cocarboxylase per ml at 36°C in an anaerobic chamber (Coy Laboratory Products, Inc., Grass Lake, Mich.) with an atmosphere of 85% nitrogen, 5% carbon dioxide, and 10% hydrogen.

    Techniques: Reverse Transcription Polymerase Chain Reaction

    Comparison of the T. denticola fla promoter sequence with promoter sequences from various spirochete motility genes and consensus sigma 28 sequences.

    Journal: Journal of Bacteriology

    Article Title: Insertional Inactivation of Treponemadenticola tap1 Results in a Nonmotile Mutant with Elongated Flagellar Hooks

    doi:

    Figure Lengend Snippet: Comparison of the T. denticola fla promoter sequence with promoter sequences from various spirochete motility genes and consensus sigma 28 sequences.

    Article Snippet: T. denticola (ATCC 33520) was grown in new oral spirochete medium (NOS) with 10% heat-inactivated rabbit serum and 10 μg of cocarboxylase per ml at 36°C in an anaerobic chamber (Coy Laboratory Products, Inc., Grass Lake, Mich.) with an atmosphere of 85% nitrogen, 5% carbon dioxide, and 10% hydrogen.

    Techniques: Sequencing

    Alignment of Tap1 amino acid sequences. (A) Identical amino acids from three treponemes are boxed and shaded with SHADYBOX, which reveals the conserved region near the carboxyl terminus. The dark inverted triangle indicates the location of the point of insertion for the erythromycin resistance cassette into the T. denticola tap1 gene to generate a Tap1-deficient mutant. (B) Alignment of the conserved C-terminal region of T. denticola Tap1 with FliK of S. typhimurium ). Identical amino acids are boxed and shaded as described above.

    Journal: Journal of Bacteriology

    Article Title: Insertional Inactivation of Treponemadenticola tap1 Results in a Nonmotile Mutant with Elongated Flagellar Hooks

    doi:

    Figure Lengend Snippet: Alignment of Tap1 amino acid sequences. (A) Identical amino acids from three treponemes are boxed and shaded with SHADYBOX, which reveals the conserved region near the carboxyl terminus. The dark inverted triangle indicates the location of the point of insertion for the erythromycin resistance cassette into the T. denticola tap1 gene to generate a Tap1-deficient mutant. (B) Alignment of the conserved C-terminal region of T. denticola Tap1 with FliK of S. typhimurium ). Identical amino acids are boxed and shaded as described above.

    Article Snippet: T. denticola (ATCC 33520) was grown in new oral spirochete medium (NOS) with 10% heat-inactivated rabbit serum and 10 μg of cocarboxylase per ml at 36°C in an anaerobic chamber (Coy Laboratory Products, Inc., Grass Lake, Mich.) with an atmosphere of 85% nitrogen, 5% carbon dioxide, and 10% hydrogen.

    Techniques: Mutagenesis

    T. denticola wild type (WT) and Tap1-deficient mutant JS97 after growth for 7 days on NOS plates containing 0.5% agarose. Approximately 0.1 μl was placed on the plate and incubated at 36°C in an anaerobic chamber.

    Journal: Journal of Bacteriology

    Article Title: Insertional Inactivation of Treponemadenticola tap1 Results in a Nonmotile Mutant with Elongated Flagellar Hooks

    doi:

    Figure Lengend Snippet: T. denticola wild type (WT) and Tap1-deficient mutant JS97 after growth for 7 days on NOS plates containing 0.5% agarose. Approximately 0.1 μl was placed on the plate and incubated at 36°C in an anaerobic chamber.

    Article Snippet: T. denticola (ATCC 33520) was grown in new oral spirochete medium (NOS) with 10% heat-inactivated rabbit serum and 10 μg of cocarboxylase per ml at 36°C in an anaerobic chamber (Coy Laboratory Products, Inc., Grass Lake, Mich.) with an atmosphere of 85% nitrogen, 5% carbon dioxide, and 10% hydrogen.

    Techniques: Mutagenesis, Incubation

    Conservation of prcB in T. denticola . The deduced amino acid sequence of the complete ORF, including the PrcB coding region, is shown for T. denticola strains ATCC 35405, ATCC 33520, and OTK, aligned using Clustal 2.0. The N-terminal valine residue predicted by JCVI (Val 38 ) and the two possible N-terminal methionine residues (Met 1 and Met 7 ) predicted by the present study are shaded gray. The predicted signal peptidase II cleavage site is indicated by a black arrowhead. Amino acid identity throughout all three sequences is indicated by asterisks. Conserved and semiconserved amino acids are shown by colons and dots, respectively.

    Journal: Journal of Bacteriology

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex ▿

    doi: 10.1128/JB.00274-10

    Figure Lengend Snippet: Conservation of prcB in T. denticola . The deduced amino acid sequence of the complete ORF, including the PrcB coding region, is shown for T. denticola strains ATCC 35405, ATCC 33520, and OTK, aligned using Clustal 2.0. The N-terminal valine residue predicted by JCVI (Val 38 ) and the two possible N-terminal methionine residues (Met 1 and Met 7 ) predicted by the present study are shaded gray. The predicted signal peptidase II cleavage site is indicated by a black arrowhead. Amino acid identity throughout all three sequences is indicated by asterisks. Conserved and semiconserved amino acids are shown by colons and dots, respectively.

    Article Snippet: DNA sequences of the prcB ORF in T. denticola strains ATCC 33520 and OTK were amplified from genomic DNA with primers CX402 and CX360.

    Techniques: Sequencing

    Interaction of PrcB with PrtP. Arrows denote PrcB, PrtP, and rabbit IgG heavy chains (H). (A) Immunoblots probed with anti-PrtP antibodies and HisProbe reagent, as indicated. Lanes 1 and 2, T. denticola CF499; lanes 3 and 4, CF417; lanes 1 and 4, cell extracts; lanes 2 and 3, anti-PrtP immunoprecipitates from cell extracts. (B) T. denticola CF499 lysates were immunoprecipitated with polyclonal antibodies to PrtP, PrcA, FlaA, or Msp. Eluted proteins on duplicate blots were probed with anti-PrtP antibodies and HisProbe reagent, as indicated. (C) Immunoblot probed with anti-PrtP antibodies. Lanes 1 to 3, T. denticola CF499; lanes 4 to 6, T. denticola CF417; lanes 1 and 4, whole-cell extracts; lanes 2 and 5, anti-6×His monoclonal antibody immunoprecipitates from cell extracts; lanes 3 and 6, anti-PrtP antibody immunoprecipitates from cell extracts. (D) E. coli cells expressing PrtP, 6×His-PrcB, or both PrtP and 6×His-PrcB were analyzed directly and after Ni 2+ chromatography. Immunoblots of whole-cell extracts and Ni 2+ affinity column eluates were probed with anti-6×His monoclonal antibody or anti-PrtP polyclonal antibodies. Lanes: 1, E. coli /pCF411 (expresses PrtP); 2, E. coli /pCF415 (expresses PrcB-6×His); 3, E. coli /pCF416 (expresses PrcB-6×His + PrtP).

    Journal: Journal of Bacteriology

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex ▿

    doi: 10.1128/JB.00274-10

    Figure Lengend Snippet: Interaction of PrcB with PrtP. Arrows denote PrcB, PrtP, and rabbit IgG heavy chains (H). (A) Immunoblots probed with anti-PrtP antibodies and HisProbe reagent, as indicated. Lanes 1 and 2, T. denticola CF499; lanes 3 and 4, CF417; lanes 1 and 4, cell extracts; lanes 2 and 3, anti-PrtP immunoprecipitates from cell extracts. (B) T. denticola CF499 lysates were immunoprecipitated with polyclonal antibodies to PrtP, PrcA, FlaA, or Msp. Eluted proteins on duplicate blots were probed with anti-PrtP antibodies and HisProbe reagent, as indicated. (C) Immunoblot probed with anti-PrtP antibodies. Lanes 1 to 3, T. denticola CF499; lanes 4 to 6, T. denticola CF417; lanes 1 and 4, whole-cell extracts; lanes 2 and 5, anti-6×His monoclonal antibody immunoprecipitates from cell extracts; lanes 3 and 6, anti-PrtP antibody immunoprecipitates from cell extracts. (D) E. coli cells expressing PrtP, 6×His-PrcB, or both PrtP and 6×His-PrcB were analyzed directly and after Ni 2+ chromatography. Immunoblots of whole-cell extracts and Ni 2+ affinity column eluates were probed with anti-6×His monoclonal antibody or anti-PrtP polyclonal antibodies. Lanes: 1, E. coli /pCF411 (expresses PrtP); 2, E. coli /pCF415 (expresses PrcB-6×His); 3, E. coli /pCF416 (expresses PrcB-6×His + PrtP).

    Article Snippet: Counting from the first AUG codon in the ORF, the deduced amino acid sequences are identical for residues 1 to 65, which includes the predicted untranslated region upstream of the GUG translation initiation codon in the annotated T. denticola genome sequence ( ).

    Techniques: Western Blot, Immunoprecipitation, Expressing, Chromatography, Affinity Column

    Protease locus in T. denticola parent and prcB mutant strains. The genes of the protease operon, prcB , prcA , and prtP , are shown as gray arrows. The location of the protease operon promoter region is indicated by “P.” The protease mRNA transcript is shown as a thin arrow above the genes transcribed in each strain. The location and orientation of the erm cassette are shown by a black arrow. The T. denticola strains are 35405 (wild-type parent), P0760 ( erm insertion at the 5′ end of prcB ), CF417 ( prcB modified to encode a C-terminal His tag; erm insertion replaces the 5′ end of prcA ), CF499 ( prcB modified to encode a C-terminal His tag; erm insertion is upstream of the protease locus), and CF522 (Δ prcB ; promoter region is intact; erm insertion as in CF499).

    Journal: Journal of Bacteriology

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex ▿

    doi: 10.1128/JB.00274-10

    Figure Lengend Snippet: Protease locus in T. denticola parent and prcB mutant strains. The genes of the protease operon, prcB , prcA , and prtP , are shown as gray arrows. The location of the protease operon promoter region is indicated by “P.” The protease mRNA transcript is shown as a thin arrow above the genes transcribed in each strain. The location and orientation of the erm cassette are shown by a black arrow. The T. denticola strains are 35405 (wild-type parent), P0760 ( erm insertion at the 5′ end of prcB ), CF417 ( prcB modified to encode a C-terminal His tag; erm insertion replaces the 5′ end of prcA ), CF499 ( prcB modified to encode a C-terminal His tag; erm insertion is upstream of the protease locus), and CF522 (Δ prcB ; promoter region is intact; erm insertion as in CF499).

    Article Snippet: Counting from the first AUG codon in the ORF, the deduced amino acid sequences are identical for residues 1 to 65, which includes the predicted untranslated region upstream of the GUG translation initiation codon in the annotated T. denticola genome sequence ( ).

    Techniques: Mutagenesis, Modification

    Conservation of prcB in T. denticola . The deduced amino acid sequence of the complete ORF, including the PrcB coding region, is shown for T. denticola strains ATCC 35405, ATCC 33520, and OTK, aligned using Clustal 2.0. The N-terminal valine residue predicted by JCVI (Val 38 ) and the two possible N-terminal methionine residues (Met 1 and Met 7 ) predicted by the present study are shaded gray. The predicted signal peptidase II cleavage site is indicated by a black arrowhead. Amino acid identity throughout all three sequences is indicated by asterisks. Conserved and semiconserved amino acids are shown by colons and dots, respectively.

    Journal: Journal of Bacteriology

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex ▿

    doi: 10.1128/JB.00274-10

    Figure Lengend Snippet: Conservation of prcB in T. denticola . The deduced amino acid sequence of the complete ORF, including the PrcB coding region, is shown for T. denticola strains ATCC 35405, ATCC 33520, and OTK, aligned using Clustal 2.0. The N-terminal valine residue predicted by JCVI (Val 38 ) and the two possible N-terminal methionine residues (Met 1 and Met 7 ) predicted by the present study are shaded gray. The predicted signal peptidase II cleavage site is indicated by a black arrowhead. Amino acid identity throughout all three sequences is indicated by asterisks. Conserved and semiconserved amino acids are shown by colons and dots, respectively.

    Article Snippet: Counting from the first AUG codon in the ORF, the deduced amino acid sequences are identical for residues 1 to 65, which includes the predicted untranslated region upstream of the GUG translation initiation codon in the annotated T. denticola genome sequence ( ).

    Techniques: Sequencing

    Expression and localization of PrcB. PrcB-6×His and PrtP were detected on blots by use of HisProbe reagent and anti-PrtP antibodies, respectively. Triton X-114 extracts (TX-114) were partitioned into aqueous (Aq) and detergent (Det) phases. Molecular mass standards are shown in panels B and C. (A) Expression of PrcB-6×His detected in lysates of E. coli /pCF415 ( E.c. ) and T. denticola strains CF499 and CF417. (B) PrcB-6×His localizes to the detergent phase of a T. denticola CF499 Triton X-114 extract. (C) T. denticola CF499 Triton X-114 extract showing differential phase partitioning of full-length and N-terminally truncated PrcB-6×His. Samples were heated (+) or not heated (−) prior to SDS-PAGE.

    Journal: Journal of Bacteriology

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex ▿

    doi: 10.1128/JB.00274-10

    Figure Lengend Snippet: Expression and localization of PrcB. PrcB-6×His and PrtP were detected on blots by use of HisProbe reagent and anti-PrtP antibodies, respectively. Triton X-114 extracts (TX-114) were partitioned into aqueous (Aq) and detergent (Det) phases. Molecular mass standards are shown in panels B and C. (A) Expression of PrcB-6×His detected in lysates of E. coli /pCF415 ( E.c. ) and T. denticola strains CF499 and CF417. (B) PrcB-6×His localizes to the detergent phase of a T. denticola CF499 Triton X-114 extract. (C) T. denticola CF499 Triton X-114 extract showing differential phase partitioning of full-length and N-terminally truncated PrcB-6×His. Samples were heated (+) or not heated (−) prior to SDS-PAGE.

    Article Snippet: Counting from the first AUG codon in the ORF, the deduced amino acid sequences are identical for residues 1 to 65, which includes the predicted untranslated region upstream of the GUG translation initiation codon in the annotated T. denticola genome sequence ( ).

    Techniques: Expressing, SDS Page